LOCUS       sequence-320852-       11307 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 11307)
  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 11307)
  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: pJMP2849
FEATURES             Location/Qualifiers
     source          1..11307
                     /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"
     RBS             303..314
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             540..1328
                     /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(1357..1404)
                     /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(1455..1541)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1644..1738)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1766..1782)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1767..1783)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2197..2216)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2316..2338
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2376..2394)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2462..2566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2567..2635,2636..3427)
                     /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(2785..2804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3728..4116
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            4385..4493
                     /note="incP origin of transfer"
     mobile_element  4597..4762
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(4917..5006)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(5016..6599,6600..6602,6603..9089,9090..9092,
                     9093..9119))
                     /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    9161..9177
                     /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        9559..9636
                     /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             9637..10719
                     /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(9656..9675)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    11004..11020
                     /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 atgcagtgat
      301 ctaaagagga gaaaggatct atgcgctcac gcaactggtc cagaaccttg accgaacgca
      361 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
      421 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
      481 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatca
      541 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
      601 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
      661 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
      721 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
      781 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
      841 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
      901 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
      961 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     1021 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     1081 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     1141 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     1201 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     1261 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     1321 tcggcaaatc tcgagcatca gcttcaaaag cgctctgaag ttcctatact ttctagagaa
     1381 taggaacttc ggaataggta cttcaagatc cccaattcga gctcatcgtc cgggccgcaa
     1441 gctcctagcg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     1501 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctcaa gctagagagt
     1561 cattacccca ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg
     1621 ccactcatcg cagtctagct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     1681 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     1741 agctcagcta attaagctag cttatcgata ccgtcgacct cgaaccccac gcccctcttt
     1801 aatacgacgg gcaatttgca cttcagaaaa tgaagagttt gctttagcca taacaaaagt
     1861 ccagtatgct ttttcacagc ataactggac tgatttcagt ttacaactat tctgtctagt
     1921 ttaagacttt attgtcatag tttagatcta ttttgttcag tttaagactt tattgtccgc
     1981 ccacacctgc ttacgcaggg catccattta ttactcaacc gtaaccgatt ttgccaggtt
     2041 acgcggctgg tcctctagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2101 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     2161 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     2221 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2281 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2341 ttcaccgtcg tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     2401 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2461 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2521 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2581 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2641 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2701 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2761 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2821 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2881 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2941 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3001 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3061 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3121 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3181 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3241 tggctggttt attgctgata aatctggagc cggtgagcgt gggtcccgcg gtatcattgc
