LOCUS       sequence-232557-       11861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rfp ""test"" strain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11861)
  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 11861)
  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: pJMP1221
FEATURES             Location/Qualifiers
     source          1..11861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            8..116
                     /note="incP origin of transfer"
     rep_origin      385..773
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(1074..1865,1866..1934))
                     /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     1697..1716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1935..2039)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2107..2125
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2163..2185)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2285..2304
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2718..2734
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2719..2735
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2763..2857
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2960..3046
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    3097..3144
                     /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)."
     primer_bind     3808..3827
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(3976..3981,3982..3998,3999..4004))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    4081..4128
                     /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)."
     CDS             complement(4266..4943)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             4950..4961
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    5298..5314
                     /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)."
     CDS             complement(5596..6678)
                     /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     6640..6659
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6679..6756)
                     /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."
     protein_bind    7129..7145
                     /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)."
     CDS             join(7187..7213,7214..7216,7217..9703,9704..9706,
                     9707..11290)
                     /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             7187..11290
                     /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"
     CDS             11300..11389
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     mobile_element  11545..11710
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 agcttatcgg ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag
       61 ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct
      121 gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc
      181 gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg
      241 cagcggaaag tataccttaa ggaatcccct gataacgcag gaaagaacat gtgagcaaaa
      301 ggccagcaaa aggccaggaa ccgtaaaaag gccgcactcc ctgcctctgt catcacgata
      361 ctgtgatgcc atggctaatt cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt
      421 gctttgagag gctctaaggg cttctcagtg cgttacatcc ctggcttgtt gtccacaacc
      481 gttaaacctt aaaagcttta aaagccttat atattctttt ttttcttata aaacttaaaa
      541 ccttagaggc tatttaagtt gctgatttat attaatttta ttgttcaaac atgagagctt
      601 agtacgtgaa acatgagagc ttagtacgtt agccatgaga gcttagtacg ttagccatga
      661 gggtttagtt cgttaaacat gagagcttag tacgttaaac atgagagctt agtacgtgaa
      721 acatgagagc ttagtacgta ctatcaacag gttgaactgc tgatcttcag atcctctacg
      781 ccggacgcat cgtggccgga tcttgcggcc gcaaaaatta aaaatgaagt tttaaatcaa
      841 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      901 caataactgc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
      961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1201 gcaatgatac cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2041 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2101 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga cgacggtgaa
     2161 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2221 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     2281 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     2341 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     2401 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctagagga ccagccgcgt
     2461 aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcaggtgtgg
     2521 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa
     2581 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg
     2641 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt
     2701 aaagaggggc gtggggttcg aggtcgacgg tatcgataag ctagcttaat tagctgagct
     2761 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     2821 gctcggttgc cgccgggcgt tttttattgg tgagaatcca agctagactg cgatgagtgg
     2881 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
     2941 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     3001 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgctaggagc
     3061 ttgcggcccg gacgatgagc tcgaattggg gatcttgaag tacctattcc gaagttccta
     3121 ttctctagaa agtataggaa cttcagagcg cttttgaagc tgatgctcga gttccagtcg
     3181 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3241 cgtattgggc gcatgcataa aaactgttgt aattcattaa gcattctgcc gacatggaag
     3301 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     3361 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     3421 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     3481 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     3541 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     3601 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     3661 ttcattgcca tacggaactc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     3721 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     3781 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     3841 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     3901 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tgagcgcata
     3961 cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt tcgtgccttc
     4021 atccgtttcc acggctcgag agttcaacag acagccttcg ttcttatggc cgttcaggag
     4081 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttctt gagaagagaa
     4141 aagaaaaccg ccgatcctgt ccaccgcatt actgcaaggt agtggacaag accggcggtc
     4201 ttaagttttt tggctgaaac gaatgacgaa aaggctgtct gagcaaatcc aggaggtcgt
     4261 ttttattaag caccggtgga gtgacgacct tcagcacgtt cgtactgttc aacgatggtg
     4321 tagtcttcgt tgtgggaggt gatgtccagt ttgatgtcgg ttttgtaagc acccggcagc
     4381 tgaaccggtt ttttagccat gtaggtggtt ttaacttcag cgtcgtagtg accaccgtct
     4441 ttcagtttca gacgcatttt gatttcacct ttcagagcac cgtcttccgg gtacatacgt
     4501 tcggtggaag cttcccaacc catggttttt ttctgcataa ccggaccgtc ggacgggaag
     4561 ttggtaccac gcagtttaac tttgtagatg aactcaccgt cttgcaggga ggagtcctgg
     4621 gtaacggtaa caacaccacc gtcttcgaag ttcataacac gttcccattt gaaaccttcc
     4681 gggaaggaca gtttcaggta gtccgggatg tcagccgggt gtttaacgta agctttggaa
     4741 ccgtactgga actgcgggga caggatgtcc caagcgaacg gcagcggacc acctttggta
     4801 actttcagtt tagcggtctg ggtaccttcg tacggacgac cttcaccttc accttcgatt
     4861 tcgaactcgt gaccgttaac ggaaccttcc atacgaactt tgaaacgcat gaactctttg
     4921 ataacgtctt cgctactcgc catggtacct ttctcctctt taattaattc agatctatta
     4981 tacctaggac tgagctagct gtcaaattca ccaccctgaa ttgactctca aaccgtatta
     5041 gcccggtatt ttggaaatag cggaagcact gacttttgtt atcaataaaa aaggcccccc
