LOCUS       sequence-232560-       10837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knockdown folA in P. aeruginosa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10837)
  AUTHORS   Peters JM, Koo BM, Patino R, Heussler GE, Hearne CC, Qu J, Inclan 
            YF, Hawkins JS, Lu CHS, Silvis MR, Harden MM, Osadnik H, Peters JE, 
            Engel JN, Dutton RJ, Grossman AD, Gross CA, Rosenberg OS
  TITLE     Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi.
  JOURNAL   Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 
            10.1038/s41564-018-0327-z.
  PUBMED    30617347
REFERENCE   2  (bases 1 to 10837)
  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: "Nat 
            Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 
            10.1038/s41564-018-0327-z."
COMMENT     Sequence Label: pJMP1239
FEATURES             Location/Qualifiers
     source          1..10837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  152..317
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(472..561)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(571..2154,2155..2157,2158..4644,4645..4647,
                     4648..4674))
                     /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"
     CDS             complement(571..4674)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /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"
     RBS             4686..4708
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(4735..5019)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(4793..4812)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             5046..5924
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(6307..6312,6313..6329,6330..6335)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(6732..6779)
                     /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(6896..6901,6902..6918,6919..6924)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             7113..7646
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(7694..7741)
                     /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(7792..7878)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(7981..8075)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(8103..8119)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8104..8120)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(8534..8553)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8653..8675
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8713..8731)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8799..8903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8904..8972,8973..9764)
                     /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(9122..9141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10065..10453
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            10722..10830
                     /note="incP origin of transfer"
ORIGIN
        1 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcgg gtcagtgagc gaggaagcgg
       61 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg
      121 cccaaacata acaggaagaa aaatgccccg ctgtgggcgg acaaaatagt tgggaactgg
      181 gaggggtgga aatggagttt ttaaggatta tttagggaag agtgacaaaa tagatgggaa
      241 ctgggtgtag cgtcgtaagc taatacgaaa attaaaaatg acaaaatagt ttggaactag
      301 atttcactta tctggttctt gaggcggtta aaagagccgt actcttctcc gatgtcgact
      361 aggccatgat gctcattctg tgggaccaaa acgaaaaaac accctttcgg gtgtcttttc
      421 tggaatttgg taccgaggct gcaagtccca ttaaggaggc gcgccttatt acagatcttc
      481 ctcagagatg agcttctgtt ccagatcttc ttctgaaatc aacttttgtt ccagatcttc
      541 ttcagagatg agtttctgct ccgcggccgc gtcgcctccc agctgagaca ggtcgatccg
      601 tgtctcgtac aggccggtga tgctctggtg gatcagggtg gcgtccagca cctctttggt
      661 gctggtgtac ctcttccggt cgatggtggt gtcaaagtac ttgaaggcgg caggggctcc
      721 cagattggtc agggtaaaca ggtggatgat attctcggcc tgctctctga taggcttgtc
      781 tctgtgcttg ttgtaggcgc tcagcacctt gtccagatta gcgtcggcca ggatcactct
      841 cttggagaac tcgctgatct gctcgatgat ctcgtccagg tagtgtttgt gctgttccac