     3301 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3361 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3421 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3481 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3541 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gcagttattg
     3601 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3661 ttgatttaaa acttcatttt taatttttgc ggccgcaaga tccggccacg atgcgtccgg
     3721 cgtagaggat ctgaagatca gcagttcaac ctgttgatag tacgtactaa gctctcatgt
     3781 ttcacgtact aagctctcat gtttaacgta ctaagctctc atgtttaacg aactaaaccc
     3841 tcatggctaa cgtactaagc tctcatggct aacgtactaa gctctcatgt ttcacgtact
     3901 aagctctcat gtttgaacaa taaaattaat ataaatcagc aacttaaata gcctctaagg
     3961 ttttaagttt tataagaaaa aaaagaatat ataaggcttt taaagctttt aaggtttaac
     4021 ggttgtggac aacaagccag ggatgtaacg cactgagaag cccttagagc ctctcaaagc
     4081 aattttgagt gacacaggaa cacttaacgg ctgacatggg aattagccat ggcatcacag
     4141 tatcgtgatg acagaggcag ggagtgcggc ctttttacgg ttcctggcct tttgctggcc
     4201 ttttgctcac atgttctttc ctgcgttatc aggggattcc ttaaggtata ctttccgctg
     4261 cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac
     4321 gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc
     4381 agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc
     4441 cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgataagct
     4501 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcgg gtcagtgagc gaggaagcgg
     4561 aagagcccaa acataacagg aagaaaaatg ccccgctgtg ggcggacaaa atagttggga
     4621 actgggaggg gtggaaatgg agtttttaag gattatttag ggaagagtga caaaatagat
     4681 gggaactggg tgtagcgtcg taagctaata cgaaaattaa aaatgacaaa atagtttgga
     4741 actagatttc acttatctgg ttcttgaggc ggttaaaaga gccgtactct tctccgatgt
     4801 cgactaggcc atgatgctca ttctgtggga ccaaaacgaa aaaacaccct ttcgggtgtc
     4861 ttttctggaa tttggtaccg aggctgcaag tcccattaag gaggcgcgcc ttattacaga
     4921 tcttcctcag agatgagctt ctgttccaga tcttcttctg aaatcaactt ttgttccaga
     4981 tcttcttcag agatgagttt ctgctccgcg gccgcgtcac ctcctagctg actcaaatca
     5041 atgcgtgttt cataaagacc agtgatggat tgatggataa gagtggcatc taaaacttct
     5101 tttgtagacg tatatcgttt acgatcaatt gttgtatcaa aatatttaaa agcagcggga
     5161 gctccaagat tcgtcaacgt aaataaatga ataatatttt ctgcttgttc acgtattggt
     5221 ttgtctctat gtttgttata tgcactaaga actttatcta aattggcatc tgctaaaata
     5281 acacgcttag aaaattcact gatttgctca ataatctcat ctaaataatg cttatgctgc
     5341 tccacaaaca attgtttttg ttcgttatct tctggactac ccttcaactt ttcataatga
     5401 ctagctaaat ataaaaaatt cacatatttg cttggcagag ccagctcatt tcctttttgt
     5461 aattctccgg cactagccag catccgttta cgaccgtttt ctaactcaaa aagactatat
     5521 ttaggtagtt taatgattaa gtctttttta acttccttat atcctttagc ttctaaaaag
     5581 tcaatcggat ttttttcaaa ggaacttctt tccataattg tgatccctag taactcttta
     5641 acggatttta acttcttcga tttccctttt tccaccttag caaccactag gactgaataa
     5701 gctaccgttg gactatcaaa accaccatat ttttttggat cccagtcttt tttacgagca
     5761 ataagcttgt ccgaatttct ttttggtaaa attgactcct tggagaatcc gcctgtctgt
     5821 acttctgttt tcttgacaat attgacttgg ggcatggaca atactttgcg cactgtggca
     5881 aaatctcgcc ctttatccca gacaatttct ccagtttccc cattagtttc gattagaggg
     5941 cgtttgcgaa tctctccatt tgcaagtgta atttctgttt tgaagaagtt catgatatta
     6001 gagtaaaaga aatattttgc ggttgctttg cctatttctt gctcagactt agcaatcatt
     6061 ttacgaacat cataaacttt ataatcacca tagacaaact ccgattcaag ttttggatat
     6121 ttcttaatca aagcagttcc aacgacggca tttagatacg catcatgggc atgatggtaa
     6181 ttgttaatct cacgtacttt atagaattgg aaatcttttc ggaagtcaga aactaattta
     6241 gattttaagg taatcacttt aacctctcga ataagtttat cattttcatc gtatttagta
     6301 ttcatgcgac tatccaaaat ttgtgccaca tgcttagtga tttggcgagt ttcaaccaat
     6361 tggcgtttga taaaaccagc tttatcaagt tcactcaaac ctccacgttc agctttcgtt
     6421 aaattatcaa acttacgttg agtgattaac ttggcgttta gaagttgtct ccaatagttt
     6481 ttcatctttt tgactacttc ttcacttgga acgttatccg atttaccacg atttttatca
     6541 gaacgcgtta agaccttatt gtctattgaa tcgtctttaa ggaaactttg tggaacaatg
     6601 gcatcgacat cataatcact taaacgatta atatctaatt cttggtccac atacatgtct
     6661 cttccatttt ggagataata gagatagagc ttttcatttt gcaattgagt attttcaaca
     6721 ggatgctctt taagaatctg acttcctaat tctttgatac cttcttcgat tcgtttcata
     6781 cgctctcgcg aatttttctg gcccttttga gttgtctgat tttcacgtgc catttcaata
     6841 acgatatttt ctggcttatg ccgccccatt actttgacca attcatcaac aacttttaca
     6901 gtctgtaaaa tacctttttt aatagcaggg ctaccagcta aatttgcaat atgttcatgt
     6961 aaactatcgc cttgtccaga cacttgtgct ttttgaatat cttctttaaa tgtcaaacta
     7021 tcatcatgga tcagctgcat aaaattgcga ttggcaaaac catctgattt caaaaaatct
