LOCUS       sequence-232703-       11230 bp    DNA     circular SYN 15-APR-2025
DEFINITION  control sgRNA in cells lacking rfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11230)
  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 11230)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pJMP1341
FEATURES             Location/Qualifiers
     source          1..11230
                     /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"
     protein_bind    4716..4732
                     /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        5114..5191
                     /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             5192..6274
                     /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(5211..5230)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6559..6575
                     /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)."
     protein_bind    complement(6978..7025)
                     /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(7102..7107,7108..7124,7125..7130)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             7241..8035
                     /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(7295..7314)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7905..7924
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(8087..8134)
                     /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(8185..8271)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(8374..8468)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(8496..8512)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8497..8513)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(8927..8946)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9046..9068
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9106..9124)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9192..9296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9297..9365,9366..10157)
                     /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(9515..9534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10458..10846
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            11115..11223
                     /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 ccatactagt
     4681 tttcctgtgt gaaacctgct gatgtgctca gtatcttgtt atccgctcac aatgtcaatg
     4741 ttatccgctc acatttataa tattttatct gattaataag atgatcttct tgagatcgtt
     4801 ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt
     4861 cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttggt gataagctgt
     4921 caaaccagat caattcgcac tagtctgcag acgtaaaaaa agcggcgtgg ttagccgctt
     4981 ttttaattgc cggagcgcaa taaaaaagcc cccggaaggt gatcttccgg gggctttctc
     5041 atgcgttgct tgtattgaaa atgggggcca tgcctaccta tctgcctccc gggaagcggc
     5101 atgcatttac gttgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
     5161 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     5221 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     5281 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     5341 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     5401 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     5461 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     5521 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     5581 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     5641 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     5701 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     5761 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     5821 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     5881 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     5941 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     6001 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     6061 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     6121 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     6181 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6241 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     6301 tcactcatta ggcaccccac ccgggtcctt gagccccatt atcacctcca aagccttctc
     6361 gtctggtcag tttcacctgt tttacgtaaa aacccgcttc ggcgggtttt tacttttggg
     6421 gaaacacaga aaaaagcccg cacctgacag tgcgggcttt ttttttcgac caaaggtgcg
     6481 actactcttg cctactacct atcgactgag ctgaaagaat tcctaaagat ctataaatgt
     6541 gagcggataa cattgacatt gtgagcggat aacaagatac ttctagtaac tttcagttta
     6601 gcggtctgtt taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt
     6661 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ttgaattcat gtggctgacc
     6721 gttctgttgt ctctcgctct tccgagtaga cgaacaataa ggcctcccta acggggggcc
     6781 ttttttattg ataacaaaag tcagtgcttc cgctatttcc aaaataccgg gctaatacgg
     6841 tttaaacgac ctcctggatt tgctcagaca gccttttcgt cattcgtttc agccaaaaaa
     6901 cttaagaccg ccggtcttgt ccactacctt gcagtaatgc ggtggacagg atcggcggtt
     6961 ttcttttctc ttctcaagaa gttcctatac tttctagaga ataggaactt cggaatagga
     7021 acttcctcct gaacggccat aagaacgaag gctgtctgtt gaactctcga gccgtggaaa
     7081 cggatgaagg cacgaaccca gttgacataa gcctgttcgg ttcgtaaact gtaatgcaag
     7141 tagcgtatgc gctcaaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     7201 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     7261 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     7321 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     7381 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     7441 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     7501 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     7561 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     7621 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     7681 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     7741 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     7801 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     7861 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     7921 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     7981 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     8041 actctggggt tcgaaatgac tcgagcatca gcttcaaaag cgctctgaag ttcctatact
     8101 ttctagagaa taggaacttc ggaataggta cttcaagatc cccaattcga gctcatcgtc
     8161 cgggccgcaa gctcctagcg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     8221 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctcaa
     8281 gctagagagt cattacccca ggcgtttaag ggcaccaata actgccttaa aaaaattacg
     8341 ccccgccctg ccactcatcg cagtctagct tggattctca ccaataaaaa acgcccggcg
     8401 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     8461 caggagtcca agctcagcta attaagctag cttatcgata ccgtcgacct cgaaccccac
     8521 gcccctcttt aatacgacgg gcaatttgca cttcagaaaa tgaagagttt gctttagcca
     8581 taacaaaagt ccagtatgct ttttcacagc ataactggac tgatttcagt ttacaactat
     8641 tctgtctagt ttaagacttt attgtcatag tttagatcta ttttgttcag tttaagactt
     8701 tattgtccgc ccacacctgc ttacgcaggg catccattta ttactcaacc gtaaccgatt
     8761 ttgccaggtt acgcggctgg tcctctagct ggcgtaatag cgaagaggcc cgcaccgatc
     8821 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     8881 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     8941 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     9001 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     9061 gtcagaggtt ttcaccgtcg tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     9121 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     9181 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     9241 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     9301 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     9361 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     9421 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     9481 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     9541 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     9601 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     9661 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     9721 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     9781 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     9841 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     9901 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     9961 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtcccgcg
    10021 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
    10081 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
    10141 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
    10201 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
    10261 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
    10321 gcagttattg gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
    10381 atactttaga ttgatttaaa acttcatttt taatttttgc ggccgcaaga tccggccacg
    10441 atgcgtccgg cgtagaggat ctgaagatca gcagttcaac ctgttgatag tacgtactaa
    10501 gctctcatgt ttcacgtact aagctctcat gtttaacgta ctaagctctc atgtttaacg
    10561 aactaaaccc tcatggctaa cgtactaagc tctcatggct aacgtactaa gctctcatgt
    10621 ttcacgtact aagctctcat gtttgaacaa taaaattaat ataaatcagc aacttaaata
    10681 gcctctaagg ttttaagttt tataagaaaa aaaagaatat ataaggcttt taaagctttt
    10741 aaggtttaac ggttgtggac aacaagccag ggatgtaacg cactgagaag cccttagagc
    10801 ctctcaaagc aattttgagt gacacaggaa cacttaacgg ctgacatggg aattagccat
    10861 ggcatcacag tatcgtgatg acagaggcag ggagtgcggc ctttttacgg ttcctggcct
    10921 tttgctggcc ttttgctcac atgttctttc ctgcgttatc aggggattcc ttaaggtata
    10981 ctttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc
    11041 tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc
    11101 caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc
    11161 ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg
    11221 ccgataagct
//
