LOCUS       sequence-314832-       12099 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 12099)
  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 12099)
  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: pJMP2836
FEATURES             Location/Qualifiers
     source          1..12099
                     /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"
     RBS             2844..2855
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3081..3869
                     /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(3898..3945)
                     /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(3996..4082)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(4185..4279)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4307..4323)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4308..4324)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4738..4757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4857..4879
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4917..4935)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5003..5107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5108..5176,5177..5968)
                     /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(5326..5345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6269..6657
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6926..7034
                     /note="incP origin of transfer"
     mobile_element  7138..7303
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(7458..7547)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(7557..9140,9141..9143,9144..11630,
                     11631..11633,11634..11660))
                     /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    11702..11718
                     /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 aatgcagtga tctaaagagg agaaaggatc tatgcgctca cgcaactggt
     2881 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2941 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     3001 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     3061 ctaaaacaaa gttaaacatc atgagggaag cggtgatcgc cgaagtatcg actcaactat
     3121 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     3181 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     3241 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     3301 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     3361 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     3421 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     3481 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     3541 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     3601 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     3661 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     3721 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     3781 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     3841 aaggcgagat caccaaggta gtcggcaaat ctcgagcatc agcttcaaaa gcgctctgaa
     3901 gttcctatac tttctagaga ataggaactt cggaataggt acttcaagat ccccaattcg
     3961 agctcatcgt ccgggccgca agctcctagc ggcggatttg tcctactcag gagagcgttc
     4021 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     4081 tgatgcctca agctagagag tcattacccc aggcgtttaa gggcaccaat aactgcctta
     4141 aaaaaattac gccccgccct gccactcatc gcagtctagc ttggattctc accaataaaa
     4201 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     4261 ggatctatca acaggagtcc aagctcagct aattaagcta gcttatcgat accgtcgacc
     4321 tcgaacccca cgcccctctt taatacgacg ggcaatttgc acttcagaaa atgaagagtt
     4381 tgctttagcc ataacaaaag tccagtatgc tttttcacag cataactgga ctgatttcag
     4441 tttacaacta ttctgtctag tttaagactt tattgtcata gtttagatct attttgttca
     4501 gtttaagact ttattgtccg cccacacctg cttacgcagg gcatccattt attactcaac
     4561 cgtaaccgat tttgccaggt tacgcggctg gtcctctagc tggcgtaata gcgaagaggc
     4621 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     4681 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     4741 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     4801 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4861 gagctgcatg tgtcagaggt tttcaccgtc gtcaccgaaa cgcgcgagac gaaagggcct
