LOCUS       sequence-354045-       11316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing spCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11316)
  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 11316)
  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_KanR neo
FEATURES             Location/Qualifiers
     source          1..11316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     309..331
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     393..412
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      481..490
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             487..4590
                     /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             487..4590
                     /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             487..4590
                     /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             4603..4623
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4678..4697)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4722..4770
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5002..5213
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5229..6023
                     /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             5229..6023
                     /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(5283..5302)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5893..5912
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6027..6148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6064..6083)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6118..6137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6911..6945
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7436..7452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7436..7452)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7449..7471)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7460..7476
                     /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(7484..7490,7491..7508,7509..7514))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7529..7550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7667..7684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7838..8426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7918..7937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8597..9388,9389..9457))
                     /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     9220..9239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9458..9562)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9588..10043)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9725..9746)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9937..9956
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10170..10192
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10184..10201
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10185..10201
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   10331..10393
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        10814..11193
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
       61 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
      121 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
      181 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
      241 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
      301 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
      361 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
      421 tcattttggc aaagaatcta gataccctcg taaaggtcta gagctagcga attcgaattt
      481 gccaccatgg acaagaagta ctccattggg ctcgatatcg gcacaaacag cgtcggctgg
      541 gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc
      601 gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg
      661 gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg
      721 atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc
      781 cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc
      841 tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg
      901 aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg
      961 gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac
     1021 agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag
     1081 aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa
     1141 tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt
     1201 ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg
     1261 gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg
     1321 ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac
     1381 gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc
     1441 gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt
     1501 gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc
     1561 tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc
     1621 atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg
     1681 ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa
     1741 ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa
     1801 aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga
     1861 aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc
     1921 gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt
     1981 gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc
     2041 acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca
     2101 ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa
     2161 gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt
     2221 gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg
     2281 aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac
     2341 attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact
     2401 tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga
     2461 tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca
     2521 atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat
     2581 gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac
     2641 agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg
     2701 cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat
     2761 atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg
     2821 gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa
     2881 cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac
     2941 ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg
     3001 gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca
     3061 agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa
     3121 atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat
     3181 aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa
     3241 aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc
     3301 atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg
     3361 aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc
     3421 aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc
     3481 aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt
     3541 aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt
     3601 tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg
     3661 aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg
     3721 gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc
     3781 gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag
     3841 ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca
     3901 gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa
     3961 agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc
     4021 atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt
     4081 cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc
     4141 gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg
     4201 gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc
     4261 gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa
     4321 agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg
     4381 gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg
     4441 ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct
     4501 acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca
     4561 agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag
     4621 gtgtgaaagg gttcgatccc taccggttag taatgagttt aaacggggga ggctaactga
     4681 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4741 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4801 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4861 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4921 caggccctgc catagcagat ctgcgaaatc ggatccgcgg ccacactgat tctagctgca
     4981 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagccccc gagaagttgg
     5041 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     5101 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     5161 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggatcctc
     5221 gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5281 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5341 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5401 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5461 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5521 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5581 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5641 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5701 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5761 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5821 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5881 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5941 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     6001 gccttcttga cgagttcttc tgagttaact tgtttattgc agcttataat ggttacaaat
     6061 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6121 gtttgtccaa actcatcaat gtatcttatc atgtctggaa ttgactcaaa tgatgtcaat
     6181 tagtctatca gaagctcatc tggtctccct tccgggggac aagacatccc tgtttaatat
     6241 ttaaacagca gtgttcccaa actgggttct tatatccctt gctctggtca accaggttgc
     6301 agggtttcct gtcctcacag gaacgaagtc cctaaagaaa cagtggcagc caggtttagc
     6361 cccggaattg actggattcc ttttttaggg cccattggta tggctttttc cccgtatccc
     6421 cccaggtgtc tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc
     6481 caccttcccc gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg
     6541 ccggaccgga gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa
     6601 ttacatccct gggggctttg ggggggggct gtccctgata tctataacaa gaaaatatat
     6661 atataataag ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta
     6721 aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag
     6781 ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg
     6841 actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat
     6901 ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat
     6961 caggatcata tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg gcgctatctg
     7021 ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc atggaaagag
     7081 tttgccgagg atgactgctg ctgcattgac gttgagcgaa aacgcacgtt taccatgatg
     7141 attcgggaag gtgtggccat gcacgccttt aacggtgaac tgttcgttca ggccacctgg
     7201 gataccagtt cgtcgcggct tttccggaca cagttccgga tggtcagccc gaagcgcatc
     7261 agcaacccga acaataccgg cgacagccgg aactgccgtg ccggtgtgca gattaatgac
     7321 agcggtgcgg cgctgggata ttacgtcagc gaggacgggt atcctggctg gatgccgcag
     7381 aaatggacat ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa tcatggtcat
     7441 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7501 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7561 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7621 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7681 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7741 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7801 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7861 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7921 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7981 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     8041 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     8101 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     8161 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     8221 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     8281 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8341 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     8401 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8461 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8521 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8581 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8641 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8701 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8761 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8821 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8881 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8941 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     9001 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     9061 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9121 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9181 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     9241 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9301 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9361 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9421 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9481 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9541 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     9601 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     9661 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     9721 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     9781 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     9841 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     9901 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     9961 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
    10021 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
    10081 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
    10141 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
    10201 tgagcgcgcc tcgttcattc acgtttttga acccgtggag gacgggcaga ctcgcggtgc
    10261 aaatgtgttt tacagcgtga tggagcagat gaagatgctc gacacgctgc agaacacgca
    10321 gctagattaa ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgtgta
    10381 aaattgacgc atgtgtttta tcggtctgta tatcgaggtt tatttattaa tttgaataga
    10441 tattaagttt tattatattt acacttacat actaataata aattcaacaa acaatttatt
    10501 tatgtttatt tatttattaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa
    10561 acttttatga gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc
    10621 gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc
    10681 cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt
    10741 cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa
    10801 ggcctccacg gccgacattg attattgact agttattaat agtaatcaat tacggggtca
    10861 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
    10921 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
    10981 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
    11041 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
    11101 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
    11161 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
    11221 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
    11281 ttttgtgcag cgatgggggc gggggggggg gggggg
//
