
|------------------------------------------------------------------|
| MCPrimers V2.4 Copyright (c) 2005,2006 Stephen G. Lenk           |
| CloningVector  Copyright (c) 2006 Tim Wiggin and Stephen G. Lenk |
| Primer3        Copyright (c) 1996,1997,1998,1999,2000,2001,2004  |
| Whitehead Institute for Biomedical Research. All rights reserved |
|------------------------------------------------------------------|

Date                 = Thu Sep  7 23:48:24 2006
Sequence file        = ppib3inarow.fa
Results file         = 
Primer3 file         = 
Cloning vector       = pet-32a.txt
Clamp flag           = both
Maximum changes      = 3
Search past start    = 0
Search before stop   = 0
Excluded sites       = 
Original sequence:

TTAGATGGTAT


=========
=========  Solution # 1
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = EcoRI (GAATTC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       600   21   57.16   42.86  6.00  0.00 GCGAGAATTCACCATAATCAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 593, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTGAATTCTCG
                                              <<<<<<<<<<<<<<<<<<<<

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <                                                           

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 2
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SacI (GAGCTC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       630   19   58.14   57.89  8.00  0.00 GTGGGAGCTCTCCATCAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 623, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGAGCTCCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                  <<<<<<<<<<<<<<<<<<<                             

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 3
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SacI (GAGCTC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       681   22   59.89   40.91  8.00  1.00 CATTGAGCTCAGCATTCAAAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 674, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTGAGCTCAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
      <<<<<<<<<<<<<<<<<<<<<<                                      

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 4
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SacI (GAGCTC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       561   24   59.83   41.67  6.00  2.00 CTTTAGGGAGCTCTTGTTTTATGC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 554, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX       <<

  540 ATAAAACAAGAGCTCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<<<<<<<<<                                      

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 5
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       638   21   61.94   52.38  7.00  3.00 CATTTCAGGTCGACGATCTCC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 631, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCGTCGACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                        <<<<<<<<<<<<<<<<<<<<<                     

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 6
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       561   23   57.96   39.13  6.00  0.00 CTTTAGGTCGACGTTGTTTTATG
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 554, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX        <

  540 ATAAAACAACGTCGACCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<<<<<<<<<                                      

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 7
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       600   21   59.59   47.62  6.00  2.00 GCGAAAGTCGACCATAATCAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 593, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCGACTTTCG
                                              <<<<<<<<<<<<<<<<<<<<

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <                                                           

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 8
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       582   22   59.64   45.45  6.00  1.00 CACATTGTCGACTTTCTCCAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 575, PAIR ANY COMPL: 7.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAAGTCGACAATGTGATTATGGTCAATTTTCG
                           <<<<<<<<<<<<<<<<<<<<<<                 

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 9
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       553   23   61.87   43.48  6.00  2.00 CGATGTCGACTTATGCTCATTTG
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 546, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX<<<<<<<<<

  540 ATAAGTCGACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<                                              

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 10
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       681   22   59.48   40.91  6.00  0.00 CATTCAGTCGACCATTCAAAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 674, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGGTCGACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
      <<<<<<<<<<<<<<<<<<<<<<                                      

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 11
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       706   24   59.47   37.50  8.00  2.00 GAAATTGGTCGACCTAAAATAAGC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 699, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGGTCGACCAATTTCATCTAAAATTAAT
                             <<<<<<<<<<<<<<<<<<<<<<<<             

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 12
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       604   23   60.01   43.48  6.00  2.00 GTATGCGAAGCTTGACCATAATC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 597, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAAGCTTCG
                                                <<<<<<<<<<<<<<<<<<

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <<<<<                                                       

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 13
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       677   24   57.36   33.33  6.00  3.00 CAGTTAAGCTTTCAAAACAACAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 670, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                            <<<<<<

  660 GTTTTGAAAGCTTAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
      <<<<<<<<<<<<<<<<<<                                          

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 14
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       670   22   60.99   40.91  6.00  0.00 GCATTCAAAGCTTCAACCAAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 663, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGAA
                                                       <<<<<<<<<<<

  660 GCTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
      <<<<<<<<<<<                                                 

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 15
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       619   21   57.41   47.62  6.00  2.00 CCATAAGCTTGACAGGTATGC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 612, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                 <

  600 CATACCTGTCAAGCTTATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <<<<<<<<<<<<<<<<<<<<                                        

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 16
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       628   22   61.02   45.45  6.00  3.00 GGAAAAGCTTCATCAACGTGAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 621, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 3.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGAAGCTTTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
             <<<<<<<<<<<<<<<<<<<<<<                               

