LOCUS       sequence-354107-       13678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing Cas9-VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13678)
  AUTHORS   Nghi Dang , Alissa Lance-Byrne, Angela Tung, Xiaoge Guo, Ryan J 
            Cecchi, Joanna Buchthal, Alejandro Chavez, Nan Cher Yeo
  TITLE     Generation and application of a versatile CRISPR toolkit for 
            mammalian cell engineering
  JOURNAL   Synthetic Biology, Volume 6, Issue 1, 2021
REFERENCE   2  (bases 1 to 13678)
  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: "Synthetic 
            Biology, Volume 6, Issue 1, 2021"
COMMENT     Sequence Label: NAX_CAGGS_CAS9_VPR_KanR neo
FEATURES             Location/Qualifiers
     source          1..13678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          111..1127
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1051..1073
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1135..1154
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1224..1233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1230..5333
                     /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             1230..5333
                     /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             1230..5333
                     /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             5346..5366
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    5403..5427
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5457..5606
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             5631..5651
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6081..6437
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             6456..7025
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     primer_bind     complement(7050..7069)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7094..7142
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7374..7585
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             7601..8395
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             7601..8395
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7655..7674)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8265..8284
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8399..8520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8436..8455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8490..8509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9283..9317
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9808..9824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9808..9824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9821..9843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9832..9848
                     /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(9856..9862,9863..9880,9881..9886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9901..9922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10039..10056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10210..10798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10290..10309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10969..11760,11761..11829))
                     /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     11592..11611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11830..11934)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11960..12415)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12097..12118)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12309..12328
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12542..12564
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12556..12573
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12557..12573
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   12703..12765
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        13186..13565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
       61 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      121 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      181 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      241 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
      301 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
      361 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
      421 gcgcggggct ttttgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
      481 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
      541 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
      601 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
      661 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
      721 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
      781 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
      841 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
      901 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
      961 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1021 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1081 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1141 tgctggttat tgtgctgtct catcattttg gcaaagaatc tagataccct cgtaaaggtc
     1201 tagagctaga ggatcgaacc cttgccacca tggacaagaa gtactccatt gggctcgata
     1261 tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa
     1321 aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc
     1381 tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca
     1441 gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg
     1501 ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata
     1561 aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa
     1621 agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact
     1681 tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg
     1741 agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga
     1801 cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa
     1861 tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg
     1921 gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca
     1981 actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct
     2041 acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt
     2101 tagcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg
     2161 agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag
     2221 acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt
     2281 tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg
     2341 aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg
     2401 taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc
     2461 cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc
     2521 cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact
     2581 atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag
     2641 agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct
     2701 tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac
     2761 actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca
     2821 cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc
     2881 tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa
     2941 agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc
     3001 tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg
     3061 agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga
     3121 tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc
     3181 tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc
     3241 gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc
     3301 ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag
     3361 cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc
     3421 cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa
     3481 tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc
     3541 agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac
     3601 tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc
     3661 tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca
     3721 atcggctctc cgactacgac gtggatcata tcgtgcccca gtcttttctc aaagatgatt
     3781 ctattgataa taaagtgttg acaagatccg ataaaaatag agggaagagt gataacgtcc
     3841 cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac
     3901 tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt
     3961 tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg
     4021 tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc
     4081 gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc
     4141 agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg
     4201 cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg
     4261 gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca
     4321 aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta
     4381 cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag
     4441 aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc
     4501 aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc
     4561 tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat
     4621 acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga
     4681 aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc
     4741 gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca
     4801 aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga
     4861 aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta
     4921 aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag
     4981 ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg
     5041 agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc
     5101 tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc
     5161 acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca
     5221 tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt
     5281 caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg
     5341 ctgaccccaa gaagaagagg aaggtgtcgc cagggatccg tcgacttgac gcgttaatat
     5401 caacaagttt gtacaaaaaa gcaggctaca aagaggccag cggttccgga cgggctgacg
     5461 cattggacga ttttgatctg gatatgctgg gaagtgacgc cctcgatgat tttgaccttg
     5521 acatgcttgg ttcggatgcc cttgatgact ttgacctcga catgctcggc agtgacgccc
     5581 ttgatgattt cgacctggac atgctgatta actctagaag ttccggatct ccgaaaaaga
     5641 aacgcaaagt tggtagccag tacctgcccg acaccgacga ccggcaccgg atcgaggaaa
     5701 agcggaagcg gacctacgag acattcaaga gcatcatgaa gaagtccccc ttcagcggcc
     5761 ccaccgaccc tagacctcca cctagaagaa tcgccgtgcc cagcagatcc agcgccagcg
     5821 tgccaaaacc tgccccccag ccttacccct tcaccagcag cctgagcacc atcaactacg
     5881 acgagttccc taccatggtg ttccccagcg gccagatctc tcaggcctct gctctggctc
     5941 cagcccctcc tcaggtgctg cctcaggctc ctgctcctgc accagctcca gccatggtgt
     6001 ctgcactggc tcaggcacca gcacccgtgc ctgtgctggc tcctggacct ccacaggctg
     6061 tggctccacc agcccctaaa cctacacagg ccggcgaggg cacactgtct gaagctctgc
     6121 tgcagctgca gttcgacgac gaggatctgg gagccctgct gggaaacagc accgatcctg
     6181 ccgtgttcac cgacctggcc agcgtggaca acagcgagtt ccagcagctg ctgaaccagg
     6241 gcatccctgt ggcccctcac accaccgagc ccatgctgat ggaatacccc gaggccatca
     6301 cccggctcgt gacaggcgct cagaggcctc ctgatccagc tcctgcccct ctgggagcac
     6361 caggcctgcc taatggactg ctgtctggcg acgaggactt cagctctatc gccgatatgg
     6421 atttctcagc cttgctgggc tctggcagcg gcagccggga ttccagggaa gggatgtttt
     6481 tgccgaagcc tgaggccggc tccgctatta gtgacgtgtt tgagggccgc gaggtgtgcc
     6541 agccaaaacg aatccggcca tttcatcctc caggaagtcc atgggccaac cgcccactcc
     6601 ccgccagcct cgcaccaaca ccaaccggtc cagtacatga gccagtcggg tcactgaccc
     6661 cggcaccagt ccctcagcca ctggatccag cgcccgcagt gactcccgag gccagtcacc
     6721 tgttggagga tcccgatgaa gagacgagcc aggctgtcaa agcccttcgg gagatggccg
     6781 atactgtgat tccccagaag gaagaggctg caatctgtgg ccaaatggac ctttcccatc
     6841 cgcccccaag gggccatctg gatgagctga caaccacact tgagtccatg accgaggatc
     6901 tgaacctgga ctcacccctg accccggaat tgaacgagat tctggatacc ttcctgaacg
     6961 acgagtgcct cttgcatgcc atgcatatca gcacaggact gtccatcttc gacacatctc
     7021 tgttttgagt ttaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     7081 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     7141 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     7201 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     7261 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgaaa
     7321 tcggatccgc ggccacactg attctagctg cagaggctcc ggtgcccgtc agtgggcaga
     7381 gcgcacatcg cccacagccc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
     7441 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     7501 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     7561 caacgggttt gccgccagaa cacaggatcc tcgagccacc atgattgaac aagatggatt
     7621 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     7681 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     7741 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     7801 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     7861 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     7921 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     7981 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     8041 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     8101 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     8161 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     8221 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     8281 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     8341 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagttaa
     8401 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8461 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8521 tcatgtctgg aattgactca aatgatgtca attagtctat cagaagctca tctggtctcc
     8581 cttccggggg acaagacatc cctgtttaat atttaaacag cagtgttccc aaactgggtt
     8641 cttatatccc ttgctctggt caaccaggtt gcagggtttc ctgtcctcac aggaacgaag
     8701 tccctaaaga aacagtggca gccaggttta gccccggaat tgactggatt ccttttttag
     8761 ggcccattgg tatggctttt tccccgtatc cccccaggtg tctgcaggct caaagagcag
     8821 cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg
     8881 cacgctgccg gctcggggat gcggggggag cgccggaccg gagcggagcc ccgggcggct
     8941 cgctgctgcc ccctagcggg ggagggacgt aattacatcc ctgggggctt tggggggggg
     9001 ctgtccctga tatctataac aagaaaatat atatataata agttatcacg taagtagaac
     9061 atgaaataac aatataatta tcgtatgagt taaatcttaa aagtcacgta aaagataatc
     9121 atgcgtcatt ttgactcacg cggtcgttat agttcaaaat cagtgacact taccgcattg
     9181 acaagcacgc ctcacgggag ctccaagcgg cgactgagat gtcctaaatg cacagcgacg
     9241 gattcgcgct atttagaaag agagagcaat atttcaagaa tgcatgcgtc aattttacgc
     9301 agactatctt tctagggtta atctagctgc atcaggatca tatcgtcggg tcttttttcc
     9361 ggctcagtca tcgcccaagc tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct
     9421 tttcccgcga ggttgaagcg gcatggaaag agtttgccga ggatgactgc tgctgcattg
     9481 acgttgagcg aaaacgcacg tttaccatga tgattcggga aggtgtggcc atgcacgcct
     9541 ttaacggtga actgttcgtt caggccacct gggataccag ttcgtcgcgg cttttccgga
     9601 cacagttccg gatggtcagc ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc
     9661 ggaactgccg tgccggtgtg cagattaatg acagcggtgc ggcgctggga tattacgtca
     9721 gcgaggacgg gtatcctggc tggatgccgc agaaatggac atggataccc cgtgagttac
     9781 ccggcgggcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     9841 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     9901 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     9961 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
    10021 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
    10081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
    10141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
    10201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
    10261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
    10321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
    10381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    10441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    10501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    10561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    10621 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    10681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    10741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    10801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
    10861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
    10921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
    10981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
    11041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
    11101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
    11161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
    11221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
    11281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
    11341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
    11401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
    11461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
    11521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
    11581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
    11641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
    11701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
    11761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    11821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
    11881 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
    11941 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
    12001 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
    12061 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
    12121 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
    12181 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
    12241 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
    12301 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
    12361 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
    12421 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
    12481 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
    12541 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cctcgttcat tcacgttttt
    12601 gaacccgtgg aggacgggca gactcgcggt gcaaatgtgt tttacagcgt gatggagcag
    12661 atgaagatgc tcgacacgct gcagaacacg cagctagatt aaccctagaa agataatcat
    12721 attgtgacgt acgttaaaga taatcatgtg taaaattgac gcatgtgttt tatcggtctg
    12781 tatatcgagg tttatttatt aatttgaata gatattaagt tttattatat ttacacttac
    12841 atactaataa taaattcaac aaacaattta tttatgttta tttatttatt aaaaaaaaca
    12901 aaaactcaaa atttcttcta taaagtaaca aaacttttat gagggacagc ccccccccaa
    12961 agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg
    13021 gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc
    13081 cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga
    13141 gcctgcagac acctgggggg atacggggaa aaggcctcca cggccgacat tgattattga
    13201 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
    13261 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
    13321 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
    13381 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
    13441 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
    13501 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
    13561 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
    13621 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggg
//