     4921 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4981 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5041 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5101 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5161 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5221 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5281 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5341 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5401 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5461 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5521 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5581 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5641 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5701 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5761 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5821 tgggtcccgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5881 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5941 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6001 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6061 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6121 aaagatcaaa ggcagttatt ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6181 tttactcata tatactttag attgatttaa aacttcattt ttaatttttg cggccgcaag
     6241 atccggccac gatgcgtccg gcgtagagga tctgaagatc agcagttcaa cctgttgata
     6301 gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt actaagctct
     6361 catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc taacgtacta
     6421 agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa tataaatcag
     6481 caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata tataaggctt
     6541 ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac gcactgagaa
     6601 gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg gctgacatgg
     6661 gaattagcca tggcatcaca gtatcgtgat gacagaggca gggagtgcgg cctttttacg
     6721 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat caggggattc
     6781 cttaaggtat actttccgct gcataaccct gcttcggggt cattatagcg attttttcgg
     6841 tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc
     6901 cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg
     6961 ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc
     7021 tgcgaggctg gccgataagc tctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7081 ggtcagtgag cgaggaagcg gaagagccca aacataacag gaagaaaaat gccccgctgt
     7141 gggcggacaa aatagttggg aactgggagg ggtggaaatg gagtttttaa ggattattta
     7201 gggaagagtg acaaaataga tgggaactgg gtgtagcgtc gtaagctaat acgaaaatta
     7261 aaaatgacaa aatagtttgg aactagattt cacttatctg gttcttgagg cggttaaaag
     7321 agccgtactc ttctccgatg tcgactaggc catgatgctc attctgtggg accaaaacga
     7381 aaaaacaccc tttcgggtgt cttttctgga atttggtacc gaggctgcaa gtcccattaa
     7441 ggaggcgcgc cttattacag atcttcctca gagatgagct tctgttccag atcttcttct
     7501 gaaatcaact tttgttccag atcttcttca gagatgagtt tctgctccgc ggccgcgtca
     7561 cctcctagct gactcaaatc aatgcgtgtt tcataaagac cagtgatgga ttgatggata
     7621 agagtggcat ctaaaacttc ttttgtagac gtatatcgtt tacgatcaat tgttgtatca
     7681 aaatatttaa aagcagcggg agctccaaga ttcgtcaacg taaataaatg aataatattt
     7741 tctgcttgtt cacgtattgg tttgtctcta tgtttgttat atgcactaag aactttatct
     7801 aaattggcat ctgctaaaat aacacgctta gaaaattcac tgatttgctc aataatctca
     7861 tctaaataat gcttatgctg ctccacaaac aattgttttt gttcgttatc ttctggacta
     7921 cccttcaact tttcataatg actagctaaa tataaaaaat tcacatattt gcttggcaga
     7981 gccagctcat ttcctttttg taattctccg gcactagcca gcatccgttt acgaccgttt
     8041 tctaactcaa aaagactata tttaggtagt ttaatgatta agtctttttt aacttcctta
     8101 tatcctttag cttctaaaaa gtcaatcgga tttttttcaa aggaacttct ttccataatt
     8161 gtgatcccta gtaactcttt aacggatttt aacttcttcg atttcccttt ttccacctta
     8221 gcaaccacta ggactgaata agctaccgtt ggactatcaa aaccaccata tttttttgga
     8281 tcccagtctt ttttacgagc aataagcttg tccgaatttc tttttggtaa aattgactcc
     8341 ttggagaatc cgcctgtctg tacttctgtt ttcttgacaa tattgacttg gggcatggac
     8401 aatactttgc gcactgtggc aaaatctcgc cctttatccc agacaatttc tccagtttcc
     8461 ccattagttt cgattagagg gcgtttgcga atctctccat ttgcaagtgt aatttctgtt
     8521 ttgaagaagt tcatgatatt agagtaaaag aaatattttg cggttgcttt gcctatttct
     8581 tgctcagact tagcaatcat tttacgaaca tcataaactt tataatcacc atagacaaac
     8641 tccgattcaa gttttggata tttcttaatc aaagcagttc caacgacggc atttagatac
