pTYB1: Difference between revisions

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==At a Glance {{HelpIcon|Help:Plasmid}}==
==At a Glance ==
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[[Image:PTYB1.jpg|thumb|500px|right|pTYB1, Image provided by [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/maps/pTYB1_map.pdf NEB].]]<protect><!--box uid=63ce1649aab8b3180235b0af7618f667.0.T46e17b9aab956-->
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{|{{Prettytable}} width='30%' align=left
{| {{Prettytable}} width='30%' align=left id="T46e17b9aab956"  class=" tableEdit PlasmidTable" 
 
|-
|-
!{{table heading style}} |Type
!align=left {{table heading style}} |Type
||
Cloning & Expression
|-
|-
!{{table heading style}} |Origin of Replication
!align=left {{table heading style}} |Origin of Replication
||
pMB1 <br />
nM13
|-
|-
!{{table heading style}} |Copy #
!align=left {{table heading style}} |Copy #
||
~20 per cell
|-
|-
!{{table heading style}} |Markers
!align=left {{table heading style}} |Markers
||
Ampicillin
|-
|-
!{{table heading style}} |Link to Sequence
!align=left {{table heading style}} |Link to Sequence
||
[http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/sequences/ptyb1.txt NEB]
|-
|-
!{{table heading style}} |Comments
!align=left {{table heading style}} |Features
||
rop <br />
lacI <ref name='PMID:1902522'/> <br />
T7 Promoter <ref name='PMID:1902522'/> <br />
expression ORF: <br />
MSC <br />
Sce VMA intein <ref name='PMID:9224900'/> <br />
CBD <ref name='PMID:9224900'/><ref name='PMID:8045877'/> <br />
|-
|-
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!align=left {{table heading style}} |Accession Number
||
 
 
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==History {{HelpIcon|Help:Plasmid}}==
==Notes==
{{PlasmidHistory}}
The pTYB1 is an E. coli vector created for fusion protein expression and purification. A target gene can be inserted into the the ORF by use of the MCS upstream of the Sce VMA intein. The NdeI site in the MCS has the initiator sequence ATG that enables translation of the ORF. The chitin-binding domain(CBD) downstream of the intein results in a target protein-intein with C-terminal chitin beads that facilitate protein purification. Release of the protein is induced by the thiol cleavage activity of the intein.


==Sources {{HelpIcon|Help:Plasmid}}==
==History ==
{{PlasmidSources}}
pTYB1 is derived from E. coli plamid pBR322, its chitin-binding domain(CBD) is isolated from B. circulans, and the VMA intein from S. cerevisiae.
 
==Sources ==
[http://www.neb.com/nebecomm/products/productN6701.asp New England Biolabs]


==References==
==References==
{{BacktoTop}}
{{RefHelp}}
{{RefHelp}}
<references/>
<references />
 
