LOCUS       sequence-314831-       11986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobile CRISPRi sfGFP 'test', gmc6 sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11986)
  AUTHORS   Banta AB, Ward RD, Tran JS, Bacon EE, Peters JM
  TITLE     Programmable Gene Knockdown in Diverse Bacteria Using 
            Mobile-CRISPRi.
  JOURNAL   Curr Protoc Microbiol. 2020 Dec;59(1):e130. doi: 10.1002/cpmc.130.
  PUBMED    33332762
REFERENCE   2  (bases 1 to 11986)
  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: "Curr Protoc
            Microbiol."; date: "2020-12"; volume: "59(1):e130. doi"; pages: " 
            10.1002/cpmc.130"
COMMENT     Sequence Label: pJMP2834
FEATURES             Location/Qualifiers
     source          1..11986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..78
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             79..1161
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1446..1462
                     /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)."
     CDS             join(1818..1820,1821..2534)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1818..2531
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1848..1876)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2471..2491
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(2676..2723)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        join(2800..2805,2806..2822,2823..2828)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2939..3733
                     /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(2993..3012)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3603..3622
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(3785..3832)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     terminator      complement(3883..3969)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(4072..4166)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4194..4210)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4195..4211)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4625..4644)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4744..4766
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4804..4822)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4890..4994
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4995..5063,5064..5855)
                     /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     complement(5213..5232)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6156..6544
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6813..6921
                     /note="incP origin of transfer"
     mobile_element  7025..7190
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(7345..7434)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(7444..9027,9028..9030,9031..11517,
                     11518..11520,11521..11547))
                     /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"
     protein_bind    11589..11605
                     /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)."
ORIGIN
        1 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gatggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag atagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     1201 accccacccg ggtccttgag ccccattatc acctccaaag ccttctcgtc tggtcagttt
     1261 cacctgtttt acgtaaaaac ccgcttcggc gggtttttac ttttggggaa acacagaaaa
     1321 aagcccgcac ctgacagtgc gggctttttt tttcgaccaa aggtgcgact actcttgcct
     1381 actacctatc gactgagctg aaagaattcc taaagatcta taaatgtgag cggataacat
     1441 tgacattgtg agcggataac aagatacttc tagtcatcta attcaacaag aattgtttaa
     1501 gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat caacttgaaa
     1561 aagtggcacc gagtcggtgc tttttttttg aattcatgtg gctgaccgtt ctgttgtctc
     1621 tcgctcttcc gagtagacga acaataaggc ctccctaacg gggggccttt tttattgata
     1681 acaaaagtca gtgcttccgc tatttccaaa ataccgggct aatacggttt aaacgaaaat
     1741 ttatcaaaaa gagtattgac ttaaagtcta acctatagga tacttacagc cagatctgag
     1801 cgagaaggag gtaaagtatg agcaaaggag aagaactttt cactggagtt gtcccaattc