     5101 gttagggagg ccttattgtt cgtctactcg gaagagcgag agacaacaga acggtcagcc
     5161 acatgaattc aaaaaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta
     5221 gccttattta aacttgctat gctgtttcca gcatagctct taaacagacc gctaaactga
     5281 aagttactag aagtatcttg ttatccgctc acaatgtcaa tgttatccgc tcacatttat
     5341 agatctttag gaattctttc agctcagtcg ataggtagta ggcaagagta gtcgcacctt
     5401 tggtcgaaaa aaaaagcccg cactgtcagg tgcgggcttt tttctgtgtt tccccaaaag
     5461 taaaaacccg ccgaagcggg tttttacgta aaacaggtga aactgaccag acgagaaggc
     5521 tttggaggtg ataatggggc tcaaggaccc tggggtgcct aatgagtgag ctaacttaca
     5581 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5641 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5701 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5761 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5821 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     5881 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5941 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     6001 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     6061 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     6121 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     6181 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     6241 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     6301 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     6361 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     6421 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     6481 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     6541 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     6601 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     6661 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     6721 cataccgcga aaggttttgc accattcgat ggtgtcaacg taaatgcatg ccgcttggga
     6781 ggcagatagg taggcatggc ccccattttc aatacaagca acgcatgaga aagcccccgg
     6841 aagatcacct tccgggggct tttttattgc gctccggcaa ttaaaaaagc ggctaaccac
     6901 gccgcttttt ttacgtctgc aggcgaattg atctggtttg acagcttatc accaagccag
     6961 ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg
     7021 gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat
     7081 cttattaatc agataaaata ttataaatgt gagcggataa cattgacatt gtgagcggat
     7141 aacaagatac tgagcacatc agcaggtttc acacaggaaa actagtatgg acaagaagta
     7201 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     7261 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     7321 gaacctgatc ggcgccctgc tgttcgacag cggagaaaca gccgaggcca cccggctgaa
     7381 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     7441 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     7501 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     7561 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     7621 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     7681 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     7741 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     7801 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     7861 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag
     7921 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     7981 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     8041 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     8101 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     8161 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     8221 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acatcgatgg
     8281 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     8341 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     8401 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     8461 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     8521 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     8581 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     8641 cgccagcgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     8701 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtaca acgagctgac
     8761 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     8821 aaaagccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     8881 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     8941 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     9001 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     9061 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     9121 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     9181 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     9241 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     9301 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     9361 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     9421 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     9481 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     9541 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     9601 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     9661 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gacgctatcg tgcctcagag
     9721 ctttctgaag gacgactcca tcgataacaa agtgctgact cggagcgaca agaaccgggg
     9781 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcgcca
     9841 gctgctgaat gccaagctga ttacccagag gaagttcgac aatctgacca aggccgagag
     9901 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     9961 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
    10021 gaacgacaaa ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
    10081 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
    10141 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
    10201 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
    10261 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
    10321 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
    10381 aaacggcgaa acaggcgaga tcgtgtggga taagggccgg gactttgcca ccgtgcggaa
    10441 agtgctgtct atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
    10501 cagcaaagag tctatcctgc ccaagaggaa cagcgacaag ctgatcgcca gaaagaagga
    10561 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
    10621 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
    10681 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
    10741 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
    10801 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
    10861 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
    10921 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaaacacta
    10981 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
    11041 taatctggac aaggtgctga gcgcctacaa caagcacaga gacaagccta tcagagagca
    11101 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
    11161 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
    11221 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
    11281 gggaggcgac gcggccgcgg agcagaaact catctctgaa gaagatctgg aacaaaagtt
    11341 gatttcagaa gaagatctgg aacagaagct catctctgag gaagatctgt aataaggcgc
    11401 gcctccttaa tgggacttgc agcctcggta ccaaattcca gaaaagacac ccgaaagggt
    11461 gttttttcgt tttggtccca cagaatgagc atcatggctc tagtcgacat cggagaagag
    11521 tacggctctt ttaaccgcct caagaaccag ataagtgaaa tctagttcca aactattttg
    11581 tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac
    11641 tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc aactattttg
    11701 tccgcccaca gcggggcatt tttcttcctg ttatgtttgg gcgctgcatt aatgaatcgg
    11761 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    11821 cccgctgcgc tcggtcgttc ggctgcggcg agcggtatca g
//