      901 aaacagctgt ttctgctcat tatcctcggg ggagcccttc agcttctcat agtggctggc
      961 caggtacagg aagttcacat atttggaggg cagggccagt tcgtttccct tctgcagttc
     1021 gccggcagag gccagcattc tcttccggcc gttttccagc tcgaacaggg agtacttagg
     1081 cagcttgatg atcaggtcct ttttcacttc tttgtagccc ttggcttcca gaaagtcgat
     1141 gggattcttc tcgaagctgc ttctttccat gatggtgatc cccagcagct ctttcacact
     1201 cttcagtttc ttggacttgc ccttttccac tttggccacc accagcacag aataggccac
     1261 ggtggggctg tcgaagccgc cgtacttctt agggtcccag tccttctttc tggcgatcag
     1321 cttgtcgctg ttcctcttgg gcaggataga ctctttgctg aagccgcctg tctgcacctc
     1381 ggtctttttc acgatattca cttggggcat agacagcact ttccgcacgg tggcaaagtc
     1441 ccggccctta tcccacacga tctcgcctgt ttcgccgttt gtctcgatca gaggccgctt
     1501 ccggatctcg ccgttggcca gggtaatctc ggtcttgaaa aagttcatga tgttgctgta
     1561 gaagaagtac ttggcggtag ccttgccgat ttcctgctcg ctcttggcga tcatcttccg
     1621 cacgtcgtac accttgtagt cgccgtacac gaactcgctt tccagcttag ggtacttttt
     1681 gatcagggcg gttcccacga cggcgttcag gtaggcgtcg tgggcgtggt ggtagttgtt
     1741 gatctcgcgc actttgtaaa actggaaatc cttccggaaa tcggacacca gcttggactt
     1801 cagggtgatc actttcactt cccggatcag tttgtcgttc tcgtcgtact tagtgttcat
     1861 ccgggagtcc aggatctgtg ccacgtgctt tgtgatctgc cgggtttcca ccagctgtct
     1921 cttgatgaag ccggccttat ccagttcgct caggccgcct ctctcggcct tggtcagatt
     1981 gtcgaacttc ctctgggtaa tcagcttggc attcagcagc tggcgccagt agttcttcat
     2041 cttcttcacg acctcttcgg agggcacgtt gtcgctcttg ccccggttct tgtcgctccg
     2101 agtcagcact ttgttatcga tggagtcgtc cttcagaaag ctctgaggca cgatagcgtc
     2161 cacatcgtag tcggacagcc ggttgatgtc cagttcctgg tccacgtaca tatcccgccc
     2221 attctgcagg tagtacaggt acagcttctc gttctgcagc tgggtgtttt ccacggggtg
     2281 ttctttcagg atctggctgc ccagctcttt gatgccctct tcgatccgct tcattctctc
     2341 gcggctgttc ttctgtccct tctgggtggt ctggttctct ctggccattt cgatcacgat
     2401 gttctcgggc ttgtgccggc ccatcacttt cacgagctcg tccaccacct tcactgtctg
     2461 caggatgccc ttcttaatgg cggggctgcc ggccagattg gcaatgtgct cgtgcaggct
     2521 atcgccctgg ccggacacct gggctttctg gatgtcctct ttaaaggtca ggctgtcgtc
     2581 gtggatcagc tgcatgaagt ttctgttggc gaagccgtcg gacttcagga aatccaggat
     2641 tgtcttgccg gactgcttgt cccggatgcc gttgatcagc ttccggctca gcctgcccca
     2701 gccggtgtat ctccgccgct tcagctgctt catcactttg tcgtcgaaca ggtgggcata
     2761 ggttttcagc cgttcctcga tcatctctct gtcctcaaac agtgtcaggg tcagcacgat
     2821 atcttccaga atgtcctcgt tttcctcatt gtccaggaag tccttgtcct tgataatttt
     2881 cagcagatcg tggtatgtgc ccagggaggc gttgaaccga tcttccacgc cggagatttc
     2941 cacggagtcg aagcactcga ttttcttgaa gtagtcctct ttcagctgct tcacggtcac
     3001 tttccggttg gtcttgaaca gcaggtccac gatggctttt ttctgctcgc cgctcaggaa
     3061 ggcgggcttt ctcattccct cggtcacgta tttcactttg gtcagctcgt tgtacacggt
     3121 gaagtactcg tacagcaggc tgtgcttggg cagcaccttc tcgttgggca ggttcttatc
     3181 gaagttggtc atccgctcga tgaagctctg ggcgctggcg cccttgtcca ccacttcctc
     3241 gaagttccag ggggtgatgg tttcctcgct ctttctggtc atccaggcga atctgctgtt
     3301 tcccctggcc agagggccca cgtagtaggg gatgcggaag gtcaggatct tctcgatctt
     3361 ttcccggttg tccttcagga atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag
     3421 ctctcccagg tggatctggt gggggatgct gccgttgtcg aaggtccgct gcttccgcag
     3481 caggtcctct ctgttcagct tcacgagcag ttcctcggtg ccgtccatct tttccaggat
     3541 gggcttgatg aacttgtaga actcttcctg gctggctccg ccatcgatgt agccggcgta
     3601 gccgttcttg ctctggtcga agaaaatctc tttgtacttc tcaggcagct gctgccgcac
     3661 gagagctttc agcagggtca ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc
     3721 gctcaggggg gccttggtga tctcggtgtt cactctcagg atgtcgctca gcaggatggc
     3781 gtcggacagg ttcttggcgg ccagaaacag gtcggcgtac tggtcgccga tctgggccag
     3841 caggttgtcc aggtcgtcgt cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc
     3901 caggtcgaag ttgctcttga agttgggggt caggcccagg ctcagggcaa tcaggttgcc
     3961 gaacaggcca ttcttcttct cgccgggcag ctgggcgatc agattttcca gccgtctgct
     4021 cttgctcagt ctggcagaca ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt
     4081 ttcctcgaac agctggttgt aggtctgcac cagctggatg aacagcttgt ccacgtcgct
     4141 gttgtcgggg ttcaggtcgc cctcgatcag gaagtggccc cggaacttga tcatgtgggc
     4201 cagggccaga tagatcagcc gcaggtcggc cttgtcggtg ctgtccacca gtttctttct
     4261 caggtggtag atggtggggt acttctcgtg gtaggccacc tcgtccacga tgttgccgaa
     4321 gatggggtgc cgctcgtgct tcttatcctc ttccaccagg aaggactctt ccagtctgtg
     4381 gaagaagctg tcgtccacct tggccatctc gttgctgaag atctcttgca gatagcagat
     4441 ccggttcttc cgtctggtgt atcttcttct ggcggttctc ttcagccggg tggcctcggc
     4501 tgtttctccg ctgtcgaaca gcagggcgcc gatcaggttc ttcttgatgc tgtgccggtc
     4561 ggtgttgccc agcaccttga atttcttgct gggcaccttg tactcgtcgg tgatcacggc
     4621 ccagcccaca gagttggtgc cgatggccag gccgatgctg tacttcttgt ccatgggtat
     4681 gtatatctcc ttcttaaagt taaacaaaat tatttctagc ccaaaaaaac gggtatggag
     4741 aaacagtaga gagttgcgat aaaaagcgtc aggtaggatc cgctaatctt atggataaaa
     4801 atgctatggc atagcaaagt gtgacgccgt gcaaataatc aatgtggact tttctgccgt
     4861 gattatagac acttttgtta cgcgtttttg tcatggcttt ggtcccgctt tgttacagaa
     4921 tgcttttaat aagcggggtt accggtttgg ttagcgagaa gagccagtaa aagacgcagt
     4981 gacggcaatg tctgatgcaa tatggacaat tggtttcttc tctgaatggc gggagtatga
     5041 aaagtatggc tgaagcgcaa aatgatcccc tgctgccggg atactcgttt aatgcccatc
     5101 tggtggcggg tttaacgccg attgaggcca acggttatct cgattttttt atcgaccgac
     5161 cgctgggaat gaaaggttat attctcaatc tcaccattcg cggtcagggg gtggtgaaaa
     5221 atcagggacg agaatttgtt tgccgaccgg gtgatatttt gctgttcccg ccaggagaga
     5281 ttcatcacta cggtcgtcat ccggaggctc gcgaatggta tcaccagtgg gtttactttc
     5341 gtccgcgcgc ctactggcat gaatggctta actggccgtc aatatttgcc aatacggggt
     5401 tctttcgccc ggatgaagcg caccagccgc atttcagcga cctgtttggg caaatcatta
     5461 acgccgggca aggggaaggg cgctattcgg agctgctggc gataaatctg cttgagcaat
     5521 tgttactgcg gcgcatggaa gcgattaacg agtcgctcca tccaccgatg gataatcggg
     5581 tacgcgaggc ttgtcagtac atcagcgatc acctggcaga cagcaatttt gatatcgcca
     5641 gcgtcgcaca gcatgtttgc ttgtcgccgt cgcgtctgtc acatcttttc cgccagcagt
     5701 tagggattag cgtcttaagc tggcgcgagg accaacgtat cagccaggcg aagctgcttt
     5761 tgagcaccac ccggatgcct atcgccaccg tcggtcgcaa tgttggtttt gacgatcaac
     5821 tctatttctc gcgggtattt aaaaaatgca ccggggccag cccgagcgag ttccgtgccg
     5881 gttgtgaaga aaaagtgaat gatgtagccg tcaagttgtc ataaactagt ctgcagacgt
     5941 aaaaaaagcg gcgtggttag ccgctttttt aattgccgga gcgcaataaa aaagcccccg
     6001 gaaggtgatc ttccgggggc tttctcatgc gttgcttgta ttgaaaatgg gggccatgcc
     6061 tacctatctg cctcccgggt ccttgagccc cattatcacc tccaaagcct tctcgtctgg
     6121 tcagtttcac ctgttttacg taaaaacccg cttcggcggg tttttacttt tggggaaaca
     6181 cagaaaaaag cccgcacctg acagtgcggg cttttttttt cgaccaaagg tgcgactact
     6241 cttgcctact acctatcgac tgagctgaaa gaattccggt tctggcaaat attctgaaat
     6301 gagctgttga caattaatca tccggctcgt ataattctag tcgcgcggtt ctcgccaagg
     6361 ggtttaagag ctatgctgga aacagcatag caagtttaaa taaggctagt ccgttatcaa
     6421 cttgaaaaag tggcaccgag tcggtgcttt ttttttgaat tcatgtggct gaccgttctg
     6481 ttgtctctcg ctcttccgag tagacgaaca ataaggcctc cctaacgggg ggcctttttt
     6541 attgataaca aaagtcagtg cttccgctat ttccaaaata ccgggctaat acggtttaaa
     6601 cgacctcctg gatttgctca gacagccttt tcgtcattcg tttcagccaa aaaacttaag
     6661 accgccggtc ttgtccacta ccttgcagta atgcggtgga caggatcggc ggttttcttt
     6721 tctcttctca agaagttcct atactttcta gagaatagga acttcggaat aggaacttcc
     6781 tcctgaacgg ccataagaac gaaggctgtc tgttgaactc tcgagccttg tcgccttgcg
     6841 tataatattt gcccatggac gcacaccgtg gaaacggatg aaggcacgaa cccagttgac
     6901 ataagcctgt tcggttcgta aactgtaatg caagtagcgt atgcgctcac gcaactggtc
     6961 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     7021 tgactgtttt tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     7081 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagcaac gatgttacgc
     7141 agcagggcag tcgccctaaa acaaagttag gtggctcaag tatgggcatc attcgcacat
     7201 gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg
     7261 agttcggaga cgtagccacc tactcccaac atcagccgga ctccgattac ctcgggaact