     1861 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtccgtgga gagggtgaag
     1921 gtgatgctac aaacggaaaa ctcaccctta aatttatttg cactactgga aaactacctg
     1981 ttccgtggcc aacacttgtc actactctga cctatggtgt tcaatgcttt tcccgttatc
     2041 cggatcacat gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg
     2101 aacgcactat atctttcaaa gatgacggga cctacaagac gcgtgctgaa gtcaagtttg
     2161 aaggtgatac ccttgttaat cgtatcgagt taaagggtat tgattttaaa gaagatggaa
     2221 acattcttgg acacaaactg gagtacaact ttaactcaca caatgtatac atcacggcag
     2281 acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacgtt gaagatggtt
     2341 ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     2401 taccagacaa ccattacctg tcgacacaat ctgtcctttc gaaagatccc aacgaaaagc
     2461 gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg
     2521 agctctacaa ataataagtt taaacgacct cctggatttg ctcagacagc cttttcgtca
     2581 ttcgtttcag ccaaaaaact taagaccgcc ggtcttgtcc actaccttgc agtaatgcgg
     2641 tggacaggat cggcggtttt cttttctctt ctcaagaagt tcctatactt tctagagaat
     2701 aggaacttcg gaataggaac ttcctcctga acggccataa gaacgaaggc tgtctgttga
     2761 actctcgagc cgtggaaacg gatgaaggca cgaacccagt tgacataagc ctgttcggtt
     2821 cgtaaactgt aatgcaagta gcgtatgcgc tcaaaactgg atggctttct tgccgccaag
     2881 gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgaggat cgtttcgcat
     2941 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3001 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3061 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3121 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3181 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3241 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3301 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3361 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3421 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     3481 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3541 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3601 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3661 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3721 cgagttcttc tgagcgggac tctggggttc gaaatgactc gagcatcagc ttcaaaagcg
     3781 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggtact tcaagatccc
     3841 caattcgagc tcatcgtccg ggccgcaagc tcctagcggc ggatttgtcc tactcaggag
     3901 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
     3961 ttttatttga tgcctcaagc tagagagtca ttaccccagg cgtttaaggg caccaataac
     4021 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtctagcttg gattctcacc
     4081 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     4141 cattactgga tctatcaaca ggagtccaag ctcagctaat taagctagct tatcgatacc
     4201 gtcgacctcg aaccccacgc ccctctttaa tacgacgggc aatttgcact tcagaaaatg
     4261 aagagtttgc tttagccata acaaaagtcc agtatgcttt ttcacagcat aactggactg
     4321 atttcagttt acaactattc tgtctagttt aagactttat tgtcatagtt tagatctatt
     4381 ttgttcagtt taagacttta ttgtccgccc acacctgctt acgcagggca tccatttatt
     4441 actcaaccgt aaccgatttt gccaggttac gcggctggtc ctctagctgg cgtaatagcg
     4501 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     4561 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     4621 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4681 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4741 tctccgggag ctgcatgtgt cagaggtttt caccgtcgtc accgaaacgc gcgagacgaa
