LOCUS       sequence-314829-       12118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobile CRISPRi sfGFP 'test', empty sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12118)
  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 12118)
  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: pJMP2824
FEATURES             Location/Qualifiers
     source          1..12118
                     /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(1837..1839,1840..2553)
                     /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             1837..2550
                     /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(1867..1895)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2490..2510
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(2695..2742)
                     /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(2819..2824,2825..2841,2842..2847)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     RBS             2863..2874
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3100..3888
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     protein_bind    complement(3917..3964)
                     /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(4015..4101)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(4204..4298)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4326..4342)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4327..4343)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4757..4776)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4876..4898
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4936..4954)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5022..5126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5127..5195,5196..5987)
                     /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(5345..5364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6288..6676
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6945..7053
                     /note="incP origin of transfer"
     mobile_element  7157..7322
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(7477..7566)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(7576..9159,9160..9162,9163..11649,
                     11650..11652,11653..11679))
                     /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    11721..11737
                     /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 tagttgagac caactttggt ctccaccata
     1501 gcggtcggtc tctgtttaag agctatgctg gaaacagcat agcaagttta aataaggcta
     1561 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttttga attcatgtgg
     1621 ctgaccgttc tgttgtctct cgctcttccg agtagacgaa caataaggcc tccctaacgg
     1681 ggggcctttt ttattgataa caaaagtcag tgcttccgct atttccaaaa taccgggcta
     1741 atacggttta aacgaaaatt tatcaaaaag agtattgact taaagtctaa cctataggat
     1801 acttacagcc agatctgagc gagaaggagg taaagtatga gcaaaggaga agaacttttc
     1861 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
     1921 gtccgtggag agggtgaagg tgatgctaca aacggaaaac tcacccttaa atttatttgc
     1981 actactggaa aactacctgt tccgtggcca acacttgtca ctactctgac ctatggtgtt
     2041 caatgctttt cccgttatcc ggatcacatg aaacggcatg actttttcaa gagtgccatg
     2101 cccgaaggtt atgtacagga acgcactata tctttcaaag atgacgggac ctacaagacg
     2161 cgtgctgaag tcaagtttga aggtgatacc cttgttaatc gtatcgagtt aaagggtatt
     2221 gattttaaag aagatggaaa cattcttgga cacaaactgg agtacaactt taactcacac
     2281 aatgtataca tcacggcaga caaacaaaag aatggaatca aagctaactt caaaattcgc
     2341 cacaacgttg aagatggttc cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     2401 ggcgatggcc ctgtcctttt accagacaac cattacctgt cgacacaatc tgtcctttcg
     2461 aaagatccca acgaaaagcg tgaccacatg gtccttcttg agtttgtaac tgctgctggg
