LOCUS       sequence-367391-       10533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Cas9 nuclease and miRFP670 via T2A linker under 
            control of a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10533)
  AUTHORS   Abatti LE, Lado-Fernandez P, Huynh L, Collado M, Hoffman MM, 
            Mitchell JA
  TITLE     Epigenetic reprogramming of a distal developmental enhancer cluster 
            drives SOX2 overexpression in breast and lung adenocarcinoma.
  JOURNAL   Nucleic Acids Res. 2023 Sep 22:gkad734. doi: 10.1093/nar/gkad734.
  PUBMED    37738673
REFERENCE   2  (bases 1 to 10533)
  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: "Nucleic 
            Acids Res. 2023 Sep 22:gkad734. doi: 10.1093/nar/gkad734."
COMMENT     Sequence Label: CMV-hCas9-T2A-miRFP670
FEATURES             Location/Qualifiers
     source          1..10533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        173..552
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        553..756
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     707..727
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     753..777
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      852..861
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             858..4961
                     /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             858..4961
                     /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             858..4961
                     /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             4974..4994
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5001..5054
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5061..6005
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="VAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDHR
                     VIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTEY
                     CGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTGY
                     DRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVTY
                     QPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLVV
                     CHHYLPRFIRFELRAICKRLAERIATRITALES"
     primer_bind     complement(6042..6061)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6086..6134
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6336..6764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6423..6442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6633..6654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6773..6793)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6778..7107
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6958..7093
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7020..7039
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7174..7968
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7228..7247)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7838..7857
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8144..8265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8181..8200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8235..8254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8314..8330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8314..8330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8327..8349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8338..8354
                     /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(8362..8368,8369..8386,8387..8392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8407..8428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8545..8562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8716..9304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8796..8815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9475..10266,10267..10335))
                     /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     10098..10117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10336..10440)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(10515..10533,1))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
       61 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      121 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      181 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      241 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      301 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      361 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      421 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      481 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      541 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      601 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      661 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      721 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
      781 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
      841 cggactctag agccaccatg gacaagaagt actccattgg gctcgatatc ggcacaaaca
      901 gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
      961 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
     1021 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
     1081 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
     1141 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
     1201 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
     1261 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
     1321 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
     1381 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
     1441 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
     1501 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     1561 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     1621 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
     1681 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     1741 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     1801 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     1861 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     1921 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     1981 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
     2041 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     2101 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     2161 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     2221 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     2281 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     2341 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     2401 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     2461 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     2521 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     2581 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     2641 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     2701 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     2761 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
     2821 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     2881 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     2941 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     3001 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     3061 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     3121 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     3181 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     3241 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     3301 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     3361 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
     3421 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     3481 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     3541 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     3601 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     3661 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     3721 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     3781 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     3841 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     3901 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     3961 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     4021 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     4081 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     4141 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     4201 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     4261 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     4321 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     4381 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     4441 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     4501 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     4561 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     4621 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     4681 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     4741 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     4801 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     4861 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     4921 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
     4981 agaagaggaa ggtggctagc gagggcagag gaagtcttct aacatgcggt gacgtggagg
     5041 agaatcccgg ccctggtacc gtagcaggtc atgcctctgg cagccccgca ttcgggaccg
     5101 cctctcattc gaattgcgaa catgaagaga tccacctcgc cggctcgatc cagccgcatg
     5161 gcgcgcttct ggtcgtcagc gaacatgatc atcgcgtcat ccaggccagc gccaacgccg
     5221 cggaatttct gaatctcgga agcgtactcg gcgttccgct cgccgagatc gacggcgatc
     5281 tgttgatcaa gatcctgccg catctcgatc ccaccgccga aggcatgccg gtcgcggtgc
     5341 gctgccggat cggcaatccc tctacggagt actgcggtct gatgcatcgg cctccggaag
     5401 gcgggctgat catcgaactc gaacgtgccg gcccgtcgat cgatctgtca ggcacgctgg
     5461 cgccggcgct ggagcggatc cgcacggcgg gttcactgcg cgcgctgtgc gatgacaccg
     5521 tgctgctgtt tcagcagtgc accggctacg accgggtgat ggtgtatcgt ttcgatgagc
     5581 aaggccacgg cctggtattc tccgagtgcc atgtgcctgg gctcgaatcc tatttcggca
     5641 accgctatcc gtcgtcgact gtcccgcaga tggcgcggca gctgtacgtg cggcagcgcg
     5701 tccgcgtgct ggtcgacgtc acctatcagc cggtgccgct ggagccgcgg ctgtcgccgc
     5761 tgaccgggcg cgatctcgac atgtcgggct gcttcctgcg ctcgatgtcg ccgtgccatc
     5821 tgcagttcct gaaggacatg ggcgtgcgcg ccaccctggc ggtgtcgctg gtggtcggcg
     5881 gcaagctgtg gggcctggtt gtctgtcacc attatctgcc gcgcttcatc cgtttcgagc
     5941 tgcgggcgat ctgcaaacgg ctcgccgaaa ggatcgcgac gcggatcacc gcgcttgaga
     6001 gctaaaccgg ttagtaatga gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca
     6061 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     6121 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6181 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6241 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6301 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     6361 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6421 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6481 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6541 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6601 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6661 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6721 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     6781 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6841 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     6901 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     6961 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     7021 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     7081 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     7141 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     7201 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     7261 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     7321 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     7381 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     7441 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     7501 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     7561 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     7621 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     7681 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     7741 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     7801 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7861 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7921 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7981 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     8041 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     8101 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     8161 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     8221 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     8281 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     8341 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     8401 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     8461 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     8521 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     8581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     8641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     8701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     8941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     9001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     9061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     9121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     9181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     9241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     9301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     9361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     9421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     9481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     9541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
    10021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    10081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    10141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    10201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    10261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    10321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    10381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    10441 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
    10501 tggtgcactc tcagtacaat ctgctctgat gcc
//
