LOCUS       sequence-317543-       11017 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 11017)
  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 11017)
  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: pJMP2844
FEATURES             Location/Qualifiers
     source          1..11017
                     /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             370..1029
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(436..455)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(1067..1114)
                     /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(1165..1251)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1354..1448)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1476..1492)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1477..1493)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1907..1926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2026..2048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2086..2104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2172..2276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2277..2345,2346..3137)
                     /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(2495..2514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3438..3826
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            4095..4203
                     /note="incP origin of transfer"
     mobile_element  4307..4472
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(4627..4716)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(4726..6309,6310..6312,6313..8799,8800..8802,
                     8803..8829))
                     /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    8871..8887
                     /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        9269..9346
                     /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             9347..10429
                     /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(9366..9385)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    10714..10730
                     /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 catcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
      361 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
      421 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
      481 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
      541 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggagttccg tatggcaatg
      601 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
      661 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
      721 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
      781 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
      841 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
      901 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
      961 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     1021 ggggcgtaat cgactggaac tcgagcatca gcttcaaaag cgctctgaag ttcctatact
     1081 ttctagagaa taggaacttc ggaataggta cttcaagatc cccaattcga gctcatcgtc
     1141 cgggccgcaa gctcctagcg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     1201 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctcaa
     1261 gctagagagt cattacccca ggcgtttaag ggcaccaata actgccttaa aaaaattacg
     1321 ccccgccctg ccactcatcg cagtctagct tggattctca ccaataaaaa acgcccggcg
     1381 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     1441 caggagtcca agctcagcta attaagctag cttatcgata ccgtcgacct cgaaccccac
     1501 gcccctcttt aatacgacgg gcaatttgca cttcagaaaa tgaagagttt gctttagcca
     1561 taacaaaagt ccagtatgct ttttcacagc ataactggac tgatttcagt ttacaactat
     1621 tctgtctagt ttaagacttt attgtcatag tttagatcta ttttgttcag tttaagactt
     1681 tattgtccgc ccacacctgc ttacgcaggg catccattta ttactcaacc gtaaccgatt
     1741 ttgccaggtt acgcggctgg tcctctagct ggcgtaatag cgaagaggcc cgcaccgatc
     1801 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     1861 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     1921 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     1981 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2041 gtcagaggtt ttcaccgtcg tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     2101 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2161 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2221 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2281 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2341 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2401 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2461 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2521 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2581 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2641 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2701 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2761 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2821 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2881 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2941 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtcccgcg
