LOCUS       sequence-317545-       11194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobile CRISPRi empty sgRNA (BsaI) for cloning new sgRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11194)
  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 11194)
  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: pJMP2846
FEATURES             Location/Qualifiers
     source          1..11194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(135..182)
                     /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(259..264,265..281,282..287)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             398..1192
                     /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(452..471)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1062..1081
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(1244..1291)
                     /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(1342..1428)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1531..1625)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1653..1669)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1654..1670)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2084..2103)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2203..2225
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2263..2281)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2349..2453
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2454..2522,2523..3314)
                     /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(2672..2691)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3615..4003
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            4272..4380
                     /note="incP origin of transfer"
     mobile_element  4484..4649
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(4804..4893)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(4903..6486,6487..6489,6490..8976,8977..8979,
                     8980..9006))
                     /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    9048..9064
                     /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        9446..9523
                     /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             9524..10606
                     /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(9543..9562)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    10891..10907
                     /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 aaacgacctc ctggatttgc tcagacagcc ttttcgtcat tcgtttcagc caaaaaactt
       61 aagaccgccg gtcttgtcca ctaccttgca gtaatgcggt ggacaggatc ggcggttttc
      121 ttttctcttc tcaagaagtt cctatacttt ctagagaata ggaacttcgg aataggaact
      181 tcctcctgaa cggccataag aacgaaggct gtctgttgaa ctctcgagcc gtggaaacgg
      241 atgaaggcac gaacccagtt gacataagcc tgttcggttc gtaaactgta atgcaagtag
      301 cgtatgcgct caaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
      361 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      421 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      481 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      541 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      601 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      661 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      721 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      781 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      841 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      901 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
      961 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1021 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1081 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     1141 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1201 ctggggttcg aaatgactcg agcatcagct tcaaaagcgc tctgaagttc ctatactttc
     1261 tagagaatag gaacttcgga ataggtactt caagatcccc aattcgagct catcgtccgg
     1321 gccgcaagct cctagcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     1381 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctcaagct
     1441 agagagtcat taccccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
     1501 cgccctgcca ctcatcgcag tctagcttgg attctcacca ataaaaaacg cccggcggca
     1561 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     1621 gagtccaagc tcagctaatt aagctagctt atcgataccg tcgacctcga accccacgcc
     1681 cctctttaat acgacgggca atttgcactt cagaaaatga agagtttgct ttagccataa
     1741 caaaagtcca gtatgctttt tcacagcata actggactga tttcagttta caactattct
     1801 gtctagttta agactttatt gtcatagttt agatctattt tgttcagttt aagactttat
     1861 tgtccgccca cacctgctta cgcagggcat ccatttatta ctcaaccgta accgattttg
     1921 ccaggttacg cggctggtcc tctagctggc gtaatagcga agaggcccgc accgatcgcc
