LOCUS       sequence-97051-e       10113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-CibN in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10113)
  AUTHORS   Polstein LR, Gersbach CA
  TITLE     A light-inducible CRISPR-Cas9 system for control of endogenous gene 
            activation.
  JOURNAL   Nat Chem Biol. 2015 Feb 9. doi: 10.1038/nchembio.1753.
  PUBMED    25664691
REFERENCE   2  (bases 1 to 10113)
  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: "Nat Chem 
            Biol. 2015 Feb 9. doi: 10.1038/nchembio.1753."
COMMENT     Sequence Label: pcDNA3.1-dCas9-CibN
FEATURES             Location/Qualifiers
     source          1..10113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        250..453
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     404..424
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     498..517
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        498..516
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             542..607
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             614..634
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(644..670,671..673,674..3160,3161..3163,3164..4747)
                     /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             4772..4792
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4805..5314
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKKRKFDTETKDCN
                     EKKKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     primer_bind     complement(5342..5359)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5348..5572
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5618..6046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5705..5724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5915..5936
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6055..6075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6060..6389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6240..6375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6302..6321
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6456..7250
                     /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(6510..6529)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7120..7139
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7424..7545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7461..7480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7515..7534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7594..7610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7594..7610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7607..7629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7618..7634
                     /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(7642..7648,7649..7666,7667..7672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7687..7708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7825..7842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7996..8581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8076..8095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8752..9543,9544..9612))
                     /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     9375..9394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9613..9717)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9792..9811)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(9983..10113,1..249)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
       61 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      121 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      181 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      241 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      301 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      361 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      421 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      481 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgccat
      541 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga
      601 tgacaagatg gcccccaaga agaagaggaa ggtgggccgc ggaatggata agaaatactc
      661 aataggctta gctatcggca caaatagcgt cggatgggcg gtgatcactg atgaatataa
      721 ggttccgtct aaaaagttca aggttctggg aaatacagac cgccacagta tcaaaaaaaa
      781 tcttataggg gctcttttat ttgacagtgg agagacagcg gaagcgactc gtctcaaacg
      841 gacagctcgt agaaggtata cacgtcggaa gaatcgtatt tgttatctac aggagatttt
      901 ttcaaatgag atggcgaaag tagatgatag tttctttcat cgacttgaag agtctttttt
      961 ggtggaagaa gacaagaagc atgaacgtca tcctattttt ggaaatatag tagatgaagt
     1021 tgcttatcat gagaaatatc caactatcta tcatctgcga aaaaaattgg tagattctac
     1081 tgataaagcg gatttgcgct taatctattt ggccttagcg catatgatta agtttcgtgg
     1141 tcattttttg attgagggag atttaaatcc tgataatagt gatgtggaca aactatttat
     1201 ccagttggta caaacctaca atcaattatt tgaagaaaac cctattaacg caagtggagt
     1261 agatgctaaa gcgattcttt ctgcacgatt gagtaaatca agacgattag aaaatctcat
