LOCUS       sequence-476136-       14264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genome editing in Caulobacter crescentus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14264)
  AUTHORS   Sun J, Yu X, Tang G, Chen M, Zheng Y, Hu Y, Li Q, Li X, Li N, Li Z, 
            Li Y, Lu N, Tan W, Yang Y, Zeng X, Zhao G, Wang H, Dai L, Zhao G, Ai
            L, Zhao W
  TITLE     A CRISPR/SpCas9M-reporting system for efficient and rapid genome 
            editing in Caulobacter crescentus
  JOURNAL   bioRxiv 2024.12.02.626314
REFERENCE   2  (bases 1 to 14264)
  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: "bioRxiv 
            2024.12.02.626314"
COMMENT     Sequence Label: pSJX024
FEATURES             Location/Qualifiers
     source          1..14264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             458..1252
                     /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(512..531)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1122..1141
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        1580..1614
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        1635..1710
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      2596..2605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      5738..5784
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             6158..10261
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6158..10261
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     RBS             9718..9726
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      10271..10280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10277..10936
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(10343..10362)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     10955..10971
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     10956..10972
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11006..11022)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11006..11022)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(11054..11073)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(11055..11073)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11083..11100)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11083..11099)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11092..11114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(11808..12470)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAEHWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      12471..13242
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
      241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
      301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
      361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
      421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
     1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatata agctttcagt
     1561 cggcgcgaat gctccacgct tgacagctag ctcagtccta ggtataatac tagttctctt
     1621 gatgagatcg acgggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     1681 caacttgaaa aagtggcacc gagtcggtgc tttttttaca atcacccgat cgcccgcggc
     1741 gaaatccaga tcagcgacga tcgcatcgct atccaggtga cccgcgcggc tgacgtctac
     1801 gactacatgg ccggcggcag ctgatccgtc gcgggcttga gttcgacatt ctcgaacttc
     1861 cggcttccca aagccgattc agtttgctac cagccgcccc tcaccacgag cggtcgtggc
     1921 ggaattggta gacgcgcagc gttgaggtcg ctgtggggca acccgtggaa gttcgagtct
     1981 tctcgaccgc accaaggtga acaagagtag ggcgcgcttg gcgaaatcgg tttccgactc
     2041 gctgggggcg ctctaatcgt ttctggccct gacttcgcct cagggtcgct tgatcggacc
     2101 agcccaacga acccacgcgc tctgagagct caggaggccc ccatgacccg ggacaccgcc
     2161 gaactgaacg aagacgtgct ggacaacatc ccgctggccc atatcccgct tcccaagtcc
     2221 gagacgcccg aggagctgga agacctcgcc ctcggcgacg actacgcgct taatcccgaa
     2281 tatgtcgaga tggtgatcga cgccgccgac cgcggcgacg ccaagcgcct gcgcgagctg
     2341 gtgggggccc tggaccccgc cgacgtcgcc gacctgatgg gctttctgac ggccgactat
     2401 cgcgaggaag tgatcccgct gctcgacccc gaggcgctgg gcgagatcat ttccgagctg
     2461 gaggacaaca tccgcgagga agtgctggag gccaccccgt cggtgaccct ggcccgggcc
     2521 ctcgaggagc tggacaccga cgacgccgcc gacgttatcg acgacctgga cgacaccaag
     2581 cgcttccagg tgctggccgc catgggcgag accgaccgcg ccgccatcga gacgaccctg
     2641 tcctacgagg acgagaccgc cggccgcctg atgcagcgcg agttcctggc cgcgccggaa
     2701 ttctggaccg tcggccagac gatcgaccat gtccgcgagg ccggcgactc gctgcccgag
     2761 ctgttcttcg acgtctatgt cgtcgacccg accttcaagc ctgtcggggc gatcccgatc
     2821 agcatcctgc tgcgcagccg tcgtcagacg ccgctggccg agctgatgga ggcggtcacc
     2881 gaaatcccgg tcgagatgga ccaggaagag gtcgcctaca ttttcgacaa gtaccacctg
     2941 atcagcgccc cggtgattga accgggcggg cggctggtgg gtcagatcac tgtcgacgac
     3001 atcgtcgagg ttatccgcga cgaggccgag gaagacattc tggccctggc cggtgtgtcg
     3061 gacgccggcc gtgacgcctc ggtgatcgag atcgtcaagt cgcgcctgcc gtggctgatg
     3121 ctgaacctgg cgaccgccac cctggccgtc agcggcgtgg ccatcttcca ggccgagatc
     3181 gctcaattgg tggccctggc catcctgatg ccgatcgtct cgtccctggg cggcaacgcc
     3241 ggcacccaga ccctggccgt ggccgtgcgg gctctggcca gcaaggagct gacccccgcc
     3301 aacgcccgcc gcatcgtggc gcgggaactg ctagtgggcc tggttaacgg gctttgcctg
     3361 gcgctggtