LOCUS       sequence-449157-       10834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBAC 4xHRE YB_TATA hCas9-T2A-G/C-biased1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10834)
  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 10834)
  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: pCKC776
FEATURES             Location/Qualifiers
     source          1..10834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1539..1558)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1583..1631
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    1936..2053
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2122..2333
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2353..2362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2359..3066
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2504..2522)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2772..2791
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3099..3116)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3105..3312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3519..3581
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(3632..3650)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(3690..4505)
                     /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(3690..4505)
                     /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     4413..4432
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4497..4516
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4730..5318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5219..5238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5430..5449
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5430..5448
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5478..5512
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     regulatory      6148..6157
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6172..10275
                     /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             6172..10275
                     /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             6172..10275
                     /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             10288..10308
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10705..10725
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10735..10788
                     /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"
ORIGIN
        1 cccggcagac gggtgccttg atggcctcct ctcctcaaga catcaaattt caagatttgg
       61 tcgtcttcat tttggagaag aaaatgggaa ccacccgcag agcgttcctc atggagctgg
      121 cccgcaggaa agggttcagg gttgaaaatg agctcagtga ttctgtcacc cacattgtag
      181 cagagaacaa ctcgggttcg gatgttctgg agtggcttca agcacagaaa gtacaagtca
      241 gctcacaacc agagctcctc gatgtctcct ggctgatcga atgcatagga gcagggaaac
      301 cggtggaaat gacaggaaaa caccagcttg ttgtgagaag agactattca gatagcacca
      361 acccaggccc cccgaagact ccaccaattg ctgtacaaaa gatctcccag tatgcgtgtc
      421 agagaagaac cactttaaac aactgtaacc agatattcac ggatgccttt gatatactgg
      481 ctgaaaactg tgagtttaga gaaaatgaag actcctgtgt gacatttatg agagcagctt
      541 ctgtattgaa atctctgcca ttcacaatca tcagtatgaa ggacacagaa ggaattccct
      601 gcctggggtc caaggtgaag ggtatcatag aggagattat tgaagatgga gaaagttctg
      661 aagttaaagc tgtgttaaat gatgaacgat atcaatcctt caaactcttt acttctgtat
      721 ttggagtggg gctgaagact tctgagaagt ggttcaggat gggtttcaga actctgagta
      781 aagtaaggtc ggacaaaagc ctgaaattta cacgaatgca gaaagcagga tttctgtatt
      841 atgaagacct tgtcagctgt gtgaccaggg cagaagcaga ggccgtcagt gtgctggtta
      901 aagaggctgt ctgggcattt cttccggatg ctttcgtcac catgacagga gggttccgga
      961 ggggtaagaa gatggggcat gatgtagatt ttttaattac cagcccagga tcaacagagg
     1021 atgaagagca acttttacag aaagtgatga acttatggga aaagaaggga ttacttttat
     1081 attatgacct tgtggagtca acatttgaaa agctcaggtt gcctagcagg aaggttgatg
     1141 ctatggatca ttttcaaaag tgctttctga ttttcaaatt gcctcgtcaa agagtggaca
     1201 gtgaccagtc cagctggcag gaaggaaaga cctggaaggc catccgtgtg gatttagttc
     1261 tgtgccccta cgagcgtcgt gcctttgccc tgttgggatg gactggatcc gatcagtttg
     1321 atagagacct ccggcgctat gccacacatg agcggaagat gattctggat aaccatgctt
     1381 tatatgacaa gaccaagagg atattcctca aagcagaaag tgaagaagaa atttttgcgc
     1441 atctgggatt ggattatatt gaaccgtggg aaagaaatgc ctagtgaaag ggttcgatcc
     1501 ctaccggtta gtaatgagtt taaacggggg aggctaactg aaacacggaa ggagacaata
     1561 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     1621 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg agcagttaca gagatgttac
     1681 gaaccctgaa caggttgact attaattgtg tctgcttgat gtggacacca ggtggcgctg
     1741 gacatcagat ttggagaggc agttgtctag ggaaccgggc tctgtgccag cgcaggaggc
     1801 aggctggctc tcctattcca gggatgctca tccaggaagg aaaggttgca tgctggacac
     1861 actaacctta cagcggtggg ctctatgggg tacctacgta tagatcttga gacaaatggc
     1921 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     1981 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     2041 aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga
     2101 gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2161 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2221 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2281 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtgtcgt
     2341 gacgcgggat ccgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     2401 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     2461 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     2521 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     2581 aaggcctacg tgaagcaccc cgccgacatt cccgactact tgaagctgtc cttccccgag
     2641 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     2701 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     2761 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     2821 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     2881 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     2941 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     3001 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     3061 tacaagtcta cagatgtcga atcctaactc gagcgacgcc tcgactgtgc cttctagttg
     3121 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3181 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3241 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3301 gcatgctggg gatgcggtgg gctctatgag tatcgatact agttaaaagt tttgttactt
     3361 tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat aaataaattg
     3421 tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa
     3481 attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacgcatga
     3541 ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaatctag tatacgcgta
     3601 tgcggccgct taattaatcc agcttttgtt ccctttagtg agggttaatt ccgagcggcc
     3661 gcgtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
     3721 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     3781 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     3841 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     3901 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     3961 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     4021 tcaaccaaac cattattcat tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg
     4081 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     4141 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     4201 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     4261 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     4321 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     4381 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     4441 tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg
     4501 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     4561 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga
     4621 ataaatcgaa cttttgctga gttgaaggat cagtcatgac caaaatccct taacgtgagt
     4681 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4801 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4861 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4921 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4981 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5041 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5101 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5161 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5221 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5281 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5341 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5401 attctgtgga taaccgtgcg gccgccaatt aatacgactc actatagggc gaattggggc
     5461 gcgccattct agattaaccc tagaaagata gtctgcgtaa aattgacgca tgcattcttg
     5521 aaatattgct ctctctttct aaatagcgcg aatccgtcgc tgtgcattta ggacatctca
     5581 gtcgccgctt ggagctcccg tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg
     5641 aactataacg accgcgtgag tcaaaatgac gcatgattat cttttacgtg acttttaaga
     5701 tttaactcat acgataatta tattgttatt tcatgttcta cttacgtgat aacttattat
     5761 atatatattt tcttgttata gatatcaact agaatgctag catgggccca gatctccatg
     5821 ggccctgaat tcgcatctag actgaagccc tcgttcacag agaaagttcg gttacactgc
     5881 ggattgacgg atcgggagat ctttgtattt aattaagacc ttgagtacgt gcgtctctgc
     5941 acgtatggcg attaagacct tgagtacgtg cgtctctgca cgtatggcga ttaagacctt
     6001 gagtacgtgc gtctctgcac gtatggcgat taagaccttg agtacgtgcg tctctgcacg
     6061 tatggcgatt aatccatatg ctctagaggg tatataatgg gggccactag tctactacca
     6121 gaaagcttgg taccgagctc ggatccagcc accatgggcc ctagtggccc aatggacaag
     6181 aagtactcca ttgggctcga tatcggcaca aacagcgtcg gctgggccgt cattacggac
     6241 gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg ccacagcata
     6301 aagaagaacc tcattggcgc cctcctgttc gactccgggg agacggccga agccacgcgg
     6361 ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg ctacctgcag
     6421 gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag gctggaggag
     6481 tcctttttgg tggaggagga taaaaagcac gagcgccacc caatctttgg caatatcgtg
     6541 gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa gaagcttgta
     6601 gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca tatgatcaaa
     6661 tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga tgtcgacaaa
     6721 ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc gatcaacgca
     6781 tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa
     6841 aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa tcttatcgcc
     6901 ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga agatgccaag
     6961 cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc ccagatcggc
     7021 gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat tctgctgagt
     7081 gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag tatgatcaag
     7141 cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag acagcaactg
     7201 cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc cggatacatt
     7261 gacggcggag caagccagga ggaattttac aaatttatta agcccatctt ggaaaaaatg
     7321 gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg caaacagcgc
     7381 actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca cgctatcctc
     7441 aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat tgagaaaatc
     7501 ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc cagattcgcg
     7561 tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga agtcgtggat
     7621 aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa aaatctgcct
     7681 aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt ttataacgag
     7741 ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct gtctggagag
     7801 cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac cgtgaaacag
     7861 ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat cagcggagtg
     7921 gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat cattaaagac
     7981 aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt cctcaccctt
     8041 acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc tcatctcttc
     8101 gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg gcggctgtca
     8161 agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct ggattttctt
     8221 aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga ctctctcacc
     8281 tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct tcacgagcac
     8341 atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac cgttaaggtc
     8401 gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt tatcgagatg
     8461 gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag gatgaagagg
     8521 attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc agttgaaaac
     8581 acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag ggacatgtac
     8641 gtggatcagg aactggacat caatcggctc tccgactacg acgtggatca tatcgtgccc
     8701 cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc cgataaaaat
     8761 agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa aaattattgg
     8821 cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct gactaaggct
     8881 gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca gcttgttgag
     8941 acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa caccaagtac
     9001 gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc taagctggtc
     9061 tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa ttaccaccat
     9121 gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa atatcccaag
     9181 cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa aatgatcgca
     9241 aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag caatattatg
     9301 aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg accacttatc
     9361 gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt cgcgacagtc
     9421 cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt acagaccgga
     9481 ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa
     9541 aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc ttacagtgta
     9601 ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg
     9661 ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag
     9721 gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa gtactctctc
     9781 tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt
     9841 aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag ccactatgaa
     9901 aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga acaacacaaa
     9961 cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt gatcctcgcc
    10021 gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa gcccatcagg
    10081 gagcaggcag aaaacattat ccacttgttt actctgacca acttgggcgc gcctgcagcc
    10141 ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa ggaggtcctg
    10201 gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat cgacctctct
    10261 cagctcggtg gagacagcag ggctgacccc aagaagaaga ggaaggtggg cgggggagga
    10321 tccgaattca agttggaatt ggtagaaaaa ctttttgctg aagacacaga agcaaagaac
    10381 ccattttcta ctcaggacac agatttagac ttggagatgt tagctcccta tatcccaatg
    10441 gatgatgact tccagttacg ttccttcgat cagttgtcac cattagaaag cagttccgca
    10501 agccctgaaa gcgcaagtcc tcaaagcaca gttacagtat tccagcagac tcaaatacaa
    10561 gaacctactg ctaatgccac cactaccact gccaccactg atgaattaaa aacagtgaca
    10621 aaagaccgta tggaagacat taaaatattg attgcatctc catctcctac ccacatacat
    10681 aaagaaacta cttctggtgg ttctcccaag aagaagagga aagtcggcag cggcgagggc
    10741 agaggcagcc tgctgacctg cggcgacgtg gaggagaacc ccggccccat ggatccacca
    10801 cgagcgtccc acttgagccc tcggaagaag agac
//
