LOCUS       sequence-262603-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hypoxia-inducible Cas9 and TdT in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  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 13012)
  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: pTBL716 4xHRE-YB-TATA-Cas9-ODD-T2A-TdT
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    105..606
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     regulatory      1259..1268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1283..5386
                     /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             1283..5386
                     /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             1283..5386
                     /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             5399..5419
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5816..5836
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5846..5899
                     /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"
     misc_feature    7431..8019
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7484..7504)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7902..7913)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     enhancer        8075..8453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8454..8657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8608..8628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8702..8721
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8702..8720
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      8750..8759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8756..9154
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    9314..9435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9351..9370)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9405..9424
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    9748..10277
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(10627..10643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10627..10643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10640..10662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10651..10667
                     /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(10675..10681,10682..10699,10700..10705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10720..10741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10858..10875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11029..11617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11109..11128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11788..12579,12580..12648))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     12411..12430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12649..12753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12828..12847)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttggtg agtgaattcc tgccgcctcc ttcaggttcc gtcttcctcc actccctctt
       61 ccccttgctc tctgctgtgt tgctgcccaa ggatgctctt tccggagcac ttccttctcg
      121 gcgctgcacc acgtgatgtc ctctgagcgg atcctccccg tgtctgggtc ctctccgggc
      181 atctctcctc cctcacccaa ccccatgccg tcttcactcg ctgggttccc ttttccttct
      241 ccttctgggg cctgtgccat ctctcgtttc ttaggatggc cttctccgac ggatgtctcc
      301 cttgcgtccc gcctcccctt cttgtaggcc tgcatcatca ccgtttttct ggacaacccc
      361 aaagtacccc gtctccctgg ctttagccac ctctccatcc tcttgctttc tttgcctgga
      421 caccccgttc tcctgtggat tcgggtcacc tctcactcct ttcatttggg cagctcccct
      481 acccccctta cctctctagt ctgtgctagc tcttccagcc ccctgtcatg gcatcttcca
      541 ggggtccgag agctcagcta gtcttcttcc tccaacccgg gcccctatgt ccacttcagg
      601 acagcatgtt tgctgcctcc agggatcctg tgtccccgag ctgggaccac cttatattcc
      661 cagggccggt taatgtggct ctggttctgg gtacttttat ctgtcccctc caccccacag
      721 tggggccact agggacagga ttggtgacag aaaagcccca tccttaggca gctagtcagg
      781 tagtaaaagg tctcaactag gcagtggcag agcaggattc aaattcaggg ctgttgtgat
      841 gcctccgcag actctgagcg ccacctggtg gtaatttgtc tgtgcctctt ctgacgtgga
      901 agaacagcaa ctaacacact aacacggcat ttactatggg ccagccattg ttctaaagcc
      961 ctcgttcaca gagaaagttc ggttacactg cggattgacg gatcgggaga tctttgtatt
     1021 taattaagac cttgagtacg tgcgtctctg cacgtatggc gattaagacc ttgagtacgt
     1081 gcgtctctgc acgtatggcg attaagacct tgagtacgtg cgtctctgca cgtatggcga
     1141 ttaagacctt gagtacgtgc gtctctgcac gtatggcgat taatccatat gctctagagg
     1201 gtatataatg ggggccacta gtctactacc agaaagcttg gtaccgagct cggatccagc
     1261 caccatgggc cctagtggcc caatggacaa gaagtactcc attgggctcg atatcggcac
     1321 aaacagcgtc ggctgggccg tcattacgga cgagtacaag gtgccgagca aaaaattcaa
