LOCUS       sequence-449160-       11434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBAC pCAG-hCas9-T2A-Unbiased3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11434)
  AUTHORS   Carlson CK, Loveless TB, Milisavljevic M, Kelly PI, Mills JH, Tyo 
            KEJ, Liu CC
  TITLE     A massively parallel in vivo assay of TdT mutants yields variants 
            with altered nucleotide insertion biases.
  JOURNAL   bioRxiv [Preprint]. 2024 Jun 11:2024.06.11.598561. doi: 
            10.1101/2024.06.11.598561.
  PUBMED    38915690
REFERENCE   2  (bases 1 to 11434)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Jun 11:2024.06.11.598561. doi: 
            10.1101/2024.06.11.598561."
COMMENT     Sequence Label: pCKC764
FEATURES             Location/Qualifiers
     source          1..11434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(264..317)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(327..347)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(360..4463)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(360..4463)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(360..4463)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(4497..4516)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          4524..5540
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(4578..4600)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        5816..6119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   6470..6504
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6533..6552)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6534..6552)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(6664..7252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6744..6763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7466..7485)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7477..8292
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             7477..8292
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(7550..7569)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        8332..8350
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   8401..8463
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    8670..8877
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8866..8883
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(8895..9293)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      9290..9299
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9319..9530)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    9599..9716
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    10021..10069
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     10094..10113
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 agccactcca gaacatccga acccgagttg ttctctgcta caatgtgggt gacagaatca
       61 ctgagctcat tttcaaccct gaaccctttc ctgcgggcca gctccatgag gaacgctctg
      121 cgggtggttc ccattttctt ctccaaaatg aagacgacca aatcttgaaa tttgatgtct
      181 tgaggagagg aggccatcaa ggcacccgtc tgccggggtc tcttcttccg agggctcaag
      241 tgggacgctc gtggtggatc catggggccg gggttctcct ccacgtcgcc gcaggtcagc
      301 aggctgcctc tgccctcgcc gctgcccacc ttcctcttct tcttggggtc agccctgctg
      361 tctccaccga gctgagagag gtcgattctt gtttcataga gccccgtaat tgactgatga
      421 atcagtgtgg cgtccaggac ctcctttgta gaggtgtacc gctttctgtc tatggtggtg
      481 tcgaagtact tgaaggctgc aggcgcgccc aagttggtca gagtaaacaa gtggataatg
      541 ttttctgcct gctccctgat gggcttatcc ctgtgcttat tgtaagcaga aagcacctta
      601 tcgaggttag cgtcggcgag gatcactctt ttggagaatt cgcttatttg ctcgatgatc
      661 tcatcaaggt agtgtttgtg ttgttccacg aacagctgct tctgctcatt atcttcggga
      721 gaccctttga gcttttcata gtggctggcc agatacaaga aattaacgta tttagagggc
      781 agtgccagct cgttaccttt ctgcagctcg cccgcactag cgagcattcg tttccggccg
      841 ttttcaagct caaagagaga gtacttggga agcttaatga tgaggtcttt tttgacctct
      901 ttatatcctt tcgcctcgag aaagtcgatg gggttttttt cgaagcttga tcgctccatg
      961 attgtgatgc ccagcagttc cttgacgctt ttgagttttt tagacttccc tttctccact