     7321 tgctccgtag taagacattc atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg
     7381 ctctcgcggc ttacgttctg cccaagtttg agcagccgcg tagtgagatc tatatctatg
     7441 atctcgcagt ctccggagag caccggaggc agggcattgc caccgcgctc atcaatctcc
     7501 tcaagcatga ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg
     7561 acgatcccgc agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg
     7621 atatcgaccc aagtaccgcc acctaacaat tcgttcaagc cgagatctcg agcatcagct
     7681 tcaaaagcgc tctgaagttc ctatactttc tagagaatag gaacttcgga ataggtactt
     7741 caagatcccc aattcgagct catcgtccgg gccgcaagct cctagcggcg gatttgtcct
     7801 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
     7861 agcctttcgt tttatttgat gcctcaagct agagagtcat taccccaggc gtttaagggc
     7921 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tctagcttgg
     7981 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     8041 ttctgaggtc attactggat ctatcaacag gagtccaagc tcagctaatt aagctagctt
     8101 atcgataccg tcgacctcga accccacgcc cctctttaat acgacgggca atttgcactt
     8161 cagaaaatga agagtttgct ttagccataa caaaagtcca gtatgctttt tcacagcata
     8221 actggactga tttcagttta caactattct gtctagttta agactttatt gtcatagttt
     8281 agatctattt tgttcagttt aagactttat tgtccgccca cacctgctta cgcagggcat
     8341 ccatttatta ctcaaccgta accgattttg ccaggttacg cggctggtcc tctagctggc
     8401 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     8461 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     8521 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     8581 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     8641 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcgtca ccgaaacgcg
     8701 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     8761 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     8821 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     8881 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     8941 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     9001 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     9061 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     9121 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     9181 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     9241 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     9301 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     9361 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     9421 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     9481 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     9541 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     9601 ctggagccgg tgagcgtggg tcccgcggta tcattgcagc actggggcca gatggtaagc
     9661 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     9721 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     9781 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     9841 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     9901 cgtcagaccc cgtagaaaag atcaaaggca gttattggtg cctcactgat taagcattgg
     9961 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    10021 tttttgcggc cgcaagatcc ggccacgatg cgtccggcgt agaggatctg aagatcagca
    10081 gttcaacctg ttgatagtac gtactaagct ctcatgtttc acgtactaag ctctcatgtt
    10141 taacgtacta agctctcatg tttaacgaac taaaccctca tggctaacgt actaagctct
    10201 catggctaac gtactaagct ctcatgtttc acgtactaag ctctcatgtt tgaacaataa
    10261 aattaatata aatcagcaac ttaaatagcc tctaaggttt taagttttat aagaaaaaaa
    10321 agaatatata aggcttttaa agcttttaag gtttaacggt tgtggacaac aagccaggga
    10381 tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat tttgagtgac acaggaacac
    10441 ttaacggctg acatgggaat tagccatggc atcacagtat cgtgatgaca gaggcaggga
    10501 gtgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    10561 cgttatcagg ggattcctta aggtatactt tccgctgcat aaccctgctt cggggtcatt
    10621 atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg
    10681 gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta
    10741 ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc
    10801 aacgggaatc ctgctctgcg aggctggccg ataagct
//