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 17
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       556   21   61.10   47.62  8.00  0.00 GGGCAAGCTTGTTTTATGCTC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 549, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX     <<<<

  540 ATAAAACAAGCTTGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<<<<                                           

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 18
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       627   19   57.01   52.63  6.00  2.00 GAAGATCTCGAGCAACGTG
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 620, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGCTCGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
               <<<<<<<<<<<<<<<<<<<                                

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 19
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       582   22   58.84   45.45  6.00  1.00 CACATTCTCGAGTTTCTCCAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 575, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAACTCGAGAATGTGATTATGGTCAATTTTCG
                           <<<<<<<<<<<<<<<<<<<<<<                 

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 20
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       600   21   59.87   47.62  6.00  1.00 GCGAAACTCGAGCATAATCAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 593, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGCTCGAGTTTCG
                                              <<<<<<<<<<<<<<<<<<<<

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <                                                           

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 21
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       683   19   59.18   52.63  6.00  2.00 CCCATTCACTCGAGCATTC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 676, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTCGAGTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
           <<<<<<<<<<<<<<<<<<<                                    

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 22
=========

Start codon at  21
Stop codon  at  528
Left RE site  = BamHI (GGATCC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   60.44   52.38  6.00  3.00 GAGGGGATCCACAATGGTTAC
RIGHT PRIMER       561   24   60.62   41.67  6.00  2.00 CTTTAGCTCGAGGTTGTTTTATGC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 554, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGGATCCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX       <<

  540 ATAAAACAACCTCGAGCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<<<<<<<<<                                      

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 23
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       638   21   61.94   52.38  7.00  3.00 CATTTCAGGTCGACGATCTCC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 631, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCGTCGACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                        <<<<<<<<<<<<<<<<<<<<<                     

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 24
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       561   23   57.96   39.13  6.00  0.00 CTTTAGGTCGACGTTGTTTTATG
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 554, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX        <

  540 ATAAAACAACGTCGACCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<<<<<<<<<                                      

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 25
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       600   21   59.59   47.62  6.00  2.00 GCGAAAGTCGACCATAATCAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 593, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCGACTTTCG
                                              <<<<<<<<<<<<<<<<<<<<

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <                                                           

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 26
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       582   22   59.64   45.45  6.00  1.00 CACATTGTCGACTTTCTCCAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 575, PAIR ANY COMPL: 7.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAAGTCGACAATGTGATTATGGTCAATTTTCG
                           <<<<<<<<<<<<<<<<<<<<<<                 

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 27
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       553   23   61.87   43.48  6.00  2.00 CGATGTCGACTTATGCTCATTTG
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 546, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX<<<<<<<<<

  540 ATAAGTCGACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<                                              

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 28
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       681   22   59.48   40.91  6.00  0.00 CATTCAGTCGACCATTCAAAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 674, PAIR ANY COMPL: 6.00, PAIR 3' COMPL: 3.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGGTCGACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
      <<<<<<<<<<<<<<<<<<<<<<                                      

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 29
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = SalI (GTCGAC)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       706   24   59.47   37.50  8.00  2.00 GAAATTGGTCGACCTAAAATAAGC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 699, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGGTCGACCAATTTCATCTAAAATTAAT
                             <<<<<<<<<<<<<<<<<<<<<<<<             

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 30
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       604   23   60.01   43.48  6.00  2.00 GTATGCGAAGCTTGACCATAATC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 597, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAAGCTTCG
                                                <<<<<<<<<<<<<<<<<<

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <<<<<                                                       

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 31
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       677   24   57.36   33.33  6.00  3.00 CAGTTAAGCTTTCAAAACAACAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 670, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                            <<<<<<

  660 GTTTTGAAAGCTTAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
      <<<<<<<<<<<<<<<<<<                                          

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 32
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       670   22   60.99   40.91  6.00  0.00 GCATTCAAAGCTTCAACCAAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 663, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGAA
                                                       <<<<<<<<<<<

  660 GCTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
      <<<<<<<<<<<                                                 

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 33
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       619   21   57.41   47.62  6.00  2.00 CCATAAGCTTGACAGGTATGC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 612, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                 <

  600 CATACCTGTCAAGCTTATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <<<<<<<<<<<<<<<<<<<<                                        

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 34
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       628   22   61.02   45.45  6.00  3.00 GGAAAAGCTTCATCAACGTGAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 621, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 3.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGAAGCTTTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
             <<<<<<<<<<<<<<<<<<<<<<                               