     1021 ttggccacaa ccagtacact gtaagcgact gtaggagaat cgaatccgcc gtatttcttg
     1081 gggtcccaat cttttttgcg tgcgatcagc ttgtcgctgt tccttttcgg gaggatactt
     1141 tccttggaga agcctccggt ctgtacttcg gtctttttaa cgatgttcac ctgcggcatg
     1201 gacaggacct tccggactgt cgcgaaatcc ctacccttgt cccacacgat ttctcctgtt
     1261 tctccgtttg tttcgataag tggtcgcttc cgaatctctc cattggccag tgtaatctcg
     1321 gtcttgaaaa aattcataat attgctgtaa aagaagtact tagcggtggc cttgcctatt
     1381 tcctgctcag actttgcgat cattttccta acatcgtaca ctttatagtc tccgtaaaca
     1441 aattcagatt caagcttggg atattttttg ataagtgcag tgcctaccac tgcattcagg
     1501 taggcatcat gcgcatggtg gtaattgttg atctctctca ccttataaaa ctgaaagtcc
     1561 tttctgaaat ctgagaccag cttagacttc agagtaataa ctttcacctc tcgaatcagt
     1621 ttgtcatttt catcgtactt ggtgttcatg cgtgaatcga gaatttgggc cacgtgcttg
     1681 gtgatctggc gtgtctcaac aagctgcctt ttgatgaagc cggctttatc caactcagac
     1741 aggccacctc gttcagcctt agtcagatta tcgaacttcc gttgtgtgat cagtttggcg
     1801 ttcagcagct gccgccaata atttttcatt ttcttgacaa cttcttctga ggggacgtta
     1861 tcactcttcc ctctattttt atcggatctt gtcaacactt tattatcaat agaatcatct
     1921 ttgagaaaag actggggcac gatatgatcc acgtcgtagt cggagagccg attgatgtcc
     1981 agttcctgat ccacgtacat gtccctgccg ttctgcaggt agtacaggta gagcttctca
     2041 ttctgaagct gggtgttttc aactgggtgt tccttaagga tttgggaccc cagttctttt
     2101 ataccctctt caatcctctt catcctttcc ctactgttct tctgtccctt ctgggtagtt
     2161 tggttctctc gggccatctc gataacgata ttctcgggct tatgccttcc cattactttg
     2221 acgagttcat ccacgacctt aacggtctgc agtattccct ttttgatagc tgggctacct
     2281 gcaagattag cgatgtgctc gtgaagactg tccccctggc cagaaacttg tgctttctgg
     2341 atgtcctcct taaaggtgag agagtcatca tggatcaact gcatgaagtt ccggttggca
     2401 aatccatcgg acttaagaaa atccaggatt gtctttccac tctgcttgtc tcggatccca
     2461 ttgatcagtt ttcttgacag ccgcccccat cctgtatatc ggcgcctctt gagctgtttc
     2521 atgactttgt cgtcgaagag atgagcgtaa gttttcaagc gttcttcaat catctcccta
     2581 tcttcaaaca acgtaagggt gaggacaatg tcctcaagaa tgtcctcgtt ctcctcattg
     2641 tccaggaagt ccttgtcttt aatgattttc aggagatcgt gatacgttcc cagggatgcg
     2701 ttgaagcgat cctccactcc gctgatttca acagagtcga aacattcaat ctttttgaaa
     2761 tagtcttctt tgagctgttt cacggtaact ttccggttcg tcttgaagag gaggtccacg
     2821 atagctttct tctgctctcc agacaggaat gctggctttc tcatcccttc tgtgacgtat
     2881 ttgaccttgg tgagctcgtt ataaactgtg aagtactcgt acagcagaga gtgtttagga
     2941 agcacctttt cgttaggcag atttttatca aagttagtca tcctttcgat gaaggactgg
     3001 gcagaggccc ccttatccac gacttcctcg aagttccagg gagtgatggt ctcttctgat
     3061 ttgcgagtca tccacgcgaa tctggaattt ccccgggcga gggggcctac atagtagggt
     3121 atccgaaatg tgaggatttt ctcaatcttt tccctgttat ctttcaaaaa ggggtagaaa
     3181 tcctcttgcc gcctgaggat agcgtgcagt tcgcccaggt gaatctggtg ggggatgctt
     3241 ccattgtcga aagtgcgctg tttgcgcaac agatcttctc tgttaagctt taccagcagc
     3301 tcctcggtgc cgtccatttt ttccaagatg ggcttaataa atttgtaaaa ttcctcctgg
     3361 cttgctccgc cgtcaatgta tccggcgtag ccatttttag actgatcgaa gaaaatttcc
     3421 ttgtacttct caggcagttg ctgtctgaca agggccttca gcaaagtcaa gtcttggtgg
     3481 tgctcatcat agcgcttgat catactagcg ctcagcggag ctttggtgat ctccgtgttc
     3541 actcgcagaa tatcactcag cagaatggcg tctgacaggt tctttgccgc caaaaaaagg
     3601 tctgcgtact ggtcgccgat ctgggccagc agattgtcga gatcatcatc gtaggtgtct
     3661 ttgctcagtt gaagcttggc atcttcggcc aggtcgaagt tagatttaaa gttgggggtc
     3721 agcccgagtg acagggcgat aagattacca aacaggccgt tcttcttctc cccagggagc
     3781 tgtgcgatga ggttttcgag ccgccgggat ttggacagcc tagcgctcag gattgctttg
     3841 gcgtcaactc cggatgcgtt gatcgggttc tcttcgaaaa gctgattgta agtctgaacc
     3901 agttggataa agagtttgtc gacatcgctg ttgtctgggt tcaggtcccc ctcgatgagg
     3961 aagtgtcccc gaaatttgat catatgcgcc agcgcgagat agatcaaccg caagtcagcc
     4021 ttatcagtac tgtctacaag cttcttcctc agatgatata tggttgggta cttttcatgg
     4081 tacgccacct cgtccacgat attgccaaag attgggtggc gctcgtgctt tttatcctcc
     4141 tccaccaaaa aggactcctc cagcctatgg aagaaagagt catccacctt agccatctca
     4201 ttactaaaga tctcctgcag gtagcagatc cgattctttc tgcgggtata tctgcgccgt
     4261 gctgttcttt tgagccgcgt ggcttcggcc gtctccccgg agtcgaacag gagggcgcca
     4321 atgaggttct tctttatgct gtggcgatcg gtattgccca gaactttgaa ttttttgctc
     4381 ggcaccttgt actcgtccgt aatgacggcc cagccgacgc tgtttgtgcc gatatcgagc
     4441 ccaatggagt acttcttgtc catagtggcg aattctttgc caaaatgatg agacagcaca
     4501 ataaccagca cgttgcccag gagctgtagg aaaaagaaga aggcatgaac atggttagca
     4561 gaggctctag agccgccggt cacacgccag aagccgaacc ccgccctgcc ccgtcccccc
     4621 cgaaggcagc cgtccccccg cggacagccc cgaggctgga gagggagaag gggacggcgg
     4681 cgcggcgacg cacgaaggcc ctccccgccc atttccttcc tgccggcgcc gcaccgcttc
     4741 gccccgcgcc cgctagaggg ggtgcggcgg cgcctcccag atttcggctc cgcacagatt
     4801 tgggacaaag gaagtccctg cgccctctcg cacgattacc ataaaaggca atggctgcgg
     4861 ctcgccgcgc ctcgacagcc gccggcactc cgggggccgc cgcgcccctc ccccgagccc
     4921 tccccggccc gaggcggccc cgccccgccc ggcaccccca cctgccgcca ccccccgccc
     4981 ggcacggcga gccccgcgcc acgccccgta cggagccccg cacccgaagc cgggccgtgc
     5041 tcagcaactc ggggaggggg gtgcaggggg ggttgcagcc cgaccgacgc gcccacaccc
     5101 cctgctcacc cccccacgca cacaccccgc acgcagcctt tgttcccctc gcagcccccc
     5161 ccgcaccgcg gggcaccgcc cccggccgcg ctcccctcgc gcacactgcg gagcgcacaa
     5221 agccccgcgc cgcgcccgca gcgctcacag ccgccgggca gcgcggagcc gcacgcggcg
     5281 ctccccacgc acacacacac gcacgcaccc cccgagccgc tccccccgca caaagggccc
     5341 tcccggagcc cctcaaggct ttcacgcagc cacagaaaag aaacaagccg tcattaaacc
     5401 aagcgctaat tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac
     5461 gcggtcagtc agagccgggg cgggcggcgc gaggcggcgg cggagcgggg cacggggcga
     5521 aggcagcgcg cagcgactcc cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc
     5581 cgccgcctcg ccataaaagg aaactttcgg agcgcgccgc tctgattggc tgccgccgca
     5641 cctctccgcc tccccccccc cccccccccc cccccccccc cccccccccg cccccccccc
     5701 cccccccccc cccccccgct gcacaaaata attaaaaaat aaataaatac aaaattgggg
     5761 gtggggaggg gggggagatg gggagagtga agcagaacgt ggggctcacc tcgaccatgg
     5821 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     5881 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     5941 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     6001 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     6061 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     6121 ggaactccat atatggtcag tctagatgcg aattcagggc ccatggagat ctgggcccat
     6181 gctagcattc tagttgatat ctataacaag aaaatatata tataataagt tatcacgtaa
     6241 gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag tcacgtaaaa
     6301 gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag tgacacttac
     6361 cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc ctaaatgcac
     6421 agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat
     6481 tttacgcaga ctatctttct agggttaatc tagaatggcg cgccccaatt cgccctatag
     6541 tgagtcgtat taattggcgg ccgcacggtt atccacagaa tcaggggata acgcaggaaa
     6601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7261 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7321 ggtcatgact gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccacgttgtg
     7381 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     7441 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     7501 tcttgctcca ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     7561 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     7621 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     7681 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     7741 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
     7801 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     7861 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc
     7921 ctcgctcagg cgcaatcacg aatgaataat ggtttggttg atgcgagtga ttttgatgac
     7981 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc
     8041 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     8101 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     8161 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     8221 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     8281 gagtttttct aatcagaatt ggttaattgg ttgtaacacg cggccgctcg gaattaaccc
