LOCUS       sequence-248550-       11256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9-T2A-TdT in mammalian cells. (pTBL209).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11256)
  AUTHORS   Loveless TB, Grotts JH, Schechter MW, Forouzmand E, Carlson CK, 
            Agahi BS, Liang G, Ficht M, Liu B, Xie X, Liu CC
  TITLE     Lineage tracing and analog recording in mammalian cells by 
            single-site DNA writing.
  JOURNAL   Nat Chem Biol. 2021 Jun;17(6):739-747. doi: 
            10.1038/s41589-021-00769-8. Epub 2021 Mar 22.
  PUBMED    33753928
REFERENCE   2  (bases 1 to 11256)
  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"; doi: 
            "10.1038/s41589-021-00769-8"; journalName: "Nat Chem Biol"; date: 
            "2021-06"; volume: "17"; issue: "6"; pages: "739-747"
COMMENT     Sequence Label: pcDNA-Cas9-T2A-TdT
FEATURES             Location/Qualifiers
     source          1..11256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      911..920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             917..5020
                     /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             917..5020
                     /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             917..5020
                     /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             5033..5053
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5063..5116
                     /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(6691..6708)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6697..6921
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6967..7395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7054..7073)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7264..7285
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7404..7424)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7409..7738
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7589..7724
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7651..7670
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    8929..9050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8966..8985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9020..9039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9099..9115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9099..9115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9112..9134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9123..9139
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9147..9153,9154..9171,9172..9177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9192..9213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9330..9347)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9501..10089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9581..9600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10260..11051,11052..11120))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGALVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10883..10902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11121..11225)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cctgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gctaccggtc gccaccatgg acaagaagta ctccattggg ctcgatatcg gcacaaacag
      961 cgtcggctgg gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct
     1021 gggcaatacc gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc
     1081 cggggagacg gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag
     1141 aaagaatcgg atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga
     1201 ctctttcttc cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg
     1261 ccacccaatc tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat
     1321 atatcatctg aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta
     1381 tctcgcgctg gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa
     1441 cccagacaac agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct
     1501 tttcgaagag aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag
     1561 gctgtccaaa tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa
     1621 cggcctgttt ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa
     1681 cttcgacctg gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct
     1741 cgacaatctg ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa
     1801 cctgtcagac gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc
     1861 tccgctgagc gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct
     1921 gaaggccctt gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc
     1981 taaaaatggc tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt
     2041 tattaagccc atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag
     2101 agaagatctg ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca
     2161 cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga
     2221 taacagggaa aagattgaga aaatcctcac atttcggata ccctactatg taggccccct
     2281 cgcccgggga aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc
     2341 ctggaacttc gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat
     2401 gactaacttt gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta
     2461 cgagtacttc acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag
     2521 aaagccagca ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac
     2581 gaaccggaaa gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt
     2641 cgactctgtt gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca
     2701 cgatctcctg aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat
     2761 tcttgaggac attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg
     2821 cttgaaaact tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg
     2881 atatacagga tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag
     2941 tggaaagaca atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca
     3001 gttgatccat gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg
     3061 ccagggggac agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa
     3121 gggaatactg cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa
     3181 gcccgagaat atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa
     3241 gaacagtagg gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat
     3301 ccttaaggaa cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta
     3361 cctgcagaac ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga
     3421 ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa
     3481 agtgttgaca agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt
     3541 tgtcaagaaa atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg
     3601 gaagttcgat aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg
     3661 cttcatcaaa aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct
     3721 cgattcacgc atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt
     3781 tattactctg aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt
     3841 gagagagatc aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac
     3901 tgcacttatc aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt
     3961 gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa
     4021 gtacttcttt tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg
     4081 agagattcgg aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga
     4141 caagggtagg gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt
     4201 taaaaagacc gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa
     4261 cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga
     4321 ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa
     4381 aaaactcaaa agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga
     4441 aaaaaacccc atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat
     4501 cattaagctt cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc
     4561 tagtgcgggc gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt
     4621 cttgtatctg gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa
     4681 gcagctgttc gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga
     4741 attctccaaa agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa
     4801 taagcacagg gataagccca tcagggagca ggcagaaaac attatccact tgtttactct
     4861 gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg
     4921 gtacacctct acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct
     4981 ctatgaaaca agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa
     5041 gaagaggaag gtgggcagcg gcgagggcag aggcagcctg ctgacctgcg gcgacgtgga
     5101 ggagaacccc ggccccatgg atccaccacg agcgtcccac ttgagccctc ggaagaagag
     5161 accccggcag acgggtgcct tgatggcctc ctctcctcaa gacatcaaat ttcaagattt
     5221 ggtcgtcttc attttggaga agaaaatggg aaccacccgc agagcgttcc tcatggagct
     5281 ggcccgcagg aaagggttca gggttgaaaa tgagctcagt gattctgtca cccacattgt
     5341 agcagagaac aactcgggtt cggatgttct ggagtggctt caagcacaga aagtacaagt
     5401 cagctcacaa ccagagctcc tcgatgtctc ctggctgatc gaatgcatag gagcagggaa
     5461 accggtggaa atgacaggaa aacaccagct tgttgtgaga agagactatt cagatagcac
     5521 caacccaggc cccccgaaga ctccaccaat tgctgtacaa aagatctccc agtatgcgtg
     5581 tcagagaaga accactttaa acaactgtaa ccagatattc acggatgcct ttgatatact
     5641 ggctgaaaac tgtgagttta gagaaaatga agactcctgt gtgacattta tgagagcagc
     5701 ttctgtattg aaatctctgc cattcacaat catcagtatg aaggacacag aaggaattcc
     5761 ctgcctgggg tccaaggtga agggtatcat agaggagatt attgaagatg gagaaagttc
     5821 tgaagttaaa gctgtgttaa atgatgaacg atatcaatcc ttcaaactct ttacttctgt
     5881 atttggagtg gggctgaaga cttctgagaa gtggttcagg atgggtttca gaactctgag
     5941 taaagtaagg tcggacaaaa gcctgaaatt tacacgaatg cagaaagcag gatttctgta
     6001 ttatgaagac cttgtaagct gtgtgaccag ggcagaagca gaggccgtca gtgtgctggt
     6061 taaagaggct gtctgggcat ttcttccgga tgctttcgtc accatgacag gagggttccg
     6121 gaggggtaag aagatggggc atgatgtaga ttttttaatt accagcccag gatcaacaga
     6181 ggatgaagag caacttttac agaaagtgat gaacttatgg gaaaagaagg gattactttt
     6241 atattatgac cttgtggagt caacatttga aaagctcagg ttgcctagca ggaaggttga
     6301 tgctttggat cattttcaaa agtgctttct gattttcaaa ttgcctcgtc aaagagtgga
     6361 cagtgaccag tccagctggc aggaaggaaa gacctggaag gccatccgtg tggatttagt
     6421 tctgtgcccc tacgagcgtc gtgcctttgc cctgttggga tggactggct cccggcagtt
     6481 tgagagagac ctccggcgct atgccacaca tgagcggaag atgattctgg ataaccatgc
     6541 tttatatgac aagaccaaga ggatattcct caaagcagaa agtgaagaag aaatttttgc
     6601 gcatctggga ttggattata ttgaaccgtg ggaaagaaat gcctaggcgg ccgctcgagt
     6661 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     6721 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     6781 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     6841 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     6901 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     6961 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     7021 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     7081 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     7141 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     7201 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     7261 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     7321 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     7381 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     7441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     7501 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     7561 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     7621 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     7681 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     7741 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     7801 tcgcatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     7861 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     7921 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaatttag
     7981 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     8041 tgaaaccgaa ctgcccgctg ttctgcaacc ggtcgcggag gccatggatg cgatcgctgc
     8101 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     8161 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     8221 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     8281 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     8341 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     8401 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     8461 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     8521 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     8581 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     8641 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     8701 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataggcggg
     8761 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     8821 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     8881 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     8941 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     9001 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     9061 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     9121 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     9181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     9241 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     9301 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     9361 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     9421 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     9481 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     9541 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     9601 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     9661 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     9721 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     9781 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     9841 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     9901 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     9961 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
    10021 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
    10081 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
    10141 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
    10201 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
    10261 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    10321 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    10381 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    10441 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    10501 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    10561 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    10621 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    10681 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    10741 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    10801 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    10861 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    10921 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    10981 agttcgatgt aacccactag tgcacccaac tgatcttcag catcttttac tttcaccagc
    11041 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    11101 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    11161 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    11221 ccgcgcacat ttccccgaaa agtgccacct tacgtc
//