     3001 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3061 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3121 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3181 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3241 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3301 gcagttattg gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3361 atactttaga ttgatttaaa acttcatttt taatttttgc ggccgcaaga tccggccacg
     3421 atgcgtccgg cgtagaggat ctgaagatca gcagttcaac ctgttgatag tacgtactaa
     3481 gctctcatgt ttcacgtact aagctctcat gtttaacgta ctaagctctc atgtttaacg
     3541 aactaaaccc tcatggctaa cgtactaagc tctcatggct aacgtactaa gctctcatgt
     3601 ttcacgtact aagctctcat gtttgaacaa taaaattaat ataaatcagc aacttaaata
     3661 gcctctaagg ttttaagttt tataagaaaa aaaagaatat ataaggcttt taaagctttt
     3721 aaggtttaac ggttgtggac aacaagccag ggatgtaacg cactgagaag cccttagagc
     3781 ctctcaaagc aattttgagt gacacaggaa cacttaacgg ctgacatggg aattagccat
     3841 ggcatcacag tatcgtgatg acagaggcag ggagtgcggc ctttttacgg ttcctggcct
     3901 tttgctggcc ttttgctcac atgttctttc ctgcgttatc aggggattcc ttaaggtata
     3961 ctttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc
     4021 tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc
     4081 caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc
     4141 ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg
     4201 ccgataagct ctgataccgc tcgccgcagc cgaacgaccg agcgcagcgg gtcagtgagc
     4261 gaggaagcgg aagagcccaa acataacagg aagaaaaatg ccccgctgtg ggcggacaaa
     4321 atagttggga actgggaggg gtggaaatgg agtttttaag gattatttag ggaagagtga
     4381 caaaatagat gggaactggg tgtagcgtcg taagctaata cgaaaattaa aaatgacaaa
     4441 atagtttgga actagatttc acttatctgg ttcttgaggc ggttaaaaga gccgtactct
     4501 tctccgatgt cgactaggcc atgatgctca ttctgtggga ccaaaacgaa aaaacaccct
     4561 ttcgggtgtc ttttctggaa tttggtaccg aggctgcaag tcccattaag gaggcgcgcc
     4621 ttattacaga tcttcctcag agatgagctt ctgttccaga tcttcttctg aaatcaactt
     4681 ttgttccaga tcttcttcag agatgagttt ctgctccgcg gccgcgtcac ctcctagctg
     4741 actcaaatca atgcgtgttt cataaagacc agtgatggat tgatggataa gagtggcatc
     4801 taaaacttct tttgtagacg tatatcgttt acgatcaatt gttgtatcaa aatatttaaa
     4861 agcagcggga gctccaagat tcgtcaacgt aaataaatga ataatatttt ctgcttgttc
     4921 acgtattggt ttgtctctat gtttgttata tgcactaaga actttatcta aattggcatc
     4981 tgctaaaata acacgcttag aaaattcact gatttgctca ataatctcat ctaaataatg
     5041 cttatgctgc tccacaaaca attgtttttg ttcgttatct tctggactac ccttcaactt
     5101 ttcataatga ctagctaaat ataaaaaatt cacatatttg cttggcagag ccagctcatt
     5161 tcctttttgt aattctccgg cactagccag catccgttta cgaccgtttt ctaactcaaa
     5221 aagactatat ttaggtagtt taatgattaa gtctttttta acttccttat atcctttagc
     5281 ttctaaaaag tcaatcggat ttttttcaaa ggaacttctt tccataattg tgatccctag
     5341 taactcttta acggatttta acttcttcga tttccctttt tccaccttag caaccactag
     5401 gactgaataa gctaccgttg gactatcaaa accaccatat ttttttggat cccagtcttt
     5461 tttacgagca ataagcttgt ccgaatttct ttttggtaaa attgactcct tggagaatcc
     5521 gcctgtctgt acttctgttt tcttgacaat attgacttgg ggcatggaca atactttgcg
     5581 cactgtggca aaatctcgcc ctttatccca gacaatttct ccagtttccc cattagtttc
     5641 gattagaggg cgtttgcgaa tctctccatt tgcaagtgta atttctgttt tgaagaagtt
     5701 catgatatta gagtaaaaga aatattttgc ggttgctttg cctatttctt gctcagactt
     5761 agcaatcatt ttacgaacat cataaacttt ataatcacca tagacaaact ccgattcaag
     5821 ttttggatat ttcttaatca aagcagttcc aacgacggca tttagatacg catcatgggc
     5881 atgatggtaa ttgttaatct cacgtacttt atagaattgg aaatcttttc ggaagtcaga
     5941 aactaattta gattttaagg taatcacttt aacctctcga ataagtttat cattttcatc
     6001 gtatttagta ttcatgcgac tatccaaaat ttgtgccaca tgcttagtga tttggcgagt
     6061 ttcaaccaat tggcgtttga taaaaccagc tttatcaagt tcactcaaac ctccacgttc
     6121 agctttcgtt aaattatcaa acttacgttg agtgattaac ttggcgttta gaagttgtct
     6181 ccaatagttt ttcatctttt tgactacttc ttcacttgga acgttatccg atttaccacg
     6241 atttttatca gaacgcgtta agaccttatt gtctattgaa tcgtctttaa ggaaactttg
     6301 tggaacaatg gcatcgacat cataatcact taaacgatta atatctaatt cttggtccac
     6361 atacatgtct cttccatttt ggagataata gagatagagc ttttcatttt gcaattgagt
     6421 attttcaaca ggatgctctt taagaatctg acttcctaat tctttgatac cttcttcgat
     6481 tcgtttcata cgctctcgcg aatttttctg gcccttttga gttgtctgat tttcacgtgc
     6541 catttcaata acgatatttt ctggcttatg ccgccccatt actttgacca attcatcaac
     6601 aacttttaca gtctgtaaaa tacctttttt aatagcaggg ctaccagcta aatttgcaat
     6661 atgttcatgt aaactatcgc cttgtccaga cacttgtgct ttttgaatat cttctttaaa
     6721 tgtcaaacta tcatcatgga tcagctgcat aaaattgcga ttggcaaaac catctgattt
     6781 caaaaaatct aatattgttt tgccagattg cttatcccta ataccattaa tcaattttcg
     6841 agacaaacgt ccccaaccag tataacggcg acgtttaagc tgtttcatca ccttatcatc