     8341 tcactaaagg gaacaaaagc tggattaatt aagcggccgc atacgcgtat actagattaa
     8401 ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgcgta aaattgacgc
     8461 atgtgtttta tcggtctgta tatcgaggtt tatttattaa tttgaataga tattaagttt
     8521 tattatattt acacttacat actaataata aattcaacaa acaatttatt tatgtttatt
     8581 tatttattaa aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttaac
     8641 tagtatcgat actcatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
     8701 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
     8761 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     8821 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     8881 aggcgtcgct cgagttagcc ctcccacaca taaccagagg gcagcaattc acgaatccca
     8941 actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg atgcagatcg
     9001 agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc atttccgatc
     9061 gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag caccacgagt
     9121 tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat gcggccgtcg
     9181 ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct gttgattgta
     9241 gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc catggtggcg
     9301 gatcccgcgt cacgacacct gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac
     9361 ggcgactact gcacttatat acggttctcc cccaccctcg ggaaaaaggc ggagccagta
     9421 cacgacatca ctttcccagt ttaccccgcg ccaccttctc taggcaccgg ttcaattgcc
     9481 gacccctccc cccaacttct cggggactgt gggcgatgtg cgctctgccc actgacgggc
     9541 accggagcca ctcgagccgg cgccaaagtg gatctctgct gtccctgtaa taaacccgaa
     9601 aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta
     9661 tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt
     9721 ggatgaatac tgccatttgt ctcaagatct atacgtaggt accccataga gcccaccgct
     9781 gtaaggttag tgtgtccagc atgcaacctt tccttcctgg atgagcatcc ctggaatagg
     9841 agagccagcc tgcctcctgc gctggcacag agcccggttc cctagacaac tgcctctcca
     9901 aatctgatgt ccagcgccac ctggtgtcca catcaagcag acacaattaa tagtcaacct
     9961 gttcagggtt cgtaacatct ctgtaaccag cccagccctg ggaccgaacc ccgcgtttat
    10021 gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg
    10081 gttccttccg gtattgtctc cttccgtgtt tcagttagcc tcccccgttt aaactcatta
    10141 ctaaccggta gggatcgaac cctttcacta ggcatttctt tcccacggtt caatataatc
    10201 caatcccaga tgcgcaaaaa tttcttcttc actttctgct ttgaggaata tcctcttggt
    10261 cttgtcatat aaagcatggt tatccagaat catcttccgc tcatgtgtgg catagcgccg
    10321 gaggtctctc tcaaactgcc gggagccagt ccatcccaac agggcaaagg cacgacgctc
    10381 gtaggggcac agaactaaat ccacacggat ggccttccag gtctttcctt cctgccagct
    10441 ggactggtca ctgtccactc tttgacgagg caatttgaaa atcagaaagc acttttgaaa
    10501 atgatcaaaa gcccgaacct tcctgctagg caacctgagc ttttcaaatg ttgactccac
    10561 aaggtcataa tataaaagta atcccttctt ttcccataag ttcatcactt tctgtaaaag
    10621 ttgctcttca tcctctgttg atcctgggct ggtaattaaa aaatctacat catgccccat
    10681 cttcttaccc ctccggaacc ctcctgtcat ggtgacgaaa gcatccggaa gaaatgccca
    10741 gacagcctct ttaaccagca cactgacggc ctctgcttct gccctggtca cacagctgac
    10801 aaggtcttca taatacagaa atcctgcttt ctgcattcgt gtaaatttca ggcttttgtc
    10861 cgaccttact ttactcagag ttctgaaacc catcctgaac cacttctcag aagtcttcag
    10921 ccccactcca aatacagaag taaagagttt gaaggattga tatcgttcat catttaacac
    10981 agctttaact tcagaacttt ctccatcttc aataatctcc tctatgatac ccttcacctt
    11041 ggaccccagg cagggaattc cttctgtgtc cttcatactg atgattgtga atggcagaga
    11101 tttcaataca gaagctgctc tcataaatgt cacacaggag tcttcatttt ctctaaactc
    11161 acagttttca gccagtatat caaaggcatc cgtgaatatc tggttacagt tgtttaaagt
    11221 ggttcttctc tgacacgcat actgggagat cttttgtaca gcaattggtg gagtcttcgg
    11281 ggggcctggg ttggtgctat ctgaatagtc tcttctcaca acaagctggt gttttcctgt
    11341 catttccacc ggtttccctg ctcctatgca ttcgatcagc caggagacat cgaggagctc
    11401 tggttgtgag ctgacttgta ctttctgtgc ttga
//