     2521 attacacatg gcatggatga gctctacaaa taataagttt aaacgacctc ctggatttgc
     2581 tcagacagcc ttttcgtcat tcgtttcagc caaaaaactt aagaccgccg gtcttgtcca
     2641 ctaccttgca gtaatgcggt ggacaggatc ggcggttttc ttttctcttc tcaagaagtt
     2701 cctatacttt ctagagaata ggaacttcgg aataggaact tcctcctgaa cggccataag
     2761 aacgaaggct gtctgttgaa ctctcgagcc gtggaaacgg atgaaggcac gaacccagtt
     2821 gacataagcc tgttcggttc gtaaactgta atgcagtgat ctaaagagga gaaaggatct
     2881 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     2941 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     3001 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     3061 acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc
     3121 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     3181 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     3241 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     3301 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     3361 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     3421 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     3481 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     3541 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     3601 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     3661 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     3721 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     3781 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     3841 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaatc tcgagcatca
     3901 gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggta
     3961 cttcaagatc cccaattcga gctcatcgtc cgggccgcaa gctcctagcg gcggatttgt
     4021 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     4081 ctgagccttt cgttttattt gatgcctcaa gctagagagt cattacccca ggcgtttaag
     4141 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtctagct
     4201 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     4261 gagttctgag gtcattactg gatctatcaa caggagtcca agctcagcta attaagctag
     4321 cttatcgata ccgtcgacct cgaaccccac gcccctcttt aatacgacgg gcaatttgca
     4381 cttcagaaaa tgaagagttt gctttagcca taacaaaagt ccagtatgct ttttcacagc
     4441 ataactggac tgatttcagt ttacaactat tctgtctagt ttaagacttt attgtcatag
     4501 tttagatcta ttttgttcag tttaagactt tattgtccgc ccacacctgc ttacgcaggg
     4561 catccattta ttactcaacc gtaaccgatt ttgccaggtt acgcggctgg tcctctagct
     4621 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4681 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4741 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     4801 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4861 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtcg tcaccgaaac
     4921 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4981 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5041 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5101 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5161 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5221 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5281 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5341 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5401 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5461 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5521 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5581 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5641 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5701 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5761 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5821 aatctggagc cggtgagcgt gggtcccgcg gtatcattgc agcactgggg ccagatggta