     1381 agttctgggc aataccgatc gccacagcat aaagaagaac ctcattggcg ccctcctgtt
     1441 cgactccggg gagacggccg aagccacgcg gctcaaaaga acagcacggc gcagatatac
     1501 ccgcagaaag aatcggatct gctacctgca ggagatcttt agtaatgaga tggctaaggt
     1561 ggatgactct ttcttccata ggctggagga gtcctttttg gtggaggagg ataaaaagca
     1621 cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc
     1681 aaccatatat catctgagga agaagcttgt agacagtact gataaggctg acttgcggtt
     1741 gatctatctc gcgctggcgc atatgatcaa atttcgggga cacttcctca tcgaggggga
     1801 cctgaaccca gacaacagcg atgtcgacaa actctttatc caactggttc agacttacaa
     1861 tcagcttttc gaagagaacc cgatcaacgc atccggagtt gacgccaaag caatcctgag
     1921 cgctaggctg tccaaatccc ggcggctcga aaacctcatc gcacagctcc ctggggagaa
     1981 gaagaacggc ctgtttggta atcttatcgc cctgtcactc gggctgaccc ccaactttaa
     2041 atctaacttc gacctggccg aagatgccaa gcttcaactg agcaaagaca cctacgatga
     2101 tgatctcgac aatctgctgg cccagatcgg cgaccagtac gcagaccttt ttttggcggc
     2161 aaagaacctg tcagacgcca ttctgctgag tgatattctg cgagtgaaca cggagatcac
     2221 caaagctccg ctgagcgcta gtatgatcaa gcgctatgat gagcaccacc aagacttgac
     2281 tttgctgaag gcccttgtca gacagcaact gcctgagaag tacaaggaaa ttttcttcga
     2341 tcagtctaaa aatggctacg ccggatacat tgacggcgga gcaagccagg aggaatttta
     2401 caaatttatt aagcccatct tggaaaaaat ggacggcacc gaggagctgc tggtaaagct
     2461 taacagagaa gatctgttgc gcaaacagcg cactttcgac aatggaagca tcccccacca
     2521 gattcacctg ggcgaactgc acgctatcct caggcggcaa gaggatttct accccttttt
     2581 gaaagataac agggaaaaga ttgagaaaat cctcacattt cggataccct actatgtagg
     2641 ccccctcgcc cggggaaatt ccagattcgc gtggatgact cgcaaatcag aagagaccat
     2701 cactccctgg aacttcgagg aagtcgtgga taagggggcc tctgcccagt ccttcatcga
     2761 aaggatgact aactttgata aaaatctgcc taacgaaaag gtgcttccta aacactctct
     2821 gctgtacgag tacttcacag tttataacga gctcaccaag gtcaaatacg tcacagaagg
     2881 gatgagaaag ccagcattcc tgtctggaga gcagaagaaa gctatcgtgg acctcctctt
     2941 caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa gactatttca aaaagattga
     3001 atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac
     3061 gtatcacgat ctcctgaaaa tcattaaaga caaggacttc ctggacaatg aggagaacga
     3121 ggacattctt gaggacattg tcctcaccct tacgttgttt gaagataggg agatgattga
     3181 agaacgcttg aaaacttacg ctcatctctt cgacgacaaa gtcatgaaac agctcaagag
     3241 gcgccgatat acaggatggg ggcggctgtc aagaaaactg atcaatggga tccgagacaa
     3301 gcagagtgga aagacaatcc tggattttct taagtccgat ggatttgcca accggaactt
     3361 catgcagttg atccatgatg actctctcac ctttaaggag gacatccaga aagcacaagt
     3421 ttctggccag ggggacagtc ttcacgagca catcgctaat cttgcaggta gcccagctat
     3481 caaaaaggga atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag
     3541 gcataagccc gagaatatcg ttatcgagat ggcccgagag aaccaaacta cccagaaggg
     3601 acagaagaac agtagggaaa ggatgaagag gattgaagag ggtataaaag aactggggtc
     3661 ccaaatcctt aaggaacacc cagttgaaaa cacccagctt cagaatgaga agctctacct
     3721 gtactacctg cagaacggca gggacatgta cgtggatcag gaactggaca tcaatcggct
     3781 ctccgactac gacgtggatc atatcgtgcc ccagtctttt ctcaaagatg attctattga
     3841 taataaagtg ttgacaagat ccgataaaaa tagagggaag agtgataacg tcccctcaga
     3901 agaagttgtc aagaaaatga aaaattattg gcggcagctg ctgaacgcca aactgatcac
     3961 acaacggaag ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg agttggataa
     4021 agccggcttc atcaaaaggc agcttgttga gacacgccag atcaccaagc acgtggccca
     4081 aattctcgat tcacgcatga acaccaagta cgatgaaaat gacaaactga ttcgagaggt
     4141 gaaagttatt actctgaagt ctaagctggt ctcagatttc agaaaggact ttcagtttta
     4201 taaggtgaga gagatcaaca attaccacca tgcgcatgat gcctacctga atgcagtggt
     4261 aggcactgca cttatcaaaa aatatcccaa gcttgaatct gaatttgttt acggagacta
     4321 taaagtgtac gatgttagga aaatgatcgc aaagtctgag caggaaatag gcaaggccac
     4381 cgctaagtac ttcttttaca gcaatattat gaattttttc aagaccgaga ttacactggc
     4441 caatggagag attcggaagc gaccacttat cgaaacaaac ggagaaacag gagaaatcgt
     4501 gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa
     4561 catcgttaaa aagaccgaag tacagaccgg aggcttctcc aaggaaagta tcctcccgaa
     4621 aaggaacagc gacaagctga tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg
     4681 attcgattct cctacagtcg cttacagtgt actggttgtg gccaaagtgg agaaagggaa
     4741 gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg agcgatcaag
     4801 cttcgaaaaa aaccccatcg actttctcga ggcgaaagga tataaagagg tcaaaaaaga
     4861 cctcatcatt aagcttccca agtactctct ctttgagctt gaaaacggcc ggaaacgaat
     4921 gctcgctagt gcgggcgagc tgcagaaagg taacgagctg gcactgccct ctaaatacgt
     4981 taatttcttg tatctggcca gccactatga aaagctcaaa gggtctcccg aagataatga