     1321 tgctcagctc cccggtgaga agaaaaatgg cttatttggg aatctcattg ctttgtcatt
     1381 gggtttgacc cctaatttta aatcaaattt tgatttggca gaagatgcta aattacagct
     1441 ttcaaaagat acttacgatg atgatttaga taatttattg gcgcaaattg gagatcaata
     1501 tgctgatttg tttttggcag ctaagaattt atcagatgct attttacttt cagatatcct
     1561 aagagtaaat actgaaataa ctaaggctcc cctatcagct tcaatgatta aacgctacga
     1621 tgaacatcat caagacttga ctcttttaaa agctttagtt cgacaacaac ttccagaaaa
     1681 gtataaagaa atcttttttg atcaatcaaa aaacggatat gcaggttata ttgatggggg
     1741 agctagccaa gaagaatttt ataaatttat caaaccaatt ttagaaaaaa tggatggtac
     1801 tgaggaatta ttggtgaaac taaatcgtga agatttgctg cgcaagcaac ggacctttga
     1861 caacggctct attccccatc aaattcactt gggtgagctg catgctattt tgagaagaca
     1921 agaagacttt tatccatttt taaaagacaa tcgtgagaag attgaaaaaa tcttgacttt
     1981 tcgaattcct tattatgttg gtccattggc gcgtggcaat agtcgttttg catggatgac
     2041 tcggaagtct gaagaaacaa ttaccccatg gaattttgaa gaagttgtcg ataaaggtgc
     2101 ttcagctcaa tcatttattg aacgcatgac aaactttgat aaaaatcttc caaatgaaaa
     2161 agtactacca aaacatagtt tgctttatga gtattttacg gtttataacg aattgacaaa
     2221 ggtcaaatat gttactgaag gaatgcgaaa accagcattt ctttcaggtg aacagaagaa
     2281 agccattgtt gatttactct tcaaaacaaa tcgaaaagta accgttaagc aattaaaaga
     2341 agattatttc aaaaaaatag aatgttttga tagtgttgaa atttcaggag ttgaagatag
     2401 atttaatgct tcattaggta cctaccatga tttgctaaaa attattaaag ataaagattt
     2461 tttggataat gaagaaaatg aagatatctt agaggatatt gttttaacat tgaccttatt
     2521 tgaagatagg gagatgattg aggaaagact taaaacatat gctcacctct ttgatgataa
     2581 ggtgatgaaa cagcttaaac gtcgccgtta tactggttgg ggacgtttgt ctcgaaaatt
     2641 gattaatggt attagggata agcaatctgg caaaacaata ttagattttt tgaaatcaga
     2701 tggttttgcc aatcgcaatt ttatgcagct gatccatgat gatagtttga catttaaaga
     2761 agacattcaa aaagcacaag tgtctggaca aggcgatagt ttacatgaac atattgcaaa
     2821 tttagctggt agccctgcta ttaaaaaagg tattttacag actgtaaaag ttgttgatga
     2881 attggtcaaa gtaatggggc ggcataagcc agaaaatatc gttattgaaa tggcacgtga
     2941 aaatcagaca actcaaaagg gccagaaaaa ttcgcgagag cgtatgaaac gaatcgaaga
     3001 aggtatcaaa gaattaggaa gtcagattct taaagagcat cctgttgaaa atactcaatt
     3061 gcaaaatgaa aagctctatc tctattatct ccaaaatgga agagacatgt atgtggacca
     3121 agaattagat attaatcgtt taagtgatta tgatgtcgat gccattgttc cacaaagttt
     3181 ccttaaagac gattcaatag acaataaggt cttaacgcgt tctgataaaa atcgtggtaa
     3241 atcggataac gttccaagtg aagaagtagt caaaaagatg aaaaactatt ggagacaact
     3301 tctaaacgcc aagttaatca ctcaacgtaa gtttgataat ttaacgaaag ctgaacgtgg
     3361 aggtttgagt gaacttgata aagctggttt tatcaaacgc caattggttg aaactcgcca
     3421 aatcactaag catgtggcac aaattttgga tagtcgcatg aatactaaat acgatgaaaa
     3481 tgataaactt attcgagagg ttaaagtgat taccttaaaa tctaaattag tttctgactt
     3541 ccgaaaagat ttccaattct ataaagtacg tgagattaac aattaccatc atgcccatga
     3601 tgcgtatcta aatgccgtcg ttggaactgc tttgattaag aaatatccaa aacttgaatc
     3661 ggagtttgtc tatggtgatt ataaagttta tgatgttcgt aaaatgattg ctaagtctga
     3721 gcaagaaata ggcaaagcaa ccgcaaaata tttcttttac tctaatatca tgaacttctt
     3781 caaaacagaa attacacttg caaatggaga gattcgcaaa cgccctctaa tcgaaactaa
     3841 tggggaaact ggagaaattg tctgggataa agggcgagat tttgccacag tgcgcaaagt
     3901 attgtccatg ccccaagtca atattgtcaa gaaaacagaa gtacagacag gcggattctc
     3961 caaggagtca attttaccaa aaagaaattc ggacaagctt attgctcgta aaaaagactg
     4021 ggatccaaaa aaatatggtg gttttgatag tccaacggta gcttattcag tcctagtggt
     4081 tgctaaggtg gaaaaaggga aatcgaagaa gttaaaatcc gttaaagagt tactagggat
     4141 cacaattatg gaaagaagtt cctttgaaaa aaatccgatt gactttttag aagctaaagg
     4201 atataaggaa gttaaaaaag acttaatcat taaactacct aaatatagtc tttttgagtt
     4261 agaaaacggt cgtaaacgga tgctggctag tgccggagaa ttacaaaaag gaaatgagct
     4321 ggctctgcca agcaaatatg tgaatttttt atatttagct agtcattatg aaaagttgaa
     4381 gggtagtcca gaagataacg aacaaaaaca attgtttgtg gagcagcata agcattattt
     4441 agatgagatt attgagcaaa tcagtgaatt ttctaagcgt gttattttag cagatgccaa
     4501 tttagataaa gttcttagtg catataacaa acatagagac aaaccaatac gtgaacaagc
     4561 agaaaatatt attcatttat ttacgttgac gaatcttgga gctcccgctg cttttaaata
     4621 ttttgataca acaattgatc gtaaacgata tacgtctaca aaagaagttt tagatgccac
     4681 tcttatccat caatccatca ctggtcttta tgaaacacgc attgatttga gtcagctagg
     4741 aggtgaccca attgccggat ccaaggctag cccgaaaaag aaacgcaaag ttgggcgcgc
     4801 caccatgaat ggagctatag gaggtgacct tttgctcaat tttcctgaca tgtcggtcct
     4861 agagcgccaa agggctcacc tcaagtacct caatcccacc tttgattctc ctctcgccgg
     4921 cttctttgcc gattcttcaa tgattaccgg cggcgagatg gacagctatc tttcgactgc
     4981 cggtttgaat cttccgatga tgtacggtga gacgacggtg gaaggtgatt caagactctc
     5041 aatttcgccg gaaacgacgc ttgggactgg aaatttcaag aaacggaagt ttgatacaga
     5101 gactaaggat tgtaatgaga agaagaagaa gatgacgatg aacagagatg acctagtaga
     5161 agaaggagaa gaagagaagt cgaaaataac agagcaaaac aatgggagca caaaaagcat
     5221 caagaagatg aaacacaaag ccaagaaaga agagaacaat ttctctaatg attcatctaa
     5281 agtgacgaag gaattggaga aaacggatta tatttaaact taagtttaaa ccgctgatca
     5341 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     5401 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     5461 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     5521 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     5581 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     5641 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5701 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5761 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5821 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5881 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5941 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     6001 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     6061 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6121 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6181 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6241 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6301 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     6361 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     6421 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     6481 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     6541 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     6601 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     6661 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     6721 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     6781 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6841 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6901 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6961 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     7021 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     7081 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     7141 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     7201 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     7261 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     7321 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     7381 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     7441 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     7501 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     7561 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     7621 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     7681 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     7741 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     7801 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7861 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     7921 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7981 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8041 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8161 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     8221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     8341 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8401 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     8461 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     8521 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8581 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8641 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8701 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8761 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8821 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8881 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8941 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     9001 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     9061 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     9121 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     9181 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     9241 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     9301 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     9361 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     9421 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     9481 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     9541 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     9601 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9661 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9721 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     9781 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     9841 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     9901 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     9961 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
    10021 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
    10081 ggcccgcctg gctgaccgcc caacgacccc cgc
//
