LOCUS       sequence-363635-        9609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV and T7 promoter expression plasmid for human codon usage 
            anCas(Spyo_Cas9) with an N-terminal NLS and C-terminal NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9609)
  AUTHORS   Alonso-Lerma B, Jabalera Y, Samperio S, Morin M, Fernandez A, Hille 
            LT, Silverstein RA, Quesada-Ganuza A, Reifs A, Fernandez-Penalver S,
            Benitez Y, Soletto L, Gavira JA, Diaz A, Vranken W, Sanchez-Mejias 
            A, Guell M, Mojica FJM, Kleinstiver BP, Moreno-Pelayo MA, Montoliu 
            L, Perez-Jimenez R
  TITLE     Evolution of CRISPR-associated endonucleases as inferred from 
            resurrected proteins.
  JOURNAL   Nat Microbiol. 2023 Jan;8(1):77-90. doi: 10.1038/s41564-022-01265-y.
            Epub 2023 Jan 2.
  PUBMED    36593295
REFERENCE   2  (bases 1 to 9609)
  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/s41564-022-01265-y"; journalName: "Nat Microbiol"; date: 
            "2023-01"; volume: "8"; issue: "1"; pages: "77-90"
COMMENT     Sequence Label: pCMV-T7-NLS(SV40)-(Spyo_Cas9)-6xHis-NLS(SV40)
FEATURES             Location/Qualifiers
     source          1..9609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..72)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        244..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        624..827
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     778..798
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     872..891
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        872..890
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             913..933
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             940..5043
                     /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             940..5043
                     /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             5044..5061
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5080..5100
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5209..5226)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5215..5439
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5485..5913
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5572..5591)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5782..5803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5922..5942)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5927..6256
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6107..6242
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6169..6188
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6323..7117
                     /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(6377..6396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6987..7006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7291..7412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7328..7347)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7382..7401
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7461..7477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7461..7477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7474..7496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7485..7501
                     /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(7509..7515,7516..7533,7534..7539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7554..7575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7692..7709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7863..8451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7943..7962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8622..9413,9414..9482))
                     /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     9245..9264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9483..9587)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
       61 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
      121 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
      181 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
      241 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      661 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      721 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      781 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      841 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      901 ctggctagca tgcccaagaa aaagcggaag gtgcccaaga tggacaagaa gtacagcatc
      961 ggcctggaca tcggcaccaa ttctgttggc tgggccgtga tcaccgacga gtacaaggtg
     1021 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaatctg
     1081 atcggcgccc tgctgttcga ctctggcgaa acagccgaag ccaccagact gaagagaacc
     1141 gccagacggc ggtacaccag aagaaagaac cggatctgct acctgcaaga gatcttcagc
     1201 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     1261 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga tgaggtggcc
     1321 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     1381 aaggccgacc tgagactgat ctatctggcc ctggctcaca tgatcaagtt ccggggccac
     1441 ttcctgatcg agggcgacct gaatcctgac aacagcgacg tggacaagct gttcatccag
     1501 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggagtggat
     1561 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa cctgatcgct
     1621 cagctgcccg gcgagaagaa gaatggcctg ttcggcaacc tgattgccct gagcctgggc
     1681 ctgacaccta acttcaagag caacttcgac ctggccgagg acgccaaact gcagctgtcc
     1741 aaggacacct acgacgacga cctggacaat ctgctggccc agatcggcga tcagtacgcc
     1801 gacttgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     1861 gtgaacaccg agatcacaaa ggcccctctg agcgcctcta tgatcaagag atacgacgag
     1921 caccaccagg atctgaccct gctgaaggcc ctcgttagac agcagctgcc tgagaagtac
     1981 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga tggcggagcc
     2041 agccaagagg aattctacaa gttcatcaag cccatcctcg agaagatgga cggcaccgag
     2101 gaactgctgg tcaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaat
     2161 ggctctatcc ctcaccagat ccacctggga gagctgcacg ccattctgcg gagacaagag
     2221 gacttttacc cattcctgaa ggacaaccgg gaaaagattg agaagatcct gaccttcagg
     2281 atcccctact acgtgggacc actggccaga ggcaatagca gattcgcctg gatgaccaga
     2341 aagagcgagg aaaccatcac accctggaac ttcgaggaag tggtggacaa gggcgccagc
     2401 gctcagtcct tcatcgagcg gatgaccaac ttcgataaga acctgcctaa cgagaaggtg
     2461 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt acaacgagct gaccaaagtg
     2521 aaatacgtga ccgagggaat gagaaagccc gcctttctga gcggcgagca gaaaaaggcc
     2581 attgtggatc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     2641 tacttcaaga aaatcgagtg cttcgacagc gtggaaatca gcggcgtgga agatcggttc
     2701 aatgccagcc tgggcacata ccacgacctg ctgaaaatta tcaaggacaa ggacttcctg
     2761 gacaacgaag agaacgagga catcctggaa gatatcgtgc tgaccctgac actgtttgag
     2821 gacagagaga tgatcgagga acggctgaaa acatacgccc acctgttcga cgacaaagtg
     2881 atgaagcaac tgaagcggcg gagatacacc ggctggggca gactgtctcg gaagctgatc
     2941 aacggcatcc gggataagca gtccggcaag accatcctgg actttctgaa gtccgacggc
     3001 ttcgccaatc ggaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggat
     3061 atccagaaag cccaggtgtc cggccagggc gattctctgc atgagcacat tgccaacctg
     3121 gccggctctc ccgccattaa gaagggcatt ctgcagacag tgaaggtggt ggacgagctg
     3181 gtcaaagtca tgggcagaca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     3241 cagaccacac agaagggcca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     3301 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     3361 aacgagaagc tgtacctgta ctacctccag aacggccggg atatgtacgt ggaccaagag
     3421 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcccca gtcttttctg
     3481 aaggacgact ccatcgacaa caaggtcctg accagaagcg acaagaaccg gggcaagagc
     3541 gataacgtgc cctccgaaga ggtggtcaag aagatgaaga actactggcg acagctgctg
     3601 aacgccaagc tgattaccca gcggaagttc gataacctga ccaaggccga gagaggcggc
     3661 ctgtctgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     3721 accaaacacg tggcacagat tctggactcc cggatgaaca ctaagtacga cgagaatgac
     3781 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     3841 aaggatttcc agttctacaa agtgcgcgag atcaacaact accatcacgc ccacgacgcc
     3901 tacctgaatg ccgttgttgg aacagccctg atcaagaagt atcccaagct ggaaagcgag
     3961 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcaa
     4021 gagattggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     4081 accgagatta ccctggccaa cggcgagatc agaaagcggc ctctgatcga gacaaacggc
     4141 gaaaccggcg agattgtgtg ggataagggc agagactttg ccacagtgcg gaaagtgctg
     4201 agcatgcccc aagtgaatat cgtgaagaaa accgaggtgc agacaggcgg cttcagcaaa
     4261 gagtccattc tgcccaagag aaacagcgat aagctgatcg cccggaagaa ggactgggac
     4321 cctaagaagt acggcggctt cgatagccct accgtggcct attctgtgct ggtggtggcc
     4381 aaagtggaaa agggcaagtc caagaaactc aagagcgtga aagagctgct ggggatcacc
     4441 atcatggaaa gaagcagctt cgagaagaat cctatcgatt tcctcgaggc caagggctac
     4501 aaagaagtga aaaaggacct gatcatcaag ctccccaagt actccctgtt cgagctggaa
     4561 aatggccgga agcggatgct ggcttctgct ggcgaactgc agaagggaaa cgaactggcc
     4621 ctgcctagca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     4681 agccccgagg acaatgagca aaagcagctg tttgtggaac agcacaagca ctacctggac
     4741 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     4801 gacaaagtgc tgtccgccta caacaagcac cgggacaagc ctatcagaga gcaggccgag
     4861 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     4921 gacaccacca tcgaccggaa gcgctacacc agcaccaaag aggtgctgga cgccacactg
     4981 atccaccagt ctatcaccgg cctgtacgag acacggatcg acctgtctca gctcggaggt
     5041 gatcaccatc atcatcacca cgcctctgga acaggcggac ctaaaaagaa gaggaaggtt
     5101 taaggtaccg agctcggatc cactagtcca gtgtggtgga attctgcaga tatccagcac
     5161 agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     5221 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     5281 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     5341 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     5401 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     5461 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5521 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5581 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5641 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5701 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5761 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5821 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5881 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     5941 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6001 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6061 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     6121 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6181 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6241 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     6301 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     6361 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     6421 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     6481 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     6541 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     6601 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6661 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6721 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6781 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6841 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6901 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6961 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     7021 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     7081 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     7141 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     7201 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     7261 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     7321 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7381 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     7441 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7501 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7561 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7621 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7681 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7741 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7801 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7861 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7921 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7981 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     8041 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     8101 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     8161 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     8221 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8281 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     8341 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8401 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8461 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8521 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8581 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8641 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8701 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8761 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8821 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8881 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8941 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     9001 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     9061 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     9121 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     9181 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     9241 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9301 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9361 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9421 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9481 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9541 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9601 aaagtgcca
//
