LOCUS       sequence-352732-       11706 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 11706)
  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 11706)
  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_Blasto
FEATURES             Location/Qualifiers
     source          1..11706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..657
                     /label=chicken beta-actin promoter
     intron          658..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1770..1779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1776..5879
                     /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             1776..5879
                     /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             1776..5879
                     /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             5892..5912
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5967..5986)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6011..6059
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6291..6502
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6512..6521
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6518..6916
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6920..7041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6957..6976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7011..7030
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7804..7838
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8329..8345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8329..8345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8342..8364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8353..8369
                     /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(8377..8383,8384..8401,8402..8407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8422..8443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8560..8577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8731..9319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8811..8830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9490..10281,10282..10350))
                     /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     10113..10132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10351..10455)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10481..10936)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10618..10639)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10830..10849
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11063..11085
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11077..11094
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11078..11094
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11224..11286
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgccr ggsggggsgg ggsggggsgr ggggsggggs
      541 ggggsgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      661 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      721 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      781 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      841 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
      901 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      961 gggcgcggcg cggggctttt tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1081 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaatctag ataccctcgt
     1741 aaaggtctag agctagcgaa ttcgaatttg ccaccatgga caagaagtac tccattgggc
     1801 tcgatatcgg cacaaacagc gtcggctggg ccgtcattac ggacgagtac aaggtgccga
     1861 gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag aacctcattg
     1921 gcgccctcct gttcgactcc ggggagacgg ccgaagccac gcggctcaaa agaacagcac
     1981 ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc tttagtaatg
     2041 agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt ttggtggagg
     2101 aggataaaaa gcacgagcgc cacccaatct ttggcaatat cgtggacgag gtggcgtacc
     2161 atgaaaagta cccaaccata tatcatctga ggaagaagct tgtagacagt actgataagg
     2221 ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc
     2281 tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt atccaactgg
     2341 ttcagactta caatcagctt ttcgaagaga acccgatcaa cgcatccgga gttgacgcca
     2401 aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc atcgcacagc
     2461 tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca ctcgggctga
     2521 cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag
     2581 acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag tacgcagacc
     2641 tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt ctgcgagtga
     2701 acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat gatgagcacc
     2761 accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag aagtacaagg
     2821 aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc ggagcaagcc
     2881 aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc accgaggagc
     2941 tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa
     3001 gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg caagaggatt
     3061 tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca tttcggatac
     3121 cctactatgt aggccccctc gcccggggaa attccagatt cgcgtggatg actcgcaaat
     3181 cagaagagac catcactccc tggaacttcg aggaagtcgt ggataagggg gcctctgccc
     3241 agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa aaggtgcttc
     3301 ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc aaggtcaaat
     3361 acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag aaagctatcg
     3421 tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa gaagactatt
     3481 tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat cgcttcaacg
     3541 catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac ttcctggaca
     3601 atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg tttgaagata
     3661 gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac aaagtcatga
     3721 aacagctcaa gaggcgccga tatacaggat gggggcggct gtcaagaaaa ctgatcaatg
     3781 ggatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc gatggatttg
     3841 ccaaccggaa cttcatgcag ttgatccatg atgactctct cacctttaag gaggacatcc
     3901 agaaagcaca agtttctggc cagggggaca gtcttcacga gcacatcgct aatcttgcag
     3961 gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat gaactcgtca
     4021 aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga gagaaccaaa
     4081 ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa gagggtataa
     4141 aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag cttcagaatg
     4201 agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat caggaactgg
     4261 acatcaatcg gctctccgac tacgacgtgg atcatatcgt gccccagtct tttctcaaag
     4321 atgattctat tgataataaa gtgttgacaa gatccgataa aaatagaggg aagagtgata
     4381 acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg
     4441 ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga ggtggcctgt
     4501 ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca
     4561 agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa aatgacaaac
     4621 tgattcgaga ggtgaaagtt attactctga agtctaagct ggtctcagat ttcagaaagg
     4681 actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat gatgcctacc
     4741 tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagcttgaa tctgaatttg
     4801 tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa
     4861 taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt ttcaagaccg
     4921 agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca aacggagaaa
     4981 caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccggaag gtcctgtcca