     7081 aatattgttt tgccagattg cttatcccta ataccattaa tcaattttcg agacaaacgt
     7141 ccccaaccag tataacggcg acgtttaagc tgtttcatca ccttatcatc aaagaggtga
     7201 gcatatgttt taagtctttc ctcaatcatc tccctatctt caaataaggt caatgttaaa
     7261 acaatatcct ctaagatatc ttcattttct tcattatcca aaaaatcttt atctttaata
     7321 atttttagca aatcatggta ggtacctaat gaagcattaa atctatcttc aactcctgaa
     7381 atttcaacac tatcaaaaca ttctattttt ttgaaataat cttcttttaa ttgcttaacg
     7441 gttacttttc gatttgtttt gaagagtaaa tcaacaatgg ctttcttctg ttcacctgaa
     7501 agaaatgctg gttttcgcat tccttcagta acatatttga cctttgtcaa ttcgttataa
     7561 accgtaaaat actcataaag caaactatgt tttggtagta ctttttcatt tggaagattt
     7621 ttatcaaagt ttgtcatgcg ttcaataaat gattgagctg aagcaccttt atcgacaact
     7681 tcttcaaaat tccatggggt aattgtttct tcagacttcc gagtcatcca tgcaaaacga
     7741 ctattgccac gcgccaatgg accaacataa taagggattc gaaaagtcaa gattttttca
     7801 atcttctcac gattgtcttt taaaaatgga taaaaatctt cttgtcgtct caaaatagca
     7861 tgcagctcac ccaagtgaat ttgatgggga atagagccgt tgtcaaaggt ccgttgcttg
     7921 cgcagcaaat cttcacgatt tagtttcacc aataattcct cagtaccatc cattttttct
     7981 aaaattggtt tgataaattt ataaaattct tcttggctag ctcccccatc aatataacct
     8041 gcatatccgt tttttgattg atcaaaaaag atttctttat acttttctgg aagttgttgt
     8101 cgaactaaag cttttaaaag agtcaagtct tgatgatgtt catcgtagcg tttaatcatt
     8161 gaagctgata ggggagcctt agttatttca gtatttactc ttaggatatc tgaaagtaaa
     8221 atagcatctg ataaattctt agctgccaaa aacaaatcag catattgatc tccaatttgc
     8281 gccaataaat tatctaaatc atcatcgtaa gtatcttttg aaagctgtaa tttagcatct
     8341 tctgccaaat caaaatttga tttaaaatta ggggtcaaac ccaatgacaa agcaatgaga
     8401 ttcccaaata agccattttt cttctcaccg gggagctgag caatgagatt ttctaatcgt
     8461 cttgatttac tcaatcgtgc agaaagaatc gctttagcat ctactccact tgcgttaata
     8521 gggttttctt caaataattg attgtaggtt tgtaccaact ggataaatag tttgtccaca
     8581 tcactattat caggatttaa atctccctca atcaaaaaat gaccacgaaa cttaatcata
     8641 tgcgctaagg ccaaatagat taagcgcaaa tccgctttat cagtagaatc taccaatttt
     8701 tttcgcagat gatagatagt tggatatttc tcatgataag caacttcatc tactatattt
     8761 ccaaaaatag gatgacgttc atgcttctta tcttcttcca ccaaaaaaga ctcttcaagt
     8821 cgatgaaaga aactatcatc tactttcgcc atctcatttg aaaaaatctc ctgtagataa
     8881 caaatacgat tcttccgacg tgtatacctt ctacgagctg tccgtttgag acgagtcgct
     8941 tccgctgtct ctccactgtc aaataaaaga gcccctataa gatttttttt gatactgtgg
     9001 cggtctgtat ttcccagaac cttgaacttt ttagacggaa ccttatattc atcagtgatc
     9061 accgcccatc cgacgctatt tgtgccgata gctaagccta ttgagtattt cttatccata
     9121 ctagttttcc tgtgtgaaac ctgctgatgt gctcagtatc ttgttatccg ctcacaatgt
     9181 caatgttatc cgctcacatt tataatattt tatctgatta ataagatgat cttcttgaga
     9241 tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg
     9301 tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc ttggtgataa
     9361 gctgtcaaac cagatcaatt cgcactagtc tgcagacgta aaaaaagcgg cgtggttagc
     9421 cgctttttta attgccggag cgcaataaaa aagcccccgg aaggtgatct tccgggggct
     9481 ttctcatgcg ttgcttgtat tgaaaatggg ggccatgcct acctatctgc ctcccgggaa
     9541 gcggcatgca tttacgttga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga
     9601 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
     9661 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     9721 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     9781 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     9841 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     9901 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     9961 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
    10021 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
    10081 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga tggtacgcga
    10141 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
    10201 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
    10261 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
    10321 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
    10381 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
    10441 tcggtagtgg gatacgacga taccgaagat agctcatgtt atatcccgcc gttaaccacc
    10501 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
    10561 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
    10621 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
    10681 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag
    10741 ttagctcact cattaggcac cccacccggg tccttgagcc ccattatcac ctccaaagcc
    10801 ttctcgtctg gtcagtttca cctgttttac gtaaaaaccc gcttcggcgg gtttttactt
    10861 ttggggaaac acagaaaaaa gcccgcacct gacagtgcgg gctttttttt tcgaccaaag
    10921 gtgcgactac tcttgcctac tacctatcga ctgagctgaa agaattccta aagatctata
    10981 aatgtgagcg gataacattg acattgtgag cggataacaa gatacttcta gttgagacca
    11041 actttggtct ccaccatagc ggtcggtctc tgtttaagag ctatgctgga aacagcatag
    11101 caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
    11161 ttttttgaat tcatgtggct gaccgttctg ttgtctctcg ctcttccgag tagacgaaca
    11221 ataaggcctc cctaacgggg ggcctttttt attgataaca aaagtcagtg cttccgctat
    11281 ttccaaaata ccgggctaat acggttt
//