==Sequence {{HelpIcon|Help:Plasmid}}==
{{PlasmidSeq}}


==Sequence ==
      1  AACTACGTCA GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT
      51  GTTTATTTTT CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA
    101  CCCTGATAAA TGCTTCAATA ATATTGAAAA AGGAAGAGTA TGAGTATTCA
    151  ACATTTCCGT GTCGCCCTTA TTCCCTTTTT TGCGGCATTT TGCCTTCCTG
    201  TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC TGAAGATCAG
    251  TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT
    301  CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA
    351  AAGTTCTGCT ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG
    401  CAACTCGGTC GCCGCATACA CTATTCTCAG AATGACTTGG TTGAGTACTC
    451  ACCAGTCACA GAAAAGCATC TTACGGATGG CATGACAGTA AGAGAATTAT
    501  GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA CTTACTTCTG
    551  ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG
    601  GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA
    651  TACCAAACGA CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG
    701  TTGCGCAAAC TATTAACTGG CGAACTACTT ACTCTAGCTT CCCGGCAACA
    751  ATTAATAGAC TGGATGGAGG CGGATAAAGT TGCAGGACCA CTTCTGCGCT
    801  CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG AGCCGGTGAG
    851  CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC
    901  CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC
    951  GAAATAGACA GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA
    1001  CTGTCAGACC AAGTTTACTC ATATATACTT TAGATTGATT TACCCCGGTT
    1051  GATAATCAGA AAAGCCCCAA AAACAGGAAG ATTGTATAAG CAAATATTTA
    1101  AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT TTTTGTTAAA
    1151  TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA
    1201  TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA
    1251  GAGTCCACTA TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG
    1301  TCTATCAGGG CGATGGCCCA CTACGTGAAC CATCACCCAA ATCAAGTTTT
    1351  TTGGGGTCGA GGTGCCGTAA AGCACTAAAT CGGAACCCTA AAGGGAGCCC
    1401  CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG AGAAAGGAAG
    1451  GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC
    1501  ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG
    1551  CGCGTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA
    1601  ATCCCTTAAC GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA
    1651  GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT
    1701  TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA
    1751  GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT
    1801  ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA
    1851  ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG
    1901  GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG
    1951  ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA
    2001  CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG
    2051  CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG
    2101  GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC
    2151  CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC
    2201  TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG
    2251  GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC
    2301  CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC TGTGGATAAC
    2351  CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC
    2401  CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA
    2451  ATCTGCTCTG ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA
    2501  CGTGACTGGG TCATGGCTGC GCCCCGACAC CCGCCAACAC CCGCTGACGC
    2551  GCCCTGACGG GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA
    2601  CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC ATCACCGAAA
    2651  CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC
    2701  ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA
    2751  GCGTTAATGT CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT
    2801  TCCTGTTTGG TCACTGATGC CTCCGTGTAA GGGGGATTTC TGTTCATGGG
    2851  GGTAATGATA CCGATGAAAC GAGAGAGGAT GCTCACGATA CGGGTTACTG
    2901  ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA ACAACTGGCG
    2951  GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC
    3001  GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA
    3051  TGGCCTGCTT CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT
    3101  TGAGCGAGGG CGTGCAAGAT TCCGAATACC GCAAGCGACA GGCCGATCAT
    3151  CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC GAAAATGACC CAGAGCGCTG
    3201  CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT CATAAGTGCG
    3251  GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA
    3301  GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC
    3351  TTACATTAAT TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG
    3401  TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT
    3451  GCGTATTGGG CGCCAGGGTG GTTTTTCTTT TCACCAGTGA GACGGGCAAC
    3501  AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA GCAAGCGGTC
    3551  CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG
    3601  GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG
    3651  ATATCCGCAC CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC
    3701  CAGCGCCATC TGATCGTTGG CAACCAGCAT CGCAGTGGGA ACGATGCCCT
    3751  CATTCAGCAT TTGCATGGTT TGTTGAAAAC CGGACATGGC ACTCCAGTCG
    3801  CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA GATATTTATG
    3851  CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA
    3901  ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT
    3951  CGCGTACCGT CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC
    4001  AGAGACATCA AGAAATAACG CCGGAACATT AGTGCAGGCA GCTTCCACAG
    4051  CAATGGCATC CTGGTCATCC AGCGGATAGT TAATGATCAG CCCACTGACG
    4101  CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT CGACGCCGCT
    4151  TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG
    4201  ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG
    4251  GTGGCAACGC CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC
    4301  GCGGTTGGGA ATGTAATTCA GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC
    4351  GCGTTTTCGC AGAAACGTGG CTGGCCTGGT TCACCACGCG GGAAACGGTC
    4401  TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG TTACTGGTTT
    4451  CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC
    4501  CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC
    4551  CTTATGCGAC TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT
    4601  TGAGCACCGC CGCCGCAAGG AATGGTGCAT GCCGGCATGC CGCCCTTTCG
    4651  TCTTCAAGAA TTAATTCCCA ATTCCCCAGG CATCAAATAA AACGAAAGGC
    4701  TCAGTCGAAA GACTGGGCCT TTCGTTTTAT CTGTTGTTTG TCGGTGAACG
    4751  CTCTCCTGAG TAGGACAAAT CCGCCGGGAG CGGATTTGAA CGTTGCGAAG
    4801  CAACGGCCCG GAGGGTGGCG GGCAGGACGC CCGCCATAAA CTGCCAGGAA
    4851  TTAATTCCCC AGGCATCAAA TAAAACGAAA GGCTCAGTCG AAAGACTGGG
    4901  CCTTTCGTTT TATCTGTTGT TTGTCGGTGA ACGCTCTCCT GAGTAGGACA
    4951  AATCCGCCGG GAGCGGATTT GAACGTTGCG AAGCAACGGC CCGGAGGGTG
    5001  GCGGGCAGGA CGCCCGCCAT AAACTGCCAG GAATTAATTC CCCAGGCATC
    5051  AAATAAAACG AAAGGCTCAG TCGAAAGACT GGGCCTTTCG TTTTATCTGT
    5101  TGTTTGTCGG TGAACGCTCT CCTGAGTAGG ACAAATCCGC CGGGAGCGGA
    5151  TTTGAACGTT GCGAAGCAAC GGCCCGGAGG GTGGCGGGCA GGACGCCCGC
    5201  CATAAACTGC CAGGAATTAA TTCCCCAGGC ATCAAATAAA ACGAAAGGCT
    5251  CAGTCGAAAG ACTGGGCCTT TCGTTTTATC TGTTGTTTGT CGGTGAACGC
    5301  TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC GTTGCGAAGC
    5351  AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGAAT
    5401  TAATTCCCCA GGCATCAAAT AAAACGAAAG GCTCAGTCGA AAGACTGGGC
    5451  CTTTCGTTTT ATCTGTTGTT TGTCGGTGAA CGCTCTCCTG AGTAGGACAA
    5501  ATCCGCCGGG AGCGGATTTG AACGTTGCGA AGCAACGGCC CGGAGGGTGG
    5551  CGGGCAGGAC GCCCGCCATA AACTGCCAGG AATTGGGGAT CGGAATTAAT
    5601  TCCCGGTTTA AACCGGGGAT CTCGATCCCG CGAAATTAAT ACGACTCACT
    5651  ATAGGGGAAT TGTGAGCGGA TAACAATTCC CCTCTAGAAA TAATTTTGTT
    5701  TAACTTTAAG AAGGAGATAT ACATATGGCT AGCTCGCGAG TCGACGGCGG
    5751  CCGCGAATTC CTCGAGGGCT CTTCCTGCTT TGCCAAGGGT ACCAATGTTT
    5801  TAATGGCGGA TGGGTCTATT GAATGTATTG AAAACATTGA GGTTGGTAAT
    5851  AAGGTCATGG GTAAAGATGG CAGACCTCGT GAGGTAATTA AATTGCCCAG
    5901  AGGAAGAGAA ACTATGTACA GCGTCGTGCA GAAAAGTCAG CACAGAGCCC
    5951  ACAAAAGTGA CTCAAGTCGT GAAGTGCCAG AATTACTCAA GTTTACGTGT
    6001  AATGCGACCC ATGAGTTGGT TGTTAGAACA CCTCGTAGTG TCCGCCGTTT
    6051  GTCTCGTACC ATTAAGGGTG TCGAATATTT TGAAGTTATT ACTTTTGAGA
    6101  TGGGCCAAAA GAAAGCCCCC GACGGTAGAA TTGTTGAGCT TGTCAAGGAA
    6151  GTTTCAAAGA GCTACCCAAT ATCTGAGGGG CCTGAGAGAG CCAACGAATT
    6201  AGTAGAATCC TATAGAAAGG CTTCAAATAA AGCTTATTTT GAGTGGACTA
    6251  TTGAGGCCAG AGATCTTTCT CTGTTGGGTT CCCATGTTCG TAAAGCTACC
    6301  TACCAGACTT ACGCTCCAAT TCTTTATGAG AATGACCACT TTTTCGACTA
    6351  CATGCAAAAA AGTAAGTTTC ATCTCACCAT TGAAGGTCCA AAAGTACTTG
    6401  CTTATTTACT TGGTTTATGG ATTGGTGATG GATTGTCTGA CAGGGCAACT
    6451  TTTTCGGTTG ATTCCAGAGA TACTTCTTTG ATGGAACGTG TTACTGAATA
    6501  TGCTGAAAAG TTGAATTTGT GCGCCGAGTA TAAGGACAGA AAAGAACCAC
    6551  AAGTTGCCAA AACTGTTAAT TTGTACTCTA AAGTTGTCAG AGGTAATGGT
    6601  ATTCGCAATA ATCTTAATAC TGAGAATCCA TTATGGGACG CTATTGTTGG
    6651  CTTAGGATTC TTGAAGGACG GTGTCAAAAA TATTCCTTCT TTCTTGTCTA
    6701  CGGACAATAT CGGTACTCGT GAAACATTTC TTGCTGGTCT AATTGATTCT
    6751  GATGGCTATG TTACTGATGA GCATGGTATT AAAGCAACAA TAAAGACAAT
    6801  TCATACTTCT GTCAGAGATG GTTTGGTTTC CCTTGCTCGT TCTTTAGGCT
    6851  TAGTAGTCTC GGTTAACGCA GAACCTGCTA AGGTTGACAT GAATGTCACC
    6901  AAACATAAAA TTAGTTATGC TATTTATATG TCTGGTGGAG ATGTTTTGCT
    6951  TAACGTTCTT TCGAAGTGTG CCGGCTCTAA AAAATTCAGG CCTGCTCCCG
    7001  CCGCTGCTTT TGCACGTGAG TGCCGCGGAT TTTATTTCGA GTTACAAGAA
    7051  TTGAAGGAAG ACGATTATTA TGGGATTACT TTATCTGATG ATTCTGATCA
    7101  TCAGTTTTTG CTTGGATCCC AGGTTGTCGT CCATGCATGC GGTGGCCTGA
    7151  CCGGTCTGAA CTCAGGCCTC ACGACAAATC CTGGTGTATC CGCTTGGCAG
    7201  GTCAACACAG CTTATACTGC GGGACAATTG GTCACATATA ACGGCAAGAC
    7251  GTATAAATGT TTGCAGCCCC ACACCTCCTT GGCAGGATGG GAACCATCCA
    7301  ACGTTCCTGC CTTGTGGCAG CTTCAATGAC TGCAGGAAGG GGATCCGGCT
    7351  GCTAACAAAG CCCGAAAGGA AGCTGAGTTG GCTGCTGCCA CCGCTGAGCA
    7401  ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG AGGGGTTTTT
    7451  TGCTGAAAGG AGGAACTATA TCCGGAT






