LOCUS       sequence-348230-       10057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VP64 fused to N-terminus of dCas9; pCDNA3 vector backbone, mammalian
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10057)
  AUTHORS   Umeh-Garcia M, O'Geen H, Simion C, Gephart MH, Segal DJ, Sweeney CA
  TITLE     Aberrant promoter methylation contributes to LRIG1 silencing in 
            basal/triple-negative breast cancer.
  JOURNAL   Br J Cancer. 2022 Apr 19. pii: 10.1038/s41416-022-01812-8. doi: 
            10.1038/s41416-022-01812-8.
  PUBMED    35440669
REFERENCE   2  (bases 1 to 10057)
  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: "Br J 
            Cancer. 2022 Apr 19. pii: 10.1038/s41416-022-01812-8. doi: 
            10.1038/s41416-022-01812-8."
COMMENT     Sequence Label: VP64-dCas9
FEATURES             Location/Qualifiers
     source          1..10057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(88..108)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        93..422
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      273..408
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     335..354
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             489..1283
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(543..562)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1153..1172
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1459..1580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1496..1515)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1550..1569
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1629..1645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1629..1645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1642..1664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1653..1669
                     /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(1677..1683,1684..1701,1702..1707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1722..1743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1860..1877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2031..2619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2111..2130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2790..3581,3582..3650))
                     /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     3413..3432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3651..3755)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3830..3849)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4021..4400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4401..4604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4555..4575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4601..4625
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4722..4742
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4761..4826
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(5049..5075,5076..5078,5079..7562,7563..7568,7569..7634,
                     7635..7637,7638..9152)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(5049..5075,5076..5078,5079..7565,7566..7568,
                     7569..9152)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5049..9152
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9228..9275
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9294..9350
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(9414..9433)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    9458..9506
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(9708..10057,1..79)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9795..9814)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10005..10026
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
       61 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
      121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
      181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      241 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
      301 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
      361 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
      421 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
      481 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
      541 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
      601 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
      661 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
      721 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
      781 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
      841 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
      901 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
      961 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     1021 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     1081 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     1141 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     1201 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     1261 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     1321 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     1381 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     1441 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1501 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1561 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     1621 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1681 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1741 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1801 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1861 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1921 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1981 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2041 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2101 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2161 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2221 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2281 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2341 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2401 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2461 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2521 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2581 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2641 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2701 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2761 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2821 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2881 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2941 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3001 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3061 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3121 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3181 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3241 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3301 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3361 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3421 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3481 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3541 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3601 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3661 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3721 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3781 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     3841 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     3901 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     3961 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     4021 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     4081 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4141 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4201 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4261 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4321 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4381 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     4441 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4501 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4561 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
     4621 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
     4681 ccagcctccg gactctagag gatcgaaccc ttgccaccat gcccaagaag aagaggaagg
     4741 tgggaggctc cggaggaagc gactataagg accacgacgg agactacaag gatcatgata
     4801 ttgattacaa agacgatgac gataagggtg gcgggtccgg cggtggatcc ggtaccgacg
     4861 ctttggatga ctttgatttg gacatgctgg atgctctaga tgacttcgac ctggatatgc
     4921 tggacgcact tgacgacttt gacctcgaca tgctagacgc tctggacgac ttcgatctag
     4981 acatgctcag taccggaggg agtggtggaa gcggcggttc tggtggctca ggcggcagcg
     5041 gccggccaat ggacaagaag tactccattg ggctcgctat cggcacaaac agcgtcggct
     5101 gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata
     5161 ccgatcgcca cagcataaag aagaacctca ttggcgccct cctgttcgac tccggggaga
     5221 cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc agaaagaatc
     5281 ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat gactctttct
     5341 tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag cgccacccaa
     5401 tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc atatatcatc
     5461 tgaggaagaa gcttgtagac agtactgata aggctgactt gcggttgatc tatctcgcgc
     5521 tggcgcatat gatcaaattt cggggacact tcctcatcga gggggacctg aacccagaca
     5581 acagcgatgt cgacaaactc tttatccaac tggttcagac ttacaatcag cttttcgaag
     5641 agaacccgat caacgcatcc ggagttgacg ccaaagcaat cctgagcgct aggctgtcca
     5701 aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag aacggcctgt
     5761 ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct aacttcgacc
     5821 tggccgaaga tgccaagctt caactgagca aagacaccta cgatgatgat ctcgacaatc
     5881 tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag aacctgtcag
     5941 acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa gctccgctga
     6001 gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg ctgaaggccc
     6061 ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag tctaaaaatg
     6121 gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa tttattaagc
     6181 ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac agagaagatc
     6241 tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt cacctgggcg
     6301 aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa gataacaggg
     6361 aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc ctcgcccggg
     6421 gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact ccctggaact
     6481 tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg atgactaact
     6541 ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact
     6601 tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg agaaagccag
     6661 cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag acgaaccgga
     6721 aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt ttcgactctg
     6781 ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat cacgatctcc
     6841 tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac attcttgagg
     6901 acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa cgcttgaaaa
     6961 cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc cgatatacag
     7021 gatgggggcg gctgtcaaga aaactgatca atgggatccg agacaagcag agtggaaaga
     7081 caatcctgga ttttcttaag tccgatggat ttgccaacag gaacttcatg cagttgatcc
     7141 atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct ggccaggggg
     7201 acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa aagggaatac
     7261 tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat aagcccgaga
     7321 atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag aagaacagta
     7381 gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa atccttaagg
     7441 aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac tacctgcaga
     7501 acggcaggga catgtacgtg gatcaggaac tggacatcaa tcggctctcc gactacgacg
     7561 tggatgctat cgtgccccag tcttttctca aagatgattc tattgataat aaagtgttga
     7621 caagatccga taaaaataga gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga
     7681 aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa cggaagttcg
     7741 ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca
     7801 aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt ctcgattcac
     7861 gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa gttattactc
     7921 tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag gtgagagaga
     7981 tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc actgcactta
     8041 tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg agactataaa gtgtacgatg
     8101 ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct aagtacttct
     8161 tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat ggagagattc
     8221 ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg gacaagggta
     8281 gggatttcgc gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga
     8341 ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg aacagcgaca
     8401 agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc gattctccta
     8461 cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct aaaaaactca
     8521 aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atcaagcttc gaaaaaaacc
     8581 ccatcgactt tctcgaggcg aaaggatata aagaggtcaa aaaagacctc atcattaagc
     8641 ttcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc gctagtgcgg
     8701 gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat ttcttgtatc
     8761 tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag aagcagctgt
     8821 tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc gaattctcca
     8881 aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac aataagcaca
     8941 gggataagcc catcagggag caggcagaaa acattatcca cttgtttact ctgaccaact
     9001 tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag cggtacacct
     9061 ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg ctctatgaaa
     9121 caagaatcga cctctctcag ctcggtggag acggtggtag tggaggttca ggaggatccg
     9181 gggggagcgg agggagcgct agcggtggag gctccggagg tggatctaaa aggccggcgg
     9241 ccacgaaaaa ggccggtcag gcaaaaaaga aaaagggtgg tagtggaagc ggagctacta
     9301 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct gcggccgcat
     9361 gaaagggttc gatccctacc ggttagtaat gagtttaaac gggggaggct aactgaaaca
     9421 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     9481 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     9541 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     9601 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     9661 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     9721 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     9781 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     9841 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     9901 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     9961 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
    10021 aactggaaca acactcaacc ctatctcggt ctattct
//
