LOCUS       sequence-371891-       14514 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing Cas9-VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 14514)
  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 14514)
  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_mcherry
FEATURES             Location/Qualifiers
     source          1..14514
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          671..1688
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1612..1634
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1696..1715
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1785..1794
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1791..5894
                     /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             1791..5894
                     /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             1791..5894
                     /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             5907..5927
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    5964..5988
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6018..6167
                     /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             6192..6212
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6642..6998
                     /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             7017..7586
                     /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(7611..7630)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7655..7703
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7935..8146
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8156..8165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(8280..8298)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8548..8567
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    8849..8970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8886..8905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8940..8959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9733..9767
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10258..10274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10258..10274)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10271..10293)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10282..10298
                     /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(10306..10312,10313..10330,10331..10336))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10351..10372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10489..10506)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10660..11248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10740..10759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11419..12210,12211..12279))
                     /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     12042..12061
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12280..12384)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(12410..12865)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12547..12568)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12759..12778
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12992..13014
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13006..13023
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13007..13023
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   13153..13215
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        13636..14015
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 scsscscsms scsscsscss mscsssscmc sssscsmmss cmscscscms csmcycccsc
       61 ccsccscscs cyycscyyyy ymymsssccs ccsccsccsc csccccsccc ymmmmssmmm
      121 cyyycssmsc scsccscccy smyysscysc csccsccccc cccccccccs cccccccccc
      181 cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc
      241 cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc
      301 cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc
      361 cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc
      421 cccccccccc cccccccccc cccccccccc aattttgttk tttttttttt ttttadwtkk
      481 ttgggggggg gggggggggg gggggggggg ggggcgcgcg csrggsgggg sggggcgggg
      541 cgrggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
      601 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
      661 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
      721 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      781 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
      841 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
      901 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
      961 ctgtgagcgc tgcgggcgcg gcgcggggct ttttgcgctc cgcagtgtgc gcgaggggag
     1021 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     1081 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     1141 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     1201 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg kggcggcagg kgggggkgcc
     1261 gggcggggcg gggcncgccy cgggccgggg rgggctcggg ggaggggcgc ggcggccccc
     1321 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
     1381 gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc
     1441 gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa
     1501 tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc
     1561 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
     1621 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     1681 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     1741 ctagataccc tcgtaaaggt ctagagctag cggatcgaac ccttgccacc atggacaaga
     1801 agtactccat tgggctcgat atcggcacaa acagcgtcgg ctgggccgtc attacggacg
     1861 agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc cacagcataa
     1921 agaagaacct cattggcgcc ctcctgttcg actccgggga gacggccgaa gccacgcggc
     1981 tcaaaagaac agcacggcgc agatataccc gcagaaagaa tcggatctgc tacctgcagg
     2041 agatctttag taatgagatg gctaaggtgg atgactcttt cttccatagg ctggaggagt
     2101 cctttttggt ggaggaggat aaaaagcacg agcgccaccc aatctttggc aatatcgtgg
     2161 acgaggtggc gtaccatgaa aagtacccaa ccatatatca tctgaggaag aagcttgtag
     2221 acagtactga taaggctgac ttgcggttga tctatctcgc gctggcgcat atgatcaaat
     2281 ttcggggaca cttcctcatc gagggggacc tgaacccaga caacagcgat gtcgacaaac
     2341 tctttatcca actggttcag acttacaatc agcttttcga agagaacccg atcaacgcat
     2401 ccggagttga cgccaaagca atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa
     2461 acctcatcgc acagctccct ggggagaaga agaacggcct gtttggtaat cttatcgccc
     2521 tgtcactcgg gctgaccccc aactttaaat ctaacttcga cctggccgaa gatgccaagc
     2581 ttcaactgag caaagacacc tacgatgatg atctcgacaa tctgctggcc cagatcggcg
     2641 accagtacgc agaccttttt ttagcggcaa agaacctgtc agacgccatt ctgctgagtg
     2701 atattctgcg agtgaacacg gagatcacca aagctccgct gagcgctagt atgatcaagc
     2761 gctatgatga gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga cagcaactgc
     2821 ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc ggatacattg
     2881 acggcggagc aagccaggag gaattttaca aatttattaa gcccatcttg gaaaaaatgg
     2941 acggcaccga ggagctgctg gtaaagctta acagagaaga tctgttgcgc aaacagcgca
     3001 ctttcgacaa tggaagcatc ccccaccaga ttcacctggg cgaactgcac gctatcctca
     3061 ggcggcaaga ggatttctac ccctttttga aagataacag ggaaaagatt gagaaaatcc
     3121 tcacatttcg gataccctac tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt
     3181 ggatgactcg caaatcagaa gagaccatca ctccctggaa cttcgaggaa gtcgtggata
     3241 agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa aatctgccta
     3301 acgaaaaggt gcttcctaaa cactctctgc tgtacgagta cttcacagtt tataacgagc
     3361 tcaccaaggt caaatacgtc acagaaggga tgagaaagcc agcattcctg tctggagagc
     3421 agaagaaagc tatcgtggac ctcctcttca agacgaaccg gaaagttacc gtgaaacagc
     3481 tcaaagaaga ctatttcaaa aagattgaat gtttcgactc tgttgaaatc agcggagtgg
     3541 aggatcgctt caacgcatcc ctgggaacgt atcacgatct cctgaaaatc attaaagaca
     3601 aggacttcct ggacaatgag gagaacgagg acattcttga ggacattgtc ctcaccctta
     3661 cgttgtttga agatagggag atgattgaag aacgcttgaa aacttacgct catctcttcg
     3721 acgacaaagt catgaaacag ctcaagaggc gccgatatac aggatggggg cggctgtcaa
     3781 gaaaactgat caatgggatc cgagacaagc agagtggaaa gacaatcctg gattttctta
     3841 agtccgatgg atttgccaac cggaacttca tgcagttgat ccatgatgac tctctcacct
     3901 ttaaggagga catccagaaa gcacaagttt ctggccaggg ggacagtctt cacgagcaca
     3961 tcgctaatct tgcaggtagc ccagctatca aaaagggaat actgcagacc gttaaggtcg
     4021 tggatgaact cgtcaaagta atgggaaggc ataagcccga gaatatcgtt atcgagatgg
     4081 cccgagagaa ccaaactacc cagaagggac agaagaacag tagggaaagg atgaagagga
     4141 ttgaagaggg tataaaagaa ctggggtccc aaatccttaa ggaacaccca gttgaaaaca
     4201 cccagcttca gaatgagaag ctctacctgt actacctgca gaacggcagg gacatgtacg
     4261 tggatcagga actggacatc aatcggctct ccgactacga cgtggatcat atcgtgcccc
     4321 agtcttttct caaagatgat tctattgata ataaagtgtt gacaagatcc gataaaaata
     4381 gagggaagag tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa aattattggc
     4441 ggcagctgct gaacgccaaa ctgatcacac aacggaagtt cgataatctg actaaggctg
     4501 aacgaggtgg cctgtctgag ttggataaag ccggcttcat caaaaggcag cttgttgaga
     4561 cacgccagat caccaagcac gtggcccaaa ttctcgattc acgcatgaac accaagtacg
     4621 atgaaaatga caaactgatt cgagaggtga aagttattac tctgaagtct aagctggtct
     4681 cagatttcag aaaggacttt cagttttata aggtgagaga gatcaacaat taccaccatg
     4741 cgcatgatgc ctacctgaat gcagtggtag gcactgcact tatcaaaaaa tatcccaagc
     4801 ttgaatctga atttgtttac ggagactata aagtgtacga tgttaggaaa atgatcgcaa
     4861 agtctgagca ggaaataggc aaggccaccg ctaagtactt cttttacagc aatattatga
     4921 attttttcaa gaccgagatt acactggcca atggagagat tcggaagcga ccacttatcg
     4981 aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc
     5041 ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta cagaccggag
     5101 gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa
     5161 aagattggga ccccaagaaa tacggcggat tcgattctcc tacagtcgct tacagtgtac
     5221 tggttgtggc caaagtggag aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc
     5281 tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg
     5341 cgaaaggata taaagaggtc aaaaaagacc tcatcattaa gcttcccaag tactctctct
     5401 ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta
     5461 acgagctggc actgccctct aaatacgtta atttcttgta tctggccagc cactatgaaa