[[Category:Plasmids]]
[[Category:Plasmids]][[Category:Cloning Plasmids]][[Category:Expression Plasmids]][[Category:NEB Plasmids]][[Category:Ampicillin Marker Plasmids]]
[[Category:lacI]][[Category:T7 Promoter]][[Category:MCS]]
__NOTOC__
__NOTOC__

Latest revision as of 21:37, 22 August 2011

At a Glance

pTYB1, Image provided by NEB.


Type

Cloning & Expression

Origin of Replication

pMB1
nM13

Copy #

~20 per cell

Markers

Ampicillin

Link to Sequence

NEB

Features

rop
lacI [1]
T7 Promoter [1]
expression ORF:
MSC
Sce VMA intein [2]
CBD [2][3]

Accession Number


edit table



Notes

The pTYB1 is an E. coli vector created for fusion protein expression and purification. A target gene can be inserted into the the ORF by use of the MCS upstream of the Sce VMA intein. The NdeI site in the MCS has the initiator sequence ATG that enables translation of the ORF. The chitin-binding domain(CBD) downstream of the intein results in a target protein-intein with C-terminal chitin beads that facilitate protein purification. Release of the protein is induced by the thiol cleavage activity of the intein.

History

pTYB1 is derived from E. coli plamid pBR322, its chitin-binding domain(CBD) is isolated from B. circulans, and the VMA intein from S. cerevisiae.

Sources

New England Biolabs

References

[back to top]


See Help:References for how to manage references in EcoliWiki.