     8701 gcatcatggg catgatggta attgttaatc tcacgtactt tatagaattg gaaatctttt
     8761 cggaagtcag aaactaattt agattttaag gtaatcactt taacctctcg aataagttta
     8821 tcattttcat cgtatttagt attcatgcga ctatccaaaa tttgtgccac atgcttagtg
     8881 atttggcgag tttcaaccaa ttggcgtttg ataaaaccag ctttatcaag ttcactcaaa
     8941 cctccacgtt cagctttcgt taaattatca aacttacgtt gagtgattaa cttggcgttt
     9001 agaagttgtc tccaatagtt tttcatcttt ttgactactt cttcacttgg aacgttatcc
     9061 gatttaccac gatttttatc agaacgcgtt aagaccttat tgtctattga atcgtcttta
     9121 aggaaacttt gtggaacaat ggcatcgaca tcataatcac ttaaacgatt aatatctaat
     9181 tcttggtcca catacatgtc tcttccattt tggagataat agagatagag cttttcattt
     9241 tgcaattgag tattttcaac aggatgctct ttaagaatct gacttcctaa ttctttgata
     9301 ccttcttcga ttcgtttcat acgctctcgc gaatttttct ggcccttttg agttgtctga
     9361 ttttcacgtg ccatttcaat aacgatattt tctggcttat gccgccccat tactttgacc
     9421 aattcatcaa caacttttac agtctgtaaa ataccttttt taatagcagg gctaccagct
     9481 aaatttgcaa tatgttcatg taaactatcg ccttgtccag acacttgtgc tttttgaata
     9541 tcttctttaa atgtcaaact atcatcatgg atcagctgca taaaattgcg attggcaaaa
     9601 ccatctgatt tcaaaaaatc taatattgtt ttgccagatt gcttatccct aataccatta
     9661 atcaattttc gagacaaacg tccccaacca gtataacggc gacgtttaag ctgtttcatc
     9721 accttatcat caaagaggtg agcatatgtt ttaagtcttt cctcaatcat ctccctatct
     9781 tcaaataagg tcaatgttaa aacaatatcc tctaagatat cttcattttc ttcattatcc
     9841 aaaaaatctt tatctttaat aatttttagc aaatcatggt aggtacctaa tgaagcatta
     9901 aatctatctt caactcctga aatttcaaca ctatcaaaac attctatttt tttgaaataa
     9961 tcttctttta attgcttaac ggttactttt cgatttgttt tgaagagtaa atcaacaatg
    10021 gctttcttct gttcacctga aagaaatgct ggttttcgca ttccttcagt aacatatttg
    10081 acctttgtca attcgttata aaccgtaaaa tactcataaa gcaaactatg ttttggtagt
    10141 actttttcat ttggaagatt tttatcaaag tttgtcatgc gttcaataaa tgattgagct
    10201 gaagcacctt tatcgacaac ttcttcaaaa ttccatgggg taattgtttc ttcagacttc
    10261 cgagtcatcc atgcaaaacg actattgcca cgcgccaatg gaccaacata ataagggatt
    10321 cgaaaagtca agattttttc aatcttctca cgattgtctt ttaaaaatgg ataaaaatct
    10381 tcttgtcgtc tcaaaatagc atgcagctca cccaagtgaa tttgatgggg aatagagccg
    10441 ttgtcaaagg tccgttgctt gcgcagcaaa tcttcacgat ttagtttcac caataattcc
    10501 tcagtaccat ccattttttc taaaattggt ttgataaatt tataaaattc ttcttggcta
    10561 gctcccccat caatataacc tgcatatccg ttttttgatt gatcaaaaaa gatttcttta
    10621 tacttttctg gaagttgttg tcgaactaaa gcttttaaaa gagtcaagtc ttgatgatgt
    10681 tcatcgtagc gtttaatcat tgaagctgat aggggagcct tagttatttc agtatttact
    10741 cttaggatat ctgaaagtaa aatagcatct gataaattct tagctgccaa aaacaaatca
    10801 gcatattgat ctccaatttg cgccaataaa ttatctaaat catcatcgta agtatctttt
    10861 gaaagctgta atttagcatc ttctgccaaa tcaaaatttg atttaaaatt aggggtcaaa
    10921 cccaatgaca aagcaatgag attcccaaat aagccatttt tcttctcacc ggggagctga
    10981 gcaatgagat tttctaatcg tcttgattta ctcaatcgtg cagaaagaat cgctttagca
    11041 tctactccac ttgcgttaat agggttttct tcaaataatt gattgtaggt ttgtaccaac
    11101 tggataaata gtttgtccac atcactatta tcaggattta aatctccctc aatcaaaaaa
    11161 tgaccacgaa acttaatcat atgcgctaag gccaaataga ttaagcgcaa atccgcttta
    11221 tcagtagaat ctaccaattt ttttcgcaga tgatagatag ttggatattt ctcatgataa
    11281 gcaacttcat ctactatatt tccaaaaata ggatgacgtt catgcttctt atcttcttcc
    11341 accaaaaaag actcttcaag tcgatgaaag aaactatcat ctactttcgc catctcattt
    11401 gaaaaaatct cctgtagata acaaatacga ttcttccgac gtgtatacct tctacgagct
    11461 gtccgtttga gacgagtcgc ttccgctgtc tctccactgt caaataaaag agcccctata
    11521 agattttttt tgatactgtg gcggtctgta tttcccagaa ccttgaactt tttagacgga
    11581 accttatatt catcagtgat caccgcccat ccgacgctat ttgtgccgat agctaagcct
    11641 attgagtatt tcttatccat actagttttc ctgtgtgaaa cctgctgatg tgctcagtat
    11701 cttgttatcc gctcacaatg tcaatgttat ccgctcacat ttataatatt ttatctgatt
    11761 aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga
    11821 aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg
    11881 aggtaactgg cttggtgata agctgtcaaa ccagatcaat tcgcactagt ctgcagacgt
    11941 aaaaaaagcg gcgtggttag ccgctttttt aattgccgga gcgcaataaa aaagcccccg
    12001 gaaggtgatc ttccgggggc tttctcatgc gttgcttgta ttgaaaatgg gggccatgcc
    12061 tacctatctg cctcccggga agcggcatgc atttacgtt
//