     1981 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     2041 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     2101 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2161 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2221 agaggttttc accgtcgtca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     2281 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2341 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2401 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2461 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2521 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2581 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2641 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2701 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2761 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2821 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2881 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2941 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3001 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3061 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3121 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcccgcggta
     3181 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3241 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3301 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3361 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3421 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggca
     3481 gttattggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3541 ctttagattg atttaaaact tcatttttaa tttttgcggc cgcaagatcc ggccacgatg
     3601 cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac gtactaagct
     3661 ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg tttaacgaac
     3721 taaaccctca tggctaacgt actaagctct catggctaac gtactaagct ctcatgtttc
     3781 acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac ttaaatagcc
     3841 tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa agcttttaag
     3901 gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc ttagagcctc
     3961 tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat tagccatggc
     4021 atcacagtat cgtgatgaca gaggcaggga gtgcggcctt tttacggttc ctggcctttt
     4081 gctggccttt tgctcacatg ttctttcctg cgttatcagg ggattcctta aggtatactt
     4141 tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt
     4201 ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa
     4261 cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc
     4321 actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg
     4381 ataagctctg ataccgctcg ccgcagccga acgaccgagc gcagcgggtc agtgagcgag
     4441 gaagcggaag agcccaaaca taacaggaag aaaaatgccc cgctgtgggc ggacaaaata
     4501 gttgggaact gggaggggtg gaaatggagt ttttaaggat tatttaggga agagtgacaa
     4561 aatagatggg aactgggtgt agcgtcgtaa gctaatacga aaattaaaaa tgacaaaata
     4621 gtttggaact agatttcact tatctggttc ttgaggcggt taaaagagcc gtactcttct
     4681 ccgatgtcga ctaggccatg atgctcattc tgtgggacca aaacgaaaaa acaccctttc
     4741 gggtgtcttt tctggaattt ggtaccgagg ctgcaagtcc cattaaggag gcgcgcctta
     4801 ttacagatct tcctcagaga tgagcttctg ttccagatct tcttctgaaa tcaacttttg
     4861 ttccagatct tcttcagaga tgagtttctg ctccgcggcc gcgtcacctc ctagctgact
     4921 caaatcaatg cgtgtttcat aaagaccagt gatggattga tggataagag tggcatctaa
     4981 aacttctttt gtagacgtat atcgtttacg atcaattgtt gtatcaaaat atttaaaagc
     5041 agcgggagct ccaagattcg tcaacgtaaa taaatgaata atattttctg cttgttcacg
     5101 tattggtttg tctctatgtt tgttatatgc actaagaact ttatctaaat tggcatctgc
     5161 taaaataaca cgcttagaaa attcactgat ttgctcaata atctcatcta aataatgctt
     5221 atgctgctcc acaaacaatt gtttttgttc gttatcttct ggactaccct tcaacttttc
     5281 ataatgacta gctaaatata aaaaattcac atatttgctt ggcagagcca gctcatttcc
     5341 tttttgtaat tctccggcac tagccagcat ccgtttacga ccgttttcta actcaaaaag
     5401 actatattta ggtagtttaa tgattaagtc ttttttaact tccttatatc ctttagcttc
     5461 taaaaagtca atcggatttt tttcaaagga acttctttcc ataattgtga tccctagtaa
     5521 ctctttaacg gattttaact tcttcgattt ccctttttcc accttagcaa ccactaggac
     5581 tgaataagct accgttggac tatcaaaacc accatatttt tttggatccc agtctttttt
     5641 acgagcaata agcttgtccg aatttctttt tggtaaaatt gactccttgg agaatccgcc
     5701 tgtctgtact tctgttttct tgacaatatt gacttggggc atggacaata ctttgcgcac
     5761 tgtggcaaaa tctcgccctt tatcccagac aatttctcca gtttccccat tagtttcgat
     5821 tagagggcgt ttgcgaatct ctccatttgc aagtgtaatt tctgttttga agaagttcat
     5881 gatattagag taaaagaaat attttgcggt tgctttgcct atttcttgct cagacttagc
     5941 aatcatttta cgaacatcat aaactttata atcaccatag acaaactccg attcaagttt
     6001 tggatatttc ttaatcaaag cagttccaac gacggcattt agatacgcat catgggcatg
     6061 atggtaattg ttaatctcac gtactttata gaattggaaa tcttttcgga agtcagaaac
     6121 taatttagat tttaaggtaa tcactttaac ctctcgaata agtttatcat tttcatcgta
     6181 tttagtattc atgcgactat ccaaaatttg tgccacatgc ttagtgattt ggcgagtttc
     6241 aaccaattgg cgtttgataa aaccagcttt atcaagttca ctcaaacctc cacgttcagc
     6301 tttcgttaaa ttatcaaact tacgttgagt gattaacttg gcgtttagaa gttgtctcca
     6361 atagtttttc atctttttga ctacttcttc acttggaacg ttatccgatt taccacgatt
     6421 tttatcagaa cgcgttaaga ccttattgtc tattgaatcg tctttaagga aactttgtgg
     6481 aacaatggca tcgacatcat aatcacttaa acgattaata tctaattctt ggtccacata
     6541 catgtctctt ccattttgga gataatagag atagagcttt tcattttgca attgagtatt
     6601 ttcaacagga tgctctttaa gaatctgact tcctaattct ttgatacctt cttcgattcg
     6661 tttcatacgc tctcgcgaat ttttctggcc cttttgagtt gtctgatttt cacgtgccat
     6721 ttcaataacg atattttctg gcttatgccg ccccattact ttgaccaatt catcaacaac
     6781 ttttacagtc tgtaaaatac cttttttaat agcagggcta ccagctaaat ttgcaatatg