  1. ↑ 1.0 1.1 Dubendorff, JW & Studier, FW (1991) Controlling basal expression in an inducible T7 expression system by blocking the target T7 promoter with lac repressor. J. Mol. Biol. 219 45-59 PubMed
  2. ↑ 2.0 2.1 Chong, S et al. (1997) Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element. Gene 192 271-81 PubMed
  3. ↑ Watanabe, T et al. (1994) The roles of the C-terminal domain and type III domains of chitinase A1 from Bacillus circulans WL-12 in chitin degradation. J. Bacteriol. 176 4465-72 PubMed

Sequence

      1  AACTACGTCA GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT 
     51  GTTTATTTTT CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA 
    101  CCCTGATAAA TGCTTCAATA ATATTGAAAA AGGAAGAGTA TGAGTATTCA 
    151  ACATTTCCGT GTCGCCCTTA TTCCCTTTTT TGCGGCATTT TGCCTTCCTG 
    201  TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC TGAAGATCAG 
    251  TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT 
    301  CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA 
    351  AAGTTCTGCT ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG 
    401  CAACTCGGTC GCCGCATACA CTATTCTCAG AATGACTTGG TTGAGTACTC 
    451  ACCAGTCACA GAAAAGCATC TTACGGATGG CATGACAGTA AGAGAATTAT 
    501  GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA CTTACTTCTG 
    551  ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG 
    601  GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA 
    651  TACCAAACGA CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG 
    701  TTGCGCAAAC TATTAACTGG CGAACTACTT ACTCTAGCTT CCCGGCAACA 
    751  ATTAATAGAC TGGATGGAGG CGGATAAAGT TGCAGGACCA CTTCTGCGCT 
    801  CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG AGCCGGTGAG 
    851  CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC 
    901  CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC 
    951  GAAATAGACA GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA 
   1001  CTGTCAGACC AAGTTTACTC ATATATACTT TAGATTGATT TACCCCGGTT 
   1051  GATAATCAGA AAAGCCCCAA AAACAGGAAG ATTGTATAAG CAAATATTTA 
   1101  AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT TTTTGTTAAA 
   1151  TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA 
   1201  TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA 
   1251  GAGTCCACTA TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG 
   1301  TCTATCAGGG CGATGGCCCA CTACGTGAAC CATCACCCAA ATCAAGTTTT 
   1351  TTGGGGTCGA GGTGCCGTAA AGCACTAAAT CGGAACCCTA AAGGGAGCCC 
   1401  CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG AGAAAGGAAG 
   1451  GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC 
   1501  ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG 
   1551  CGCGTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA 
   1601  ATCCCTTAAC GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA 
   1651  GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT 
   1701  TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA 
   1751  GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT 
   1801  ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA 
   1851  ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG 
   1901  GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG 
   1951  ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA 
   2001  CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG 
   2051  CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG 
   2101  GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC 
   2151  CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC 
   2201  TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG 