     5041 tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc tccaaggaaa
     5101 gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat tgggacccca
     5161 agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt gtggccaaag
     5221 tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc atcacaatca
     5281 tggagcgatc aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa ggatataaag
     5341 aggtcaaaaa agacctcatc attaagcttc ccaagtactc tctctttgag cttgaaaacg
     5401 gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag ctggcactgc
     5461 cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc aaagggtctc
     5521 ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac cttgatgaga
     5581 tcatcgagca aataagcgaa ttctccaaaa gagtgatcct cgccgacgct aacctcgata
     5641 aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag gcagaaaaca
     5701 ttatccactt gtttactctg accaacttgg gcgcgcctgc agccttcaag tacttcgaca
     5761 ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc acactgattc
     5821 atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc ggtggagaca
     5881 gcagggctga ccccaagaag aagaggaagg tgtgaaaggg ttcgatccct accggttagt
     5941 aatgagttta aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     6001 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     6061 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     6121 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     6181 cccagggctc gcagccaatg tcggggcggc aggccctgcc atagcagatc tgcgaaatcg
     6241 gatccgcggc cgcactgatt ctagctgcag aggctccggt gcccgtcagt gggcagagcg
     6301 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
     6361 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     6421 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     6481 cgggtttgcc gccagaacac aggatcctcg agccaccatg gccaagcctt tgtctcaaga
     6541 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     6601 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     6661 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     6721 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     6781 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     6841 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     6901 tgtgtgggag ggctaagtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     6961 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7021 caaactcatc aatgtatctt atcatgtctg gaattgactc aaatgatgtc aattagtcta
     7081 tcagaagctc atctggtctc ccttccgggg gacaagacat ccctgtttaa tatttaaaca
     7141 gcagtgttcc caaactgggt tcttatatcc cttgctctgg tcaaccaggt tgcagggttt
     7201 cctgtcctca caggaacgaa gtccctaaag aaacagtggc agccaggttt agccccggaa
     7261 ttgactggat tcctttttta gggcccattg gtatggcttt ttccccgtat ccccccaggt
     7321 gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc
     7381 cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc
     7441 ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc
     7501 cctgggggct ttgggggggg gctgtccctg atatctataa caagaaaata tatatataat
     7561 aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta
     7621 aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa
     7681 tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga
     7741 tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga
     7801 atgcatgcgt caattttacg cagactatct ttctagggtt aatctagctg catcaggatc
     7861 atatcgtcgg gtcttttttc cggctcagtc atcgcccaag ctggcgctat ctgggcatcg
     7921 gggaggaaga agcccgtgcc ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg
     7981 aggatgactg ctgctgcatt gacgttgagc gaaaacgcac gtttaccatg atgattcggg
     8041 aaggtgtggc catgcacgcc tttaacggtg aactgttcgt tcaggccacc tgggatacca
     8101 gttcgtcgcg gcttttccgg acacagttcc ggatggtcag cccgaagcgc atcagcaacc
     8161 cgaacaatac cggcgacagc cggaactgcc gtgccggtgt gcagattaat gacagcggtg
     8221 cggcgctggg atattacgtc agcgaggacg ggtatcctgg ctggatgccg cagaaatgga
     8281 catggatacc ccgtgagtta cccggcgggc gcgcttggcg taatcatggt catagctgtt
     8341 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     8401 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     8461 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     8521 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     8581 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     8641 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     8701 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     8761 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     8821 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     8881 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     8941 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     9001 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     9061 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     9121 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     9181 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     9241 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     9301 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     9361 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     9421 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     9481 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     9541 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     9601 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     9661 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     9721 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     9781 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     9841 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     9901 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     9961 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    10021 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
    10081 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    10141 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    10201 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    10261 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    10321 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    10381 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    10441 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
    10501 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
    10561 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
    10621 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
    10681 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
    10741 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
    10801 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
    10861 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
    10921 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
    10981 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
    11041 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
    11101 gcctcgttca ttcacgtttt tgaacccgtg gaggacgggc agactcgcgg tgcaaatgtg
    11161 ttttacagcg tgatggagca gatgaagatg ctcgacacgc tgcagaacac gcagctagat
    11221 taaccctaga aagataatca tattgtgacg tacgttaaag ataatcatgt gtaaaattga
    11281 cgcatgtgtt ttatcggtct gtatatcgag gtttatttat taatttgaat agatattaag
    11341 ttttattata tttacactta catactaata ataaattcaa caaacaattt atttatgttt
    11401 atttatttat taaaaaaaac aaaaactcaa aatttcttct ataaagtaac aaaactttta
    11461 tgagggacag ccccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg
    11521 ggcagcagcg agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc
    11581 cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga
    11641 acgcttctcg ctgctctttg agcctgcaga cacctggggg gatacgggga aaaggcctcc
    11701 acggcc
//