     6841 ttcatgtaaa ctatcgcctt gtccagacac ttgtgctttt tgaatatctt ctttaaatgt
     6901 caaactatca tcatggatca gctgcataaa attgcgattg gcaaaaccat ctgatttcaa
     6961 aaaatctaat attgttttgc cagattgctt atccctaata ccattaatca attttcgaga
     7021 caaacgtccc caaccagtat aacggcgacg tttaagctgt ttcatcacct tatcatcaaa
     7081 gaggtgagca tatgttttaa gtctttcctc aatcatctcc ctatcttcaa ataaggtcaa
     7141 tgttaaaaca atatcctcta agatatcttc attttcttca ttatccaaaa aatctttatc
     7201 tttaataatt tttagcaaat catggtaggt acctaatgaa gcattaaatc tatcttcaac
     7261 tcctgaaatt tcaacactat caaaacattc tatttttttg aaataatctt cttttaattg
     7321 cttaacggtt acttttcgat ttgttttgaa gagtaaatca acaatggctt tcttctgttc
     7381 acctgaaaga aatgctggtt ttcgcattcc ttcagtaaca tatttgacct ttgtcaattc
     7441 gttataaacc gtaaaatact cataaagcaa actatgtttt ggtagtactt tttcatttgg
     7501 aagattttta tcaaagtttg tcatgcgttc aataaatgat tgagctgaag cacctttatc
     7561 gacaacttct tcaaaattcc atggggtaat tgtttcttca gacttccgag tcatccatgc
     7621 aaaacgacta ttgccacgcg ccaatggacc aacataataa gggattcgaa aagtcaagat
     7681 tttttcaatc ttctcacgat tgtcttttaa aaatggataa aaatcttctt gtcgtctcaa
     7741 aatagcatgc agctcaccca agtgaatttg atggggaata gagccgttgt caaaggtccg
     7801 ttgcttgcgc agcaaatctt cacgatttag tttcaccaat aattcctcag taccatccat
     7861 tttttctaaa attggtttga taaatttata aaattcttct tggctagctc ccccatcaat
     7921 ataacctgca tatccgtttt ttgattgatc aaaaaagatt tctttatact tttctggaag
     7981 ttgttgtcga actaaagctt ttaaaagagt caagtcttga tgatgttcat cgtagcgttt
     8041 aatcattgaa gctgataggg gagccttagt tatttcagta tttactctta ggatatctga
     8101 aagtaaaata gcatctgata aattcttagc tgccaaaaac aaatcagcat attgatctcc
     8161 aatttgcgcc aataaattat ctaaatcatc atcgtaagta tcttttgaaa gctgtaattt
     8221 agcatcttct gccaaatcaa aatttgattt aaaattaggg gtcaaaccca atgacaaagc
     8281 aatgagattc ccaaataagc catttttctt ctcaccgggg agctgagcaa tgagattttc
     8341 taatcgtctt gatttactca atcgtgcaga aagaatcgct ttagcatcta ctccacttgc
     8401 gttaataggg ttttcttcaa ataattgatt gtaggtttgt accaactgga taaatagttt
     8461 gtccacatca ctattatcag gatttaaatc tccctcaatc aaaaaatgac cacgaaactt
     8521 aatcatatgc gctaaggcca aatagattaa gcgcaaatcc gctttatcag tagaatctac
     8581 caattttttt cgcagatgat agatagttgg atatttctca tgataagcaa cttcatctac
     8641 tatatttcca aaaataggat gacgttcatg cttcttatct tcttccacca aaaaagactc
     8701 ttcaagtcga tgaaagaaac tatcatctac tttcgccatc tcatttgaaa aaatctcctg
     8761 tagataacaa atacgattct tccgacgtgt ataccttcta cgagctgtcc gtttgagacg
     8821 agtcgcttcc gctgtctctc cactgtcaaa taaaagagcc cctataagat tttttttgat
     8881 actgtggcgg tctgtatttc ccagaacctt gaacttttta gacggaacct tatattcatc
     8941 agtgatcacc gcccatccga cgctatttgt gccgatagct aagcctattg agtatttctt
     9001 atccatacta gttttcctgt gtgaaacctg ctgatgtgct cagtatcttg ttatccgctc
     9061 acaatgtcaa tgttatccgc tcacatttat aatattttat ctgattaata agatgatctt
     9121 cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc
     9181 agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg
     9241 gtgataagct gtcaaaccag atcaattcgc actagtctgc agacgtaaaa aaagcggcgt
     9301 ggttagccgc ttttttaatt gccggagcgc aataaaaaag cccccggaag gtgatcttcc
     9361 gggggctttc tcatgcgttg cttgtattga aaatgggggc catgcctacc tatctgcctc
     9421 ccgggaagcg gcatgcattt acgttgacac catcgaatgg tgcaaaacct ttcgcggtat
     9481 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     9541 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     9601 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     9661 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     9721 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     9781 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     9841 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     9901 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     9961 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagatgg
    10021 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
    10081 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
    10141 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
    10201 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
    10261 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
    10321 ggatatctcg gtagtgggat acgacgatac cgaagatagc tcatgttata tcccgccgtt
    10381 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
    10441 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
    10501 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
    10561 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
    10621 atgtaagtta gctcactcat taggcacccc acccgggtcc ttgagcccca ttatcacctc
    10681 caaagccttc tcgtctggtc agtttcacct gttttacgta aaaacccgct tcggcgggtt
    10741 tttacttttg gggaaacaca gaaaaaagcc cgcacctgac agtgcgggct ttttttttcg
    10801 accaaaggtg cgactactct tgcctactac ctatcgactg agctgaaaga attcctaaag
    10861 atctataaat gtgagcggat aacattgaca ttgtgagcgg ataacaagat acttctagtt
    10921 gagaccaact ttggtctcca ccatagcggt cggtctctgt ttaagagcta tgctggaaac
    10981 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
    11041 gtgctttttt tttgaattca tgtggctgac cgttctgttg tctctcgctc ttccgagtag
    11101 acgaacaata aggcctccct aacggggggc cttttttatt gataacaaaa gtcagtgctt
    11161 ccgctatttc caaaataccg ggctaatacg gttt
//