   2251  GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC 
   2301  CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC TGTGGATAAC 
   2351  CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC 
   2401  CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA 
   2451  ATCTGCTCTG ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA 
   2501  CGTGACTGGG TCATGGCTGC GCCCCGACAC CCGCCAACAC CCGCTGACGC 
   2551  GCCCTGACGG GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA 
   2601  CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC ATCACCGAAA 
   2651  CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC 
   2701  ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA 
   2751  GCGTTAATGT CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT 
   2801  TCCTGTTTGG TCACTGATGC CTCCGTGTAA GGGGGATTTC TGTTCATGGG 
   2851  GGTAATGATA CCGATGAAAC GAGAGAGGAT GCTCACGATA CGGGTTACTG 
   2901  ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA ACAACTGGCG 
   2951  GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC 
   3001  GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA 
   3051  TGGCCTGCTT CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT 
   3101  TGAGCGAGGG CGTGCAAGAT TCCGAATACC GCAAGCGACA GGCCGATCAT 
   3151  CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC GAAAATGACC CAGAGCGCTG 
   3201  CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT CATAAGTGCG 
   3251  GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA 
   3301  GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC 
   3351  TTACATTAAT TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG 
   3401  TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT 
   3451  GCGTATTGGG CGCCAGGGTG GTTTTTCTTT TCACCAGTGA GACGGGCAAC 
   3501  AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA GCAAGCGGTC 
   3551  CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG 
   3601  GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG 
   3651  ATATCCGCAC CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC 
   3701  CAGCGCCATC TGATCGTTGG CAACCAGCAT CGCAGTGGGA ACGATGCCCT 
   3751  CATTCAGCAT TTGCATGGTT TGTTGAAAAC CGGACATGGC ACTCCAGTCG 
   3801  CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA GATATTTATG 
   3851  CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA 
   3901  ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT 
   3951  CGCGTACCGT CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC 
   4001  AGAGACATCA AGAAATAACG CCGGAACATT AGTGCAGGCA GCTTCCACAG 
   4051  CAATGGCATC CTGGTCATCC AGCGGATAGT TAATGATCAG CCCACTGACG 
   4101  CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT CGACGCCGCT 
   4151  TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG 
   4201  ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG 
   4251  GTGGCAACGC CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC 
   4301  GCGGTTGGGA ATGTAATTCA GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC 
   4351  GCGTTTTCGC AGAAACGTGG CTGGCCTGGT TCACCACGCG GGAAACGGTC 
   4401  TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG TTACTGGTTT 
   4451  CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC 
   4501  CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC 
   4551  CTTATGCGAC TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT 
   4601  TGAGCACCGC CGCCGCAAGG AATGGTGCAT GCCGGCATGC CGCCCTTTCG 
   4651  TCTTCAAGAA TTAATTCCCA ATTCCCCAGG CATCAAATAA AACGAAAGGC 
   4701  TCAGTCGAAA GACTGGGCCT TTCGTTTTAT CTGTTGTTTG TCGGTGAACG 
   4751  CTCTCCTGAG TAGGACAAAT CCGCCGGGAG CGGATTTGAA CGTTGCGAAG 