     5881 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5941 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6001 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6061 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6121 gagcgtcaga ccccgtagaa aagatcaaag gcagttattg gtgcctcact gattaagcat
     6181 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6241 taatttttgc ggccgcaaga tccggccacg atgcgtccgg cgtagaggat ctgaagatca
     6301 gcagttcaac ctgttgatag tacgtactaa gctctcatgt ttcacgtact aagctctcat
     6361 gtttaacgta ctaagctctc atgtttaacg aactaaaccc tcatggctaa cgtactaagc
     6421 tctcatggct aacgtactaa gctctcatgt ttcacgtact aagctctcat gtttgaacaa
     6481 taaaattaat ataaatcagc aacttaaata gcctctaagg ttttaagttt tataagaaaa
     6541 aaaagaatat ataaggcttt taaagctttt aaggtttaac ggttgtggac aacaagccag
     6601 ggatgtaacg cactgagaag cccttagagc ctctcaaagc aattttgagt gacacaggaa
     6661 cacttaacgg ctgacatggg aattagccat ggcatcacag tatcgtgatg acagaggcag
     6721 ggagtgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6781 ctgcgttatc aggggattcc ttaaggtata ctttccgctg cataaccctg cttcggggtc
     6841 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     6901 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     6961 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
     7021 ctcaacggga atcctgctct gcgaggctgg ccgataagct ctgataccgc tcgccgcagc
     7081 cgaacgaccg agcgcagcgg gtcagtgagc gaggaagcgg aagagcccaa acataacagg
     7141 aagaaaaatg ccccgctgtg ggcggacaaa atagttggga actgggaggg gtggaaatgg
     7201 agtttttaag gattatttag ggaagagtga caaaatagat gggaactggg tgtagcgtcg
     7261 taagctaata cgaaaattaa aaatgacaaa atagtttgga actagatttc acttatctgg
     7321 ttcttgaggc ggttaaaaga gccgtactct tctccgatgt cgactaggcc atgatgctca
     7381 ttctgtggga ccaaaacgaa aaaacaccct ttcgggtgtc ttttctggaa tttggtaccg
     7441 aggctgcaag tcccattaag gaggcgcgcc ttattacaga tcttcctcag agatgagctt
     7501 ctgttccaga tcttcttctg aaatcaactt ttgttccaga tcttcttcag agatgagttt
     7561 ctgctccgcg gccgcgtcac ctcctagctg actcaaatca atgcgtgttt cataaagacc
     7621 agtgatggat tgatggataa gagtggcatc taaaacttct tttgtagacg tatatcgttt
     7681 acgatcaatt gttgtatcaa aatatttaaa agcagcggga gctccaagat tcgtcaacgt
     7741 aaataaatga ataatatttt ctgcttgttc acgtattggt ttgtctctat gtttgttata
     7801 tgcactaaga actttatcta aattggcatc tgctaaaata acacgcttag aaaattcact
     7861 gatttgctca ataatctcat ctaaataatg cttatgctgc tccacaaaca attgtttttg
     7921 ttcgttatct tctggactac ccttcaactt ttcataatga ctagctaaat ataaaaaatt
     7981 cacatatttg cttggcagag ccagctcatt tcctttttgt aattctccgg cactagccag
     8041 catccgttta cgaccgtttt ctaactcaaa aagactatat ttaggtagtt taatgattaa
     8101 gtctttttta acttccttat atcctttagc ttctaaaaag tcaatcggat ttttttcaaa
     8161 ggaacttctt tccataattg tgatccctag taactcttta acggatttta acttcttcga
     8221 tttccctttt tccaccttag caaccactag gactgaataa gctaccgttg gactatcaaa
     8281 accaccatat ttttttggat cccagtcttt tttacgagca ataagcttgt ccgaatttct
     8341 ttttggtaaa attgactcct tggagaatcc gcctgtctgt acttctgttt tcttgacaat
     8401 attgacttgg ggcatggaca atactttgcg cactgtggca aaatctcgcc ctttatccca
     8461 gacaatttct ccagtttccc cattagtttc gattagaggg cgtttgcgaa tctctccatt
     8521 tgcaagtgta atttctgttt tgaagaagtt catgatatta gagtaaaaga aatattttgc
     8581 ggttgctttg cctatttctt gctcagactt agcaatcatt ttacgaacat cataaacttt
     8641 ataatcacca tagacaaact ccgattcaag ttttggatat ttcttaatca aagcagttcc
     8701 aacgacggca tttagatacg catcatgggc atgatggtaa ttgttaatct cacgtacttt
     8761 atagaattgg aaatcttttc ggaagtcaga aactaattta gattttaagg taatcacttt
     8821 aacctctcga ataagtttat cattttcatc gtatttagta ttcatgcgac tatccaaaat
     8881 ttgtgccaca tgcttagtga tttggcgagt ttcaaccaat tggcgtttga taaaaccagc
     8941 tttatcaagt tcactcaaac ctccacgttc agctttcgtt aaattatcaa acttacgttg
     9001 agtgattaac ttggcgttta gaagttgtct ccaatagttt ttcatctttt tgactacttc
     9061 ttcacttgga acgttatccg atttaccacg atttttatca gaacgcgtta agaccttatt
