LOCUS       sequence-449161-       10338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBAC 7xTCF-LEF hCas9-T2A-A/T-biased2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10338)
  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 10338)
  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: pCKC775
FEATURES             Location/Qualifiers
     source          1..10338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             272..292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             302..355
                     /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"
     primer_bind     complement(1940..1959)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1984..2032
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    2337..2454
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2523..2734
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2760..3473
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3488..3505)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3494..3701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3908..3970
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(4021..4039)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(4079..4894)
                     /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             complement(4079..4894)
                     /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     4802..4821
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4886..4905
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5119..5707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5608..5627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5819..5838
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5819..5837
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5867..5901
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        6401..6432
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      6488..6497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6494..10338,1..259)
                     /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             join(6494..10338,1..259)
                     /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             join(6494..10338,1..259)
                     /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"
ORIGIN
        1 cgccgacgct aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat
       61 cagggagcag gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc
      121 agccttcaag tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt
      181 cctggacgcc acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct
      241 ctctcagctc ggtggagaca gcagggctga ccccaagaag aagaggaagg tgggcagcgg
      301 cgagggcaga ggcagcctgc tgacctgcgg cgacgtggag gagaaccccg gccccatgga
      361 tccaccacga gcgtcccact tgagccctcg gaagaagaga ccccggcaga cgggtgcctt
      421 gatggcctcc tctcctcaag acatcaaatt tcaagatttg gtcgtcttca ttttggagaa
      481 gaaaatggga accacccgca gagcgttcct catggagctg gcccgcagga aagggttcag
      541 ggttgaaaat gagctcagtg attctgtcac ccacattgta gcagagaaca actcgggttc
      601 ggatgttctg gagtggcttc aagcacagaa agtacaagtc agctcacaac cagagctcct
      661 cgatgtctcc tggctgatcg aatgcatagg agcagggaaa ccggtggaaa tgacaggaaa
      721 acaccagctt gttgtgagaa gagactattc agatagcacc aacccaggcc ccccgaagac
      781 tccaccaatt gctgtacaaa agatctccca gtatgcgtgt cagagaagaa ccactttaaa
      841 caactgtaac cagatattca cggatgcctt tgatatactg gctgaaaact gtgagtttag
      901 agaaaatgaa gactcctgtg tgacatttat gagagcagct tctgtattga aatctctgcc
      961 attcacaatc atcagtatga aggacacaga aggaattccc tgcctggggt ccaaggtgaa
     1021 gggtatcata gaggagatta ttgaagatgg agaaagttct gaagttaaag ctgtgttaaa
     1081 tgatgaacga tatcaatcct tcaaactctt tacttctgta tttggagtgg ggctgaagac
     1141 ttctgagaag tggttcagga tgggtttcag aactctgagt aaagtaaggt cggacaaaag
     1201 cctgaaattt acacgaatgc agaaagcagg atttctgtat tatgaagacc ttgtcagctg
     1261 tgtgaccagg gcagaagcag aggccgtcag tgtgctggtt aaagaggctg tctgggcatt
     1321 tcttccggat gctttcgtca ccatgacagg agggttccgg aggggtaaga agatggggca
     1381 tgatgtagat tttttaatta ccagcccagg atcaacagag gatgaagagc aacttttaca
     1441 gaaagtgatg aacttatggg aaaagaaggg attactttta tattatgacc ttgtggagtc
     1501 aacatttgaa aagctcaggt tgcctagcag gaaggttgat gctttggatc attttcaaaa
     1561 gtgctttctg attttcaaat tgcctcgtca aagagtggac agtgaccagt ccagctggca
     1621 ggaaggaaag acctggaagg ccatccgtgt ggatttagtt ctgtgcccct acgagcgtcg
     1681 tgcctttgcc ctgttgggat ggactggctc cattcagttt aagagagacc tcgcgcgcta
     1741 tgccacacat gagcggaaga tgattctgga taaccatgct ttatatgaca agaccaagag
     1801 gatattcctc aaagcagaaa gtgaagaaga aatttttgcg catctgggat tggattatat
     1861 tgaaccgtgg gaaagaaatg cctagtgaaa gggttcgatc cctaccggtt agtaatgagt
     1921 ttaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     1981 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     2041 ggggttcggt cccagggctg gagcagttac agagatgtta cgaaccctga acaggttgac
     2101 tattaattgt gtctgcttga tgtggacacc aggtggcgct ggacatcaga tttggagagg
     2161 cagttgtcta gggaaccggg ctctgtgcca gcgcaggagg caggctggct ctcctattcc
     2221 agggatgctc atccaggaag gaaaggttgc atgctggaca cactaacctt acagcggtgg
     2281 gctctatggg gtacctacgt atagatcttg agacaaatgg cagtattcat ccacaatttt
     2341 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2401 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     2461 tattacaggg acagcagaga tccactttgg cgccggctcg agtggctccg gtgcccgtca
     2521 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     2581 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     2641 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     2701 tctttttcgc aacgggtttg ccgccagaac acaggtgtcg tgacgcggga tccgccacca
     2761 tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg
     2821 atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta
     2881 aactgacgct gaagttcatc tgtactactg gtaaactgcc ggttccttgg ccgactctgg
     2941 taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc
     3001 atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta
     3061 aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa
     3121 accgcattga gctgaaaggc attgacttta aagaggacgg caatatcctg ggccataagc
     3181 tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca
     3241 ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc
     3301 actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc
     3361 tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc
     3421 tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgactcg
     3481 agcgacgcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3541 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3601 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3661 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatgagt
     3721 atcgatacta gttaaaagtt ttgttacttt atagaagaaa ttttgagttt ttgttttttt
     3781 ttaataaata aataaacata aataaattgt ttgttgaatt tattattagt atgtaagtgt
     3841 aaatataata aaacttaata tctattcaaa ttaataaata aacctcgata tacagaccga
     3901 taaaacacat gcgtcaattt tacgcatgat tatctttaac gtacgtcaca atatgattat
     3961 ctttctaggg ttaatctagt atacgcgtat gcggccgctt aattaatcca gcttttgttc
     4021 cctttagtga gggttaattc cgagcggccg cgtgttacaa ccaattaacc aattctgatt