     5521 agctcaaagg gtctcccgaa gataatgagc agaagcagct gttcgtggaa caacacaaac
     5581 actaccttga tgagatcatc gagcaaataa gcgaattctc caaaagagtg atcctcgccg
     5641 acgctaacct cgataaggtg ctttctgctt acaataagca cagggataag cccatcaggg
     5701 agcaggcaga aaacattatc cacttgttta ctctgaccaa cttgggcgcg cctgcagcct
     5761 tcaagtactt cgacaccacc atagacagaa agcggtacac ctctacaaag gaggtcctgg
     5821 acgccacact gattcatcag tcaattacgg ggctctatga aacaagaatc gacctctctc
     5881 agctcggtgg agacagcagg gctgacccca agaagaagag gaaggtgtcg ccagggatcc
     5941 gtcgacttga cgcgttaata tcaacaagtt tgtacaaaaa agcaggctac aaagaggcca
     6001 gcggttccgg acgggctgac gcattggacg attttgatct ggatatgctg ggaagtgacg
     6061 ccctcgatga ttttgacctt gacatgcttg gttcggatgc ccttgatgac tttgacctcg
     6121 acatgctcgg cagtgacgcc cttgatgatt tcgacctgga catgctgatt aactctagaa
     6181 gttccggatc tccgaaaaag aaacgcaaag ttggtagcca gtacctgccc gacaccgacg
     6241 accggcaccg gatcgaggaa aagcggaagc ggacctacga gacattcaag agcatcatga
     6301 agaagtcccc cttcagcggc cccaccgacc ctagacctcc acctagaaga atcgccgtgc
     6361 ccagcagatc cagcgccagc gtgccaaaac ctgcccccca gccttacccc ttcaccagca
     6421 gcctgagcac catcaactac gacgagttcc ctaccatggt gttccccagc ggccagatct
     6481 ctcaggcctc tgctctggct ccagcccctc ctcaggtgct gcctcaggct cctgctcctg
     6541 caccagctcc agccatggtg tctgcactgg ctcaggcacc agcacccgtg cctgtgctgg
     6601 ctcctggacc tccacaggct gtggctccac cagcccctaa acctacacag gccggcgagg
     6661 gcacactgtc tgaagctctg ctgcagctgc agttcgacga cgaggatctg ggagccctgc
     6721 tgggaaacag caccgatcct gccgtgttca ccgacctggc cagcgtggac aacagcgagt
     6781 tccagcagct gctgaaccag ggcatccctg tggcccctca caccaccgag cccatgctga
     6841 tggaataccc cgaggccatc acccggctcg tgacaggcgc tcagaggcct cctgatccag
     6901 ctcctgcccc tctgggagca ccaggcctgc ctaatggact gctgtctggc gacgaggact
     6961 tcagctctat cgccgatatg gatttctcag ccttgctggg ctctggcagc ggcagccggg
     7021 attccaggga agggatgttt ttgccgaagc ctgaggccgg ctccgctatt agtgacgtgt
     7081 ttgagggccg cgaggtgtgc cagccaaaac gaatccggcc atttcatcct ccaggaagtc
     7141 catgggccaa ccgcccactc cccgccagcc tcgcaccaac accaaccggt ccagtacatg
     7201 agccagtcgg gtcactgacc ccggcaccag tccctcagcc actggatcca gcgcccgcag
     7261 tgactcccga ggccagtcac ctgttggagg atcccgatga agagacgagc caggctgtca
     7321 aagcccttcg ggagatggcc gatactgtga ttccccagaa ggaagaggct gcaatctgtg
     7381 gccaaatgga cctttcccat ccgcccccaa ggggccatct ggatgagctg acaaccacac
     7441 ttgagtccat gaccgaggat ctgaacctgg actcacccct gaccccggaa ttgaacgaga
     7501 ttctggatac cttcctgaac gacgagtgcc tcttgcatgc catgcatatc agcacaggac
     7561 tgtccatctt cgacacatct ctgttttgag tttaaacggg ggaggctaac tgaaacacgg
     7621 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     7681 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     7741 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     7801 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aatgtcgggg cggcaggccc
     7861 tgccatagca gatctgcgaa atcggatccg cggccgcact gattctagct gcagaggctc
     7921 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     7981 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     8041 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     8101 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggatc ctcgagccac
     8161 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     8221 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     8281 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     8341 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     8401 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     8461 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     8521 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     8581 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     8641 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     8701 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     8761 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     8821 cggcatggac gagctgtaca agtaagttaa cttgtttatt gcagcttata atggttacaa
     8881 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     8941 tggtttgtcc aaactcatca atgtatctta tcatgtctgg aattgactca aatgatgtca
     9001 attagtctat cagaagctca tctggtctcc cttccggggg acaagacatc cctgtttaat
     9061 atttaaacag cagtgttccc aaactgggtt cttatatccc ttgctctggt caaccaggtt
     9121 gcagggtttc ctgtcctcac aggaacgaag tccctaaaga aacagtggca gccaggttta
     9181 gccccggaat tgactggatt ccttttttag ggcccattgg tatggctttt tccccgtatc
     9241 cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt
     9301 gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag
     9361 cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt
     9421 aattacatcc ctgggggctt tggggggggg ctgtccctga tatctataac aagaaaatat
     9481 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     9541 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     9601 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
     9661 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
     9721 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc
     9781 atcaggatca tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc
     9841 tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag
     9901 agtttgccga ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga
     9961 tgattcggga aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct
    10021 gggataccag ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca
    10081 tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg
    10141 acagcggtgc ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc
    10201 agaaatggac atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc
    10261 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    10321 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    10381 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    10441 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    10501 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    10561 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    10621 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    10681 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    10741 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    10801 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    10861 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    10921 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    10981 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    11041 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    11101 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    11161 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    11221 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    11281 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    11341 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    11401 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    11461 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    11521 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    11581 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    11641 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    11701 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    11761 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    11821 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    11881 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    11941 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    12001 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    12061 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    12121 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    12181 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    12241 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    12301 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    12361 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
    12421 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
    12481 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
    12541 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
    12601 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
    12661 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
    12721 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
    12781 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
    12841 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
    12901 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
    12961 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
    13021 agtgagcgcg cctcgttcat tcacgttttt gaacccgtgg aggacgggca gactcgcggt
    13081 gcaaatgtgt tttacagcgt gatggagcag atgaagatgc tcgacacgct gcagaacacg
    13141 cagctagatt aaccctagaa agataatcat attgtgacgt acgttaaaga taatcatgtg
    13201 taaaattgac gcatgtgttt tatcggtctg tatatcgagg tttatttatt aatttgaata
    13261 gatattaagt tttattatat ttacacttac atactaataa taaattcaac aaacaattta
    13321 tttatgttta tttatttatt aaaaaaaaca aaaactcaaa atttcttcta taaagtaaca
    13381 aaacttttat gagggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc
    13441 ccgctagggg gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca
    13501 tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct
    13561 ttcctctgaa cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa
    13621 aaggcctcca cggccgacat tgattattga ctagttatta atagtaatca attacggggt
    13681 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
    13741 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
    13801 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
    13861 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
    13921 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
    13981 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
    14041 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
    14101 tattttgtgc agcgatgggg gcgggggggg ggggggssgn gcsssnggng gggggggggg
    14161 gggggggggg gggggggggg gggggggggg gggggggggg gggggssrrk nngggggggg
    14221 gggggggsgg grgggggggg gggggggggg gggggggggg gggggggggg gggggggggg
    14281 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
    14341 gggggggggg gggggggggn gggggggggk gggggggggg gggggggggg ggggrgnngg
    14401 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
    14461 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgc
//