     4801 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4861 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4921 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4981 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5041 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5101 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5161 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5221 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5281 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5341 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5401 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5461 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5521 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5581 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5641 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5701 gtgagcgtgg gtcccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5761 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5821 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5881 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5941 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6001 ccgtagaaaa gatcaaaggc agttattggt gcctcactga ttaagcattg gtaactgtca
     6061 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttttgcgg
     6121 ccgcaagatc cggccacgat gcgtccggcg tagaggatct gaagatcagc agttcaacct
     6181 gttgatagta cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact
     6241 aagctctcat gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa
     6301 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat
     6361 aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat
     6421 aaggctttta aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca
     6481 ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct
     6541 gacatgggaa ttagccatgg catcacagta tcgtgatgac agaggcaggg agtgcggcct
     6601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatcag
     6661 gggattcctt aaggtatact ttccgctgca taaccctgct tcggggtcat tatagcgatt
     6721 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     6781 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     6841 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     6901 cctgctctgc gaggctggcc gataagctct gataccgctc gccgcagccg aacgaccgag
     6961 cgcagcgggt cagtgagcga ggaagcggaa gagcccaaac ataacaggaa gaaaaatgcc
     7021 ccgctgtggg cggacaaaat agttgggaac tgggaggggt ggaaatggag tttttaagga
     7081 ttatttaggg aagagtgaca aaatagatgg gaactgggtg tagcgtcgta agctaatacg
     7141 aaaattaaaa atgacaaaat agtttggaac tagatttcac ttatctggtt cttgaggcgg
     7201 ttaaaagagc cgtactcttc tccgatgtcg actaggccat gatgctcatt ctgtgggacc
     7261 aaaacgaaaa aacacccttt cgggtgtctt ttctggaatt tggtaccgag gctgcaagtc
     7321 ccattaagga ggcgcgcctt attacagatc ttcctcagag atgagcttct gttccagatc
     7381 ttcttctgaa atcaactttt gttccagatc ttcttcagag atgagtttct gctccgcggc
     7441 cgcgtcacct cctagctgac tcaaatcaat gcgtgtttca taaagaccag tgatggattg
     7501 atggataaga gtggcatcta aaacttcttt tgtagacgta tatcgtttac gatcaattgt
     7561 tgtatcaaaa tatttaaaag cagcgggagc tccaagattc gtcaacgtaa ataaatgaat
     7621 aatattttct gcttgttcac gtattggttt gtctctatgt ttgttatatg cactaagaac
     7681 tttatctaaa ttggcatctg ctaaaataac acgcttagaa aattcactga tttgctcaat
     7741 aatctcatct aaataatgct tatgctgctc cacaaacaat tgtttttgtt cgttatcttc
     7801 tggactaccc ttcaactttt cataatgact agctaaatat aaaaaattca catatttgct
     7861 tggcagagcc agctcatttc ctttttgtaa ttctccggca ctagccagca tccgtttacg
     7921 accgttttct aactcaaaaa gactatattt aggtagttta atgattaagt cttttttaac
     7981 ttccttatat cctttagctt ctaaaaagtc aatcggattt ttttcaaagg aacttctttc
     8041 cataattgtg atccctagta actctttaac ggattttaac ttcttcgatt tccctttttc
     8101 caccttagca accactagga ctgaataagc taccgttgga ctatcaaaac caccatattt
     8161 ttttggatcc cagtcttttt tacgagcaat aagcttgtcc gaatttcttt ttggtaaaat
     8221 tgactccttg gagaatccgc ctgtctgtac ttctgttttc ttgacaatat tgacttgggg
     8281 catggacaat actttgcgca ctgtggcaaa atctcgccct ttatcccaga caatttctcc
     8341 agtttcccca ttagtttcga ttagagggcg tttgcgaatc tctccatttg caagtgtaat
     8401 ttctgttttg aagaagttca tgatattaga gtaaaagaaa tattttgcgg ttgctttgcc
     8461 tatttcttgc tcagacttag caatcatttt acgaacatca taaactttat aatcaccata
     8521 gacaaactcc gattcaagtt ttggatattt cttaatcaaa gcagttccaa cgacggcatt
     8581 tagatacgca tcatgggcat gatggtaatt gttaatctca cgtactttat agaattggaa
     8641 atcttttcgg aagtcagaaa ctaatttaga ttttaaggta atcactttaa cctctcgaat
     8701 aagtttatca ttttcatcgt atttagtatt catgcgacta tccaaaattt gtgccacatg