     5041 gcagaagcag ctgttcgtgg aacaacacaa acactacctt gatgagatca tcgagcaaat
     5101 aagcgaattc tccaaaagag tgatcctcgc cgacgctaac ctcgataagg tgctttctgc
     5161 ttacaataag cacagggata agcccatcag ggagcaggca gaaaacatta tccacttgtt
     5221 tactctgacc aacttgggcg cgcctgcagc cttcaagtac ttcgacacca ccatagacag
     5281 aaagcggtac acctctacaa aggaggtcct ggacgccaca ctgattcatc agtcaattac
     5341 ggggctctat gaaacaagaa tcgacctctc tcagctcggt ggagacagca gggctgaccc
     5401 caagaagaag aggaaggtgg gcgggggagg atccgaattc aagttggaat tggtagaaaa
     5461 actttttgct gaagacacag aagcaaagaa cccattttct actcaggaca cagatttaga
     5521 cttggagatg ttagctccct atatcccaat ggatgatgac ttccagttac gttccttcga
     5581 tcagttgtca ccattagaaa gcagttccgc aagccctgaa agcgcaagtc ctcaaagcac
     5641 agttacagta ttccagcaga ctcaaataca agaacctact gctaatgcca ccactaccac
     5701 tgccaccact gatgaattaa aaacagtgac aaaagaccgt atggaagaca ttaaaatatt
     5761 gattgcatct ccatctccta cccacataca taaagaaact acttctggtg gttctcccaa
     5821 gaagaagagg aaagtcggca gcggcgaggg cagaggcagc ctgctgacct gcggcgacgt
     5881 ggaggagaac cccggcccca tggatccacc acgagcgtcc cacttgagcc ctcggaagaa
     5941 gagaccccgg cagacgggtg ccttgatggc ctcctctcct caagacatca aatttcaaga
     6001 tttggtcgtc ttcattttgg agaagaaaat gggaaccacc cgcagagcgt tcctcatgga
     6061 gctggcccgc aggaaagggt tcagggttga aaatgagctc agtgattctg tcacccacat
     6121 tgtagcagag aacaactcgg gttcggatgt tctggagtgg cttcaagcac agaaagtaca
     6181 agtcagctca caaccagagc tcctcgatgt ctcctggctg atcgaatgca taggagcagg
     6241 gaaaccggtg gaaatgacag gaaaacacca gcttgttgtg agaagagact attcagatag
     6301 caccaaccca ggccccccga agactccacc aattgctgta caaaagatct cccagtatgc
     6361 gtgtcagaga agaaccactt taaacaactg taaccagata ttcacggatg cctttgatat
     6421 actggctgaa aactgtgagt ttagagaaaa tgaagactcc tgtgtgacat ttatgagagc
     6481 agcttctgta ttgaaatctc tgccattcac aatcatcagt atgaaggaca cagaaggaat
     6541 tccctgcctg gggtccaagg tgaagggtat catagaggag attattgaag atggagaaag
     6601 ttctgaagtt aaagctgtgt taaatgatga acgatatcaa tccttcaaac tctttacttc
     6661 tgtatttgga gtggggctga agacttctga gaagtggttc aggatgggtt tcagaactct
     6721 gagtaaagta aggtcggaca aaagcctgaa atttacacga atgcagaaag caggatttct
     6781 gtattatgaa gaccttgtaa gctgtgtgac cagggcagaa gcagaggccg tcagtgtgct
     6841 ggttaaagag gctgtctggg catttcttcc ggatgctttc gtcaccatga caggagggtt
     6901 ccggaggggt aagaagatgg ggcatgatgt agatttttta attaccagcc caggatcaac
     6961 agaggatgaa gagcaacttt tacagaaagt gatgaactta tgggaaaaga agggattact
     7021 tttatattat gaccttgtgg agtcaacatt tgaaaagctc aggttgccta gcaggaaggt
     7081 tgatgctttg gatcattttc aaaagtgctt tctgattttc aaattgcctc gtcaaagagt
     7141 ggacagtgac cagtccagct ggcaggaagg aaagacctgg aaggccatcc gtgtggattt
     7201 agttctgtgc ccctacgagc gtcgtgcctt tgccctgttg ggatggactg gctcccggca
     7261 gtttgagaga gacctccggc gctatgccac acatgagcgg aagatgattc tggataacca
     7321 tgctttatat gacaagacca agaggatatt cctcaaagca gaaagtgaag aagaaatttt
     7381 tgcgcatctg ggattggatt atattgaacc gtgggaaaga aatgcctagc aatcaacctc
     7441 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     7501 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     7561 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     7621 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     7681 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     7741 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     7801 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     7861 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     7921 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     7981 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattctg cagatatccg
     8041 gttagtaatg agtttggaat taattctgtg gaatacattg attattgact agttattaat
     8101 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     8161 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     8221 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     8281 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     8341 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     8401 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     8461 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     8521 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     8581 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     8641 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     8701 ttaatacgac tcactatagg gagacccaag ctggctagcg ctaccggtcg ccaccatggc
     8761 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
     8821 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
     8881 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