     4081 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     4141 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     4201 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     4261 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     4321 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     4381 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc attattcatt cgtgattgcg
     4441 cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     4501 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     4561 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     4621 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     4681 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     4741 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     4801 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     4861 tggagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     4921 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     4981 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     5041 agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5101 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5161 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5221 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5281 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5341 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5401 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5461 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5521 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5581 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5641 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5701 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5761 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtgcgg ccgccaatta
     5821 atacgactca ctatagggcg aattggggcg cgccattcta gattaaccct agaaagatag
     5881 tctgcgtaaa attgacgcat gcattcttga aatattgctc tctctttcta aatagcgcga
     5941 atccgtcgct gtgcatttag gacatctcag tcgccgcttg gagctcccgt gaggcgtgct
     6001 tgtcaatgcg gtaagtgtca ctgattttga actataacga ccgcgtgagt caaaatgacg
     6061 catgattatc ttttacgtga cttttaagat ttaactcata cgataattat attgttattt
     6121 catgttctac ttacgtgata acttattata tatatatttt cttgttatag atatcaacta
     6181 gaatgctagc atgggcccag atctccatgg gccctgaatt cgcatctaga ctgaatcgat
     6241 aggtaccgag ctcttacgcg agatcaaagg gggtaagatc aaagggggta agatcaaagg
     6301 gggtaagatc aaagggggta agatcaaagg gggtaagatc aaagggggta agatcaaagg
     6361 ggcgcgcccg cgtgctagcc cgggctcgag atctagactc tagagggtat ataatggaag
     6421 ctcgaattcc agcttggcat tccggtactg ttggtaaaaa gcttggcatt ccggtactgt
     6481 tggtaaagcc accatggaca agaagtactc cattgggctc gatatcggca caaacagcgt
     6541 cggctgggcc gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg
     6601 caataccgat cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg
     6661 ggagacggcc gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa
     6721 gaatcggatc tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc
     6781 tttcttccat aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca
     6841 cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata
     6901 tcatctgagg aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct
     6961 cgcgctggcg catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc
     7021 agacaacagc gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt
     7081 cgaagagaac ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct
     7141 gtccaaatcc cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg
     7201 cctgtttggt aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt
     7261 cgacctggcc gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga
     7321 caatctgctg gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct
     7381 gtcagacgcc attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc
     7441 gctgagcgct agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa
     7501 ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa
     7561 aaatggctac gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat
     7621 taagcccatc ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga
     7681 agatctgttg cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct
     7741 gggcgaactg cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa
     7801 cagggaaaag attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc
     7861 ccggggaaat tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg
     7921 gaacttcgag gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac
     7981 taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga
     8041 gtacttcaca gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa
     8101 gccagcattc ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa
     8161 ccggaaagtt accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga
     8221 ctctgttgaa atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga
     8281 tctcctgaaa atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct
     8341 tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt
     8401 gaaaacttac gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata
     8461 tacaggatgg gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg
     8521 aaagacaatc ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt
     8581 gatccatgat gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca
     8641 gggggacagt cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg
     8701 aatactgcag accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc
     8761 cgagaatatc gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa
     8821 cagtagggaa aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct
     8881 taaggaacac ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct
     8941 gcagaacggc agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta
     9001 cgacgtggat catatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt
     9061 gttgacaaga tccgataaaa atagagggaa gagtgataac gtcccctcag aagaagttgt
     9121 caagaaaatg aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa
     9181 gttcgataat ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt
     9241 catcaaaagg cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga
     9301 ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat
     9361 tactctgaag tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag
     9421 agagatcaac aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc
     9481 acttatcaaa aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta
     9541 cgatgttagg aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta
     9601 cttcttttac agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga
     9661 gattcggaag cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa
     9721 gggtagggat ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa
     9781 aaagaccgaa gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag
     9841 cgacaagctg atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc
     9901 tcctacagtc gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa
     9961 actcaaaagc gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa
    10021 aaaccccatc gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat
    10081 taagcttccc aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag
    10141 tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt
    10201 gtatctggcc agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca
    10261 gctgttcgtg gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt
    10321 ctccaaaaga gtgatcct
//