     6901 aaagaggtga gcatatgttt taagtctttc ctcaatcatc tccctatctt caaataaggt
     6961 caatgttaaa acaatatcct ctaagatatc ttcattttct tcattatcca aaaaatcttt
     7021 atctttaata atttttagca aatcatggta ggtacctaat gaagcattaa atctatcttc
     7081 aactcctgaa atttcaacac tatcaaaaca ttctattttt ttgaaataat cttcttttaa
     7141 ttgcttaacg gttacttttc gatttgtttt gaagagtaaa tcaacaatgg ctttcttctg
     7201 ttcacctgaa agaaatgctg gttttcgcat tccttcagta acatatttga cctttgtcaa
     7261 ttcgttataa accgtaaaat actcataaag caaactatgt tttggtagta ctttttcatt
     7321 tggaagattt ttatcaaagt ttgtcatgcg ttcaataaat gattgagctg aagcaccttt
     7381 atcgacaact tcttcaaaat tccatggggt aattgtttct tcagacttcc gagtcatcca
     7441 tgcaaaacga ctattgccac gcgccaatgg accaacataa taagggattc gaaaagtcaa
     7501 gattttttca atcttctcac gattgtcttt taaaaatgga taaaaatctt cttgtcgtct
     7561 caaaatagca tgcagctcac ccaagtgaat ttgatgggga atagagccgt tgtcaaaggt
     7621 ccgttgcttg cgcagcaaat cttcacgatt tagtttcacc aataattcct cagtaccatc
     7681 cattttttct aaaattggtt tgataaattt ataaaattct tcttggctag ctcccccatc
     7741 aatataacct gcatatccgt tttttgattg atcaaaaaag atttctttat acttttctgg
     7801 aagttgttgt cgaactaaag cttttaaaag agtcaagtct tgatgatgtt catcgtagcg
     7861 tttaatcatt gaagctgata ggggagcctt agttatttca gtatttactc ttaggatatc
     7921 tgaaagtaaa atagcatctg ataaattctt agctgccaaa aacaaatcag catattgatc
     7981 tccaatttgc gccaataaat tatctaaatc atcatcgtaa gtatcttttg aaagctgtaa
     8041 tttagcatct tctgccaaat caaaatttga tttaaaatta ggggtcaaac ccaatgacaa
     8101 agcaatgaga ttcccaaata agccattttt cttctcaccg gggagctgag caatgagatt
     8161 ttctaatcgt cttgatttac tcaatcgtgc agaaagaatc gctttagcat ctactccact
     8221 tgcgttaata gggttttctt caaataattg attgtaggtt tgtaccaact ggataaatag
     8281 tttgtccaca tcactattat caggatttaa atctccctca atcaaaaaat gaccacgaaa
     8341 cttaatcata tgcgctaagg ccaaatagat taagcgcaaa tccgctttat cagtagaatc
     8401 taccaatttt tttcgcagat gatagatagt tggatatttc tcatgataag caacttcatc
     8461 tactatattt ccaaaaatag gatgacgttc atgcttctta tcttcttcca ccaaaaaaga
     8521 ctcttcaagt cgatgaaaga aactatcatc tactttcgcc atctcatttg aaaaaatctc
     8581 ctgtagataa caaatacgat tcttccgacg tgtatacctt ctacgagctg tccgtttgag
     8641 acgagtcgct tccgctgtct ctccactgtc aaataaaaga gcccctataa gatttttttt
     8701 gatactgtgg cggtctgtat ttcccagaac cttgaacttt ttagacggaa ccttatattc
     8761 atcagtgatc accgcccatc cgacgctatt tgtgccgata gctaagccta ttgagtattt
     8821 cttatccata ctagttttcc tgtgtgaaac ctgctgatgt gctcagtatc ttgttatccg
     8881 ctcacaatgt caatgttatc cgctcacatt tataatattt tatctgatta ataagatgat
     8941 cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct
     9001 tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc
     9061 ttggtgataa gctgtcaaac cagatcaatt cgcactagtc tgcagacgta aaaaaagcgg
     9121 cgtggttagc cgctttttta attgccggag cgcaataaaa aagcccccgg aaggtgatct
     9181 tccgggggct ttctcatgcg ttgcttgtat tgaaaatggg ggccatgcct acctatctgc
     9241 ctcccgggaa gcggcatgca tttacgttga caccatcgaa tggtgcaaaa cctttcgcgg
     9301 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     9361 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     9421 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     9481 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     9541 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     9601 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     9661 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     9721 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     9781 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     9841 tggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     9901 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     9961 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
    10021 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
    10081 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
    10141 tgcggatatc tcggtagtgg gatacgacga taccgaagat agctcatgtt atatcccgcc
    10201 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
    10261 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
    10321 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
    10381 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
    10441 ttaatgtaag ttagctcact cattaggcac cccacccggg tccttgagcc ccattatcac
    10501 ctccaaagcc ttctcgtctg gtcagtttca cctgttttac gtaaaaaccc gcttcggcgg
    10561 gtttttactt ttggggaaac acagaaaaaa gcccgcacct gacagtgcgg gctttttttt
    10621 tcgaccaaag gtgcgactac tcttgcctac tacctatcga ctgagctgaa agaattccta
    10681 aagatctata aatgtgagcg gataacattg acattgtgag cggataacaa gatacttcta
    10741 gttgagacca actttggtct ccaccatagc ggtcggtctc tgtttaagag ctatgctgga
    10801 aacagcatag caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
    10861 tcggtgcttt ttttttgaat tcatgtggct gaccgttctg ttgtctctcg ctcttccgag
    10921 tagacgaaca ataaggcctc cctaacgggg ggcctttttt attgataaca aaagtcagtg
    10981 cttccgctat ttccaaaata ccgggctaat acggttt
//