     8941 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
     9001 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
     9061 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
     9121 attgctgccc tctggttatg tgtgggaggg ctaattcgaa atgaccgacc aagcgacgcc
     9181 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     9241 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     9301 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     9361 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     9421 catcaatgta tcttatcatg tctgtataag ctagtcaggt agtaaaaggt ctcaactagg
     9481 cagtggcaga gcaggattca aattcagggc tgttgtgatg cctccgcaga ctctgagcgc
     9541 cacctggtgg taatttgtct gtgcctcttc tgacgtggaa gaacagcaac taacacacta
     9601 acacggcatt tactatgggc cagccattgt tctaattggg tctaaccccc acctcctgtt
     9661 aggcagattc cttatctggt gacacacccc catttcctgg agccatctct ctccttgcca
     9721 gaacctctaa ggtttgctta cgatggagcc agagaggatc ctgggaggga gagcttggca
     9781 gggggtggga gggaaggggg ggatgcgtga cctgcccggt tctcagtggc caccctgcgc
     9841 taccctctcc cagaacctga gctgctctga cgcggctgtc tggtgcgttt cactgatcct
     9901 ggtgctgcag cttccttaca cttcccaaga ggagaagcag tttggaaaaa caaaatcaga
     9961 ataagttggt cctgagttct aactttggct cttcaccttt ctagtcccca atttatattg
    10021 ttcctccgtg cgtcagtttt acctgtgaga taaggccagt agccagcccc gtcctggcag
    10081 ggctgtggtg aggagggggg tgtccgtgtg gaaaactccc tttgtgagaa tggtgcgtcc
    10141 taggtgttca ccaggtcgtg gccgcctcta ctccctttct ctttctccat ccttctttcc
    10201 ttaaagagtc cccagtgcta tctgggacat attcctccgc ccagagcagg gtcccgcttc
    10261 cctaaggccc tgctctgggc ttctgggttt gagtccttgg caagcccagg agaggcgctc
    10321 aggcttccct gtcccccttc ctcgtccacc atctcatgcc cctggctctc ctgccccttc
    10381 cctacagggg ttcctggctc tgctcttcag actgagcccc gttcccctgc atccccgttc
    10441 ccctgcatcc cccttcccct gcatccccca gaggccccag gccacctact tggcctggac
    10501 cccacgagag gccaccccag ccctgtctac caggctgcct tttgggtgga ttctcctcca
    10561 actgtggggt gactgcttgg caaacgaatt cccgtcgacc tctagctaga gcttggcgta
    10621 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
    10681 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
    10741 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
    10801 atgaatcggc caacgcgcgg ggagaggcgg tttgtgtatt gggcgctctt ccgcttcctc
    10861 gctcactgac tcgctgcgct cggtcgttcg gctgtggcga gcggtatcag ctcactcaaa
    10921 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
    10981 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
    11041 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
    11101 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
    11161 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
    11221 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
    11281 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
    11341 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
    11401 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
    11461 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    11521 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    11581 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    11641 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    11701 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    11761 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    11821 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    11881 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
    11941 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    12001 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    12061 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    12121 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    12181 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    12241 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    12301 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    12361 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    12421 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    12481 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    12541 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    12601 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    12661 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    12721 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    12781 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
    12841 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
    12901 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
    12961 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata ta
//