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 35
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = HindIII (AAGCTT)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       556   21   61.10   47.62  8.00  0.00 GGGCAAGCTTGTTTTATGCTC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 549, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 3.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX     <<<<

  540 ATAAAACAAGCTTGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<<<<                                           

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 36
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       627   19   57.01   52.63  6.00  2.00 GAAGATCTCGAGCAACGTG
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 620, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGCTCGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
               <<<<<<<<<<<<<<<<<<<                                

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 37
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       582   22   58.84   45.45  6.00  1.00 CACATTCTCGAGTTTCTCCAAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 575, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAACTCGAGAATGTGATTATGGTCAATTTTCG
                           <<<<<<<<<<<<<<<<<<<<<<                 

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 38
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       600   21   59.87   47.62  6.00  1.00 GCGAAACTCGAGCATAATCAC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 593, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 2.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGCTCGAGTTTCG
                                              <<<<<<<<<<<<<<<<<<<<

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
      <                                                           

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 39
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       683   19   59.18   52.63  6.00  2.00 CCCATTCACTCGAGCATTC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 676, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 1.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX         

  540 ATAAAACAACATCGCCCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
                                                                  

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTCGAGTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
           <<<<<<<<<<<<<<<<<<<                                    

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


=========
=========  Solution # 40
=========

Start codon at  21
Stop codon  at  528
Left RE site  = SacI (GAGCTC)
Right RE site = AvaI-XhoI (CTCGAG)

Primer3 analysis of PCR primers designed by MCPrimers:

No mispriming library specified
Using 0-based sequence positions
OLIGO            start  len      tm     gc%   any    3' seq
LEFT PRIMER          8   21   58.66   52.38  6.00  3.00 GAGGGAGCTCACAATGGTTAC
RIGHT PRIMER       561   24   60.62   41.67  6.00  2.00 CTTTAGCTCGAGGTTGTTTTATGC
SEQUENCE SIZE: 731
INCLUDED REGION SIZE: 731

PRODUCT SIZE: 554, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
EXCLUDED REGIONS (start, len)*: 39,492

    0 ATTTTTTAGAGGGAGCTCACAATGGTTACATTACACACAAACTTTGGCGACATTAAAATT
              >>>>>>>>>>>>>>>>>>>>>          XXXXXXXXXXXXXXXXXXXXX

   60 AAATTGGATTTTGATAAAGCTCCAGTTACAGCAGAAAACTTCTTAAATTACTGCAAAGAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  120 GGTTTTTATAATAACACAATTTTTCATCGTGTTATTGATGGCTTTATGATTCAAGGTGGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  180 GGTATGGAAAGCGGTATGCGCGAAAAAGCAACTAAAGCCCCAATCCAAAATGAAGCGAAC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  240 AATCGTTTAAGTAACAAACGTGGAACCATTGCAATGGCTCGCACTTCAGATCCACATTCA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  300 GCGACAGCACAATTTTTCATTAACGTGGCAGACAATGATTTCTTAAATTATCGTTCAAAA
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  360 GAAATGTTTGGTCGCGAAGTCGTTCAAGAATGGGGCTATGCAGTATTTGGCGAAGTGGTT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  420 GAAGGTATGGATGTTGTGGATAAAATCAAAAAAGTGAAAACAGGCAACAAAGGCTTTCAT
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

  480 CAAGATGTTCCAACTGAAGATGTAGTGATTACATCGGTTTCTATTGAGTAACAAATGAGC
      XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX       <<

  540 ATAAAACAACCTCGAGCTAAAGTTGGAGAAATTCTAGAATGTGATTATGGTCAATTTTCG
      <<<<<<<<<<<<<<<<<<<<<<                                      

  600 CATACCTGTCACGTTGATGGAGATCTTCCACCTGAAATGGTTAAAAAACGTTTGGTTGTT
                                                                  

  660 GTTTTGAATGCTAAACTGAATGGGCTTATTTTAGTTGCACCAATTTCATCTAAAATTAAT
                                                                  

  720 TTAGATGGTAT
                 

KEYS (in order of precedence):
XXXXXX excluded region
>>>>>> left primer
<<<<<< right primer


Statistics
         con   too    in    in          no    tm    tm  high  high        high      
         sid  many   tar  excl   bad    GC   too   too   any    3'  poly   end      
        ered    Ns   get   reg   GC% clamp   low  high compl compl     X  stab    ok
Left       1     0     0     0     0     0     0     0     0     0     0     0     1
Right      1     0     0     0     0     0     0     0     0     0     0     0     1
Pair Stats:
considered 1, ok 1
primer3 release 1.0