     9121 gtctattgaa tcgtctttaa ggaaactttg tggaacaatg gcatcgacat cataatcact
     9181 taaacgatta atatctaatt cttggtccac atacatgtct cttccatttt ggagataata
     9241 gagatagagc ttttcatttt gcaattgagt attttcaaca ggatgctctt taagaatctg
     9301 acttcctaat tctttgatac cttcttcgat tcgtttcata cgctctcgcg aatttttctg
     9361 gcccttttga gttgtctgat tttcacgtgc catttcaata acgatatttt ctggcttatg
     9421 ccgccccatt actttgacca attcatcaac aacttttaca gtctgtaaaa tacctttttt
     9481 aatagcaggg ctaccagcta aatttgcaat atgttcatgt aaactatcgc cttgtccaga
     9541 cacttgtgct ttttgaatat cttctttaaa tgtcaaacta tcatcatgga tcagctgcat
     9601 aaaattgcga ttggcaaaac catctgattt caaaaaatct aatattgttt tgccagattg
     9661 cttatcccta ataccattaa tcaattttcg agacaaacgt ccccaaccag tataacggcg
     9721 acgtttaagc tgtttcatca ccttatcatc aaagaggtga gcatatgttt taagtctttc
     9781 ctcaatcatc tccctatctt caaataaggt caatgttaaa acaatatcct ctaagatatc
     9841 ttcattttct tcattatcca aaaaatcttt atctttaata atttttagca aatcatggta
     9901 ggtacctaat gaagcattaa atctatcttc aactcctgaa atttcaacac tatcaaaaca
     9961 ttctattttt ttgaaataat cttcttttaa ttgcttaacg gttacttttc gatttgtttt
    10021 gaagagtaaa tcaacaatgg ctttcttctg ttcacctgaa agaaatgctg gttttcgcat
    10081 tccttcagta acatatttga cctttgtcaa ttcgttataa accgtaaaat actcataaag
    10141 caaactatgt tttggtagta ctttttcatt tggaagattt ttatcaaagt ttgtcatgcg
    10201 ttcaataaat gattgagctg aagcaccttt atcgacaact tcttcaaaat tccatggggt
    10261 aattgtttct tcagacttcc gagtcatcca tgcaaaacga ctattgccac gcgccaatgg
    10321 accaacataa taagggattc gaaaagtcaa gattttttca atcttctcac gattgtcttt
    10381 taaaaatgga taaaaatctt cttgtcgtct caaaatagca tgcagctcac ccaagtgaat
    10441 ttgatgggga atagagccgt tgtcaaaggt ccgttgcttg cgcagcaaat cttcacgatt
    10501 tagtttcacc aataattcct cagtaccatc cattttttct aaaattggtt tgataaattt
    10561 ataaaattct tcttggctag ctcccccatc aatataacct gcatatccgt tttttgattg
    10621 atcaaaaaag atttctttat acttttctgg aagttgttgt cgaactaaag cttttaaaag
    10681 agtcaagtct tgatgatgtt catcgtagcg tttaatcatt gaagctgata ggggagcctt
    10741 agttatttca gtatttactc ttaggatatc tgaaagtaaa atagcatctg ataaattctt
    10801 agctgccaaa aacaaatcag catattgatc tccaatttgc gccaataaat tatctaaatc
    10861 atcatcgtaa gtatcttttg aaagctgtaa tttagcatct tctgccaaat caaaatttga
    10921 tttaaaatta ggggtcaaac ccaatgacaa agcaatgaga ttcccaaata agccattttt
    10981 cttctcaccg gggagctgag caatgagatt ttctaatcgt cttgatttac tcaatcgtgc
    11041 agaaagaatc gctttagcat ctactccact tgcgttaata gggttttctt caaataattg
    11101 attgtaggtt tgtaccaact ggataaatag tttgtccaca tcactattat caggatttaa
    11161 atctccctca atcaaaaaat gaccacgaaa cttaatcata tgcgctaagg ccaaatagat
    11221 taagcgcaaa tccgctttat cagtagaatc taccaatttt tttcgcagat gatagatagt
    11281 tggatatttc tcatgataag caacttcatc tactatattt ccaaaaatag gatgacgttc
    11341 atgcttctta tcttcttcca ccaaaaaaga ctcttcaagt cgatgaaaga aactatcatc
    11401 tactttcgcc atctcatttg aaaaaatctc ctgtagataa caaatacgat tcttccgacg
    11461 tgtatacctt ctacgagctg tccgtttgag acgagtcgct tccgctgtct ctccactgtc
    11521 aaataaaaga gcccctataa gatttttttt gatactgtgg cggtctgtat ttcccagaac
    11581 cttgaacttt ttagacggaa ccttatattc atcagtgatc accgcccatc cgacgctatt
    11641 tgtgccgata gctaagccta ttgagtattt cttatccata ctagttttcc tgtgtgaaac
    11701 ctgctgatgt gctcagtatc ttgttatccg ctcacaatgt caatgttatc cgctcacatt
    11761 tataatattt tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa
    11821 tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag
    11881 ctaccaactc tttgaaccga ggtaactggc ttggtgataa gctgtcaaac cagatcaatt
    11941 cgcactagtc tgcagacgta aaaaaagcgg cgtggttagc cgctttttta attgccggag
    12001 cgcaataaaa aagcccccgg aaggtgatct tccgggggct ttctcatgcg ttgcttgtat
    12061 tgaaaatggg ggccatgcct acctatctgc ctcccgggaa gcggcatgca tttacgtt
//