   4801  CAACGGCCCG GAGGGTGGCG GGCAGGACGC CCGCCATAAA CTGCCAGGAA 
   4851  TTAATTCCCC AGGCATCAAA TAAAACGAAA GGCTCAGTCG AAAGACTGGG 
   4901  CCTTTCGTTT TATCTGTTGT TTGTCGGTGA ACGCTCTCCT GAGTAGGACA 
   4951  AATCCGCCGG GAGCGGATTT GAACGTTGCG AAGCAACGGC CCGGAGGGTG 
   5001  GCGGGCAGGA CGCCCGCCAT AAACTGCCAG GAATTAATTC CCCAGGCATC 
   5051  AAATAAAACG AAAGGCTCAG TCGAAAGACT GGGCCTTTCG TTTTATCTGT 
   5101  TGTTTGTCGG TGAACGCTCT CCTGAGTAGG ACAAATCCGC CGGGAGCGGA 
   5151  TTTGAACGTT GCGAAGCAAC GGCCCGGAGG GTGGCGGGCA GGACGCCCGC 
   5201  CATAAACTGC CAGGAATTAA TTCCCCAGGC ATCAAATAAA ACGAAAGGCT 
   5251  CAGTCGAAAG ACTGGGCCTT TCGTTTTATC TGTTGTTTGT CGGTGAACGC 
   5301  TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC GTTGCGAAGC 
   5351  AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGAAT 
   5401  TAATTCCCCA GGCATCAAAT AAAACGAAAG GCTCAGTCGA AAGACTGGGC 
   5451  CTTTCGTTTT ATCTGTTGTT TGTCGGTGAA CGCTCTCCTG AGTAGGACAA 
   5501  ATCCGCCGGG AGCGGATTTG AACGTTGCGA AGCAACGGCC CGGAGGGTGG 
   5551  CGGGCAGGAC GCCCGCCATA AACTGCCAGG AATTGGGGAT CGGAATTAAT 
   5601  TCCCGGTTTA AACCGGGGAT CTCGATCCCG CGAAATTAAT ACGACTCACT 
   5651  ATAGGGGAAT TGTGAGCGGA TAACAATTCC CCTCTAGAAA TAATTTTGTT 
   5701  TAACTTTAAG AAGGAGATAT ACATATGGCT AGCTCGCGAG TCGACGGCGG 
   5751  CCGCGAATTC CTCGAGGGCT CTTCCTGCTT TGCCAAGGGT ACCAATGTTT 
   5801  TAATGGCGGA TGGGTCTATT GAATGTATTG AAAACATTGA GGTTGGTAAT 
   5851  AAGGTCATGG GTAAAGATGG CAGACCTCGT GAGGTAATTA AATTGCCCAG 
   5901  AGGAAGAGAA ACTATGTACA GCGTCGTGCA GAAAAGTCAG CACAGAGCCC 
   5951  ACAAAAGTGA CTCAAGTCGT GAAGTGCCAG AATTACTCAA GTTTACGTGT 
   6001  AATGCGACCC ATGAGTTGGT TGTTAGAACA CCTCGTAGTG TCCGCCGTTT 
   6051  GTCTCGTACC ATTAAGGGTG TCGAATATTT TGAAGTTATT ACTTTTGAGA 
   6101  TGGGCCAAAA GAAAGCCCCC GACGGTAGAA TTGTTGAGCT TGTCAAGGAA 
   6151  GTTTCAAAGA GCTACCCAAT ATCTGAGGGG CCTGAGAGAG CCAACGAATT 
   6201  AGTAGAATCC TATAGAAAGG CTTCAAATAA AGCTTATTTT GAGTGGACTA 
   6251  TTGAGGCCAG AGATCTTTCT CTGTTGGGTT CCCATGTTCG TAAAGCTACC 
   6301  TACCAGACTT ACGCTCCAAT TCTTTATGAG AATGACCACT TTTTCGACTA 
   6351  CATGCAAAAA AGTAAGTTTC ATCTCACCAT TGAAGGTCCA AAAGTACTTG 
   6401  CTTATTTACT TGGTTTATGG ATTGGTGATG GATTGTCTGA CAGGGCAACT 
   6451  TTTTCGGTTG ATTCCAGAGA TACTTCTTTG ATGGAACGTG TTACTGAATA 
   6501  TGCTGAAAAG TTGAATTTGT GCGCCGAGTA TAAGGACAGA AAAGAACCAC 
   6551  AAGTTGCCAA AACTGTTAAT TTGTACTCTA AAGTTGTCAG AGGTAATGGT 
   6601  ATTCGCAATA ATCTTAATAC TGAGAATCCA TTATGGGACG CTATTGTTGG 
   6651  CTTAGGATTC TTGAAGGACG GTGTCAAAAA TATTCCTTCT TTCTTGTCTA 
   6701  CGGACAATAT CGGTACTCGT GAAACATTTC TTGCTGGTCT AATTGATTCT 
   6751  GATGGCTATG TTACTGATGA GCATGGTATT AAAGCAACAA TAAAGACAAT 
   6801  TCATACTTCT GTCAGAGATG GTTTGGTTTC CCTTGCTCGT TCTTTAGGCT 
   6851  TAGTAGTCTC GGTTAACGCA GAACCTGCTA AGGTTGACAT GAATGTCACC 
   6901  AAACATAAAA TTAGTTATGC TATTTATATG TCTGGTGGAG ATGTTTTGCT 
   6951  TAACGTTCTT TCGAAGTGTG CCGGCTCTAA AAAATTCAGG CCTGCTCCCG 
   7001  CCGCTGCTTT TGCACGTGAG TGCCGCGGAT TTTATTTCGA GTTACAAGAA 
   7051  TTGAAGGAAG ACGATTATTA TGGGATTACT TTATCTGATG ATTCTGATCA 
   7101  TCAGTTTTTG CTTGGATCCC AGGTTGTCGT CCATGCATGC GGTGGCCTGA 
   7151  CCGGTCTGAA CTCAGGCCTC ACGACAAATC CTGGTGTATC CGCTTGGCAG 
   7201  GTCAACACAG CTTATACTGC GGGACAATTG GTCACATATA ACGGCAAGAC 
   7251  GTATAAATGT TTGCAGCCCC ACACCTCCTT GGCAGGATGG GAACCATCCA 
   7301  ACGTTCCTGC CTTGTGGCAG CTTCAATGAC TGCAGGAAGG GGATCCGGCT 
   7351  GCTAACAAAG CCCGAAAGGA AGCTGAGTTG GCTGCTGCCA CCGCTGAGCA 
   7401  ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG AGGGGTTTTT 
   7451  TGCTGAAAGG AGGAACTATA TCCGGAT
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