     8761 cttagtgatt tggcgagttt caaccaattg gcgtttgata aaaccagctt tatcaagttc
     8821 actcaaacct ccacgttcag ctttcgttaa attatcaaac ttacgttgag tgattaactt
     8881 ggcgtttaga agttgtctcc aatagttttt catctttttg actacttctt cacttggaac
     8941 gttatccgat ttaccacgat ttttatcaga acgcgttaag accttattgt ctattgaatc
     9001 gtctttaagg aaactttgtg gaacaatggc atcgacatca taatcactta aacgattaat
     9061 atctaattct tggtccacat acatgtctct tccattttgg agataataga gatagagctt
     9121 ttcattttgc aattgagtat tttcaacagg atgctcttta agaatctgac ttcctaattc
     9181 tttgatacct tcttcgattc gtttcatacg ctctcgcgaa tttttctggc ccttttgagt
     9241 tgtctgattt tcacgtgcca tttcaataac gatattttct ggcttatgcc gccccattac
     9301 tttgaccaat tcatcaacaa cttttacagt ctgtaaaata ccttttttaa tagcagggct
     9361 accagctaaa tttgcaatat gttcatgtaa actatcgcct tgtccagaca cttgtgcttt
     9421 ttgaatatct tctttaaatg tcaaactatc atcatggatc agctgcataa aattgcgatt
     9481 ggcaaaacca tctgatttca aaaaatctaa tattgttttg ccagattgct tatccctaat
     9541 accattaatc aattttcgag acaaacgtcc ccaaccagta taacggcgac gtttaagctg
     9601 tttcatcacc ttatcatcaa agaggtgagc atatgtttta agtctttcct caatcatctc
     9661 cctatcttca aataaggtca atgttaaaac aatatcctct aagatatctt cattttcttc
     9721 attatccaaa aaatctttat ctttaataat ttttagcaaa tcatggtagg tacctaatga
     9781 agcattaaat ctatcttcaa ctcctgaaat ttcaacacta tcaaaacatt ctattttttt
     9841 gaaataatct tcttttaatt gcttaacggt tacttttcga tttgttttga agagtaaatc
     9901 aacaatggct ttcttctgtt cacctgaaag aaatgctggt tttcgcattc cttcagtaac
     9961 atatttgacc tttgtcaatt cgttataaac cgtaaaatac tcataaagca aactatgttt
    10021 tggtagtact ttttcatttg gaagattttt atcaaagttt gtcatgcgtt caataaatga
    10081 ttgagctgaa gcacctttat cgacaacttc ttcaaaattc catggggtaa ttgtttcttc
    10141 agacttccga gtcatccatg caaaacgact attgccacgc gccaatggac caacataata
    10201 agggattcga aaagtcaaga ttttttcaat cttctcacga ttgtctttta aaaatggata
    10261 aaaatcttct tgtcgtctca aaatagcatg cagctcaccc aagtgaattt gatggggaat
    10321 agagccgttg tcaaaggtcc gttgcttgcg cagcaaatct tcacgattta gtttcaccaa
    10381 taattcctca gtaccatcca ttttttctaa aattggtttg ataaatttat aaaattcttc
    10441 ttggctagct cccccatcaa tataacctgc atatccgttt tttgattgat caaaaaagat
    10501 ttctttatac ttttctggaa gttgttgtcg aactaaagct tttaaaagag tcaagtcttg
    10561 atgatgttca tcgtagcgtt taatcattga agctgatagg ggagccttag ttatttcagt
    10621 atttactctt aggatatctg aaagtaaaat agcatctgat aaattcttag ctgccaaaaa
    10681 caaatcagca tattgatctc caatttgcgc caataaatta tctaaatcat catcgtaagt
    10741 atcttttgaa agctgtaatt tagcatcttc tgccaaatca aaatttgatt taaaattagg
    10801 ggtcaaaccc aatgacaaag caatgagatt cccaaataag ccatttttct tctcaccggg
    10861 gagctgagca atgagatttt ctaatcgtct tgatttactc aatcgtgcag aaagaatcgc
    10921 tttagcatct actccacttg cgttaatagg gttttcttca aataattgat tgtaggtttg
    10981 taccaactgg ataaatagtt tgtccacatc actattatca ggatttaaat ctccctcaat
    11041 caaaaaatga ccacgaaact taatcatatg cgctaaggcc aaatagatta agcgcaaatc
    11101 cgctttatca gtagaatcta ccaatttttt tcgcagatga tagatagttg gatatttctc
    11161 atgataagca acttcatcta ctatatttcc aaaaatagga tgacgttcat gcttcttatc
    11221 ttcttccacc aaaaaagact cttcaagtcg atgaaagaaa ctatcatcta ctttcgccat
    11281 ctcatttgaa aaaatctcct gtagataaca aatacgattc ttccgacgtg tataccttct
    11341 acgagctgtc cgtttgagac gagtcgcttc cgctgtctct ccactgtcaa ataaaagagc
    11401 ccctataaga ttttttttga tactgtggcg gtctgtattt cccagaacct tgaacttttt
    11461 agacggaacc ttatattcat cagtgatcac cgcccatccg acgctatttg tgccgatagc
    11521 taagcctatt gagtatttct tatccatact agttttcctg tgtgaaacct gctgatgtgc
    11581 tcagtatctt gttatccgct cacaatgtca atgttatccg ctcacattta taatatttta
    11641 tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
    11701 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
    11761 tgaaccgagg taactggctt ggtgataagc tgtcaaacca gatcaattcg cactagtctg
    11821 cagacgtaaa aaaagcggcg tggttagccg cttttttaat tgccggagcg caataaaaaa
    11881 gcccccggaa ggtgatcttc cgggggcttt ctcatgcgtt gcttgtattg aaaatggggg
    11941 ccatgcctac ctatctgcct cccgggaagc ggcatgcatt tacgtt
//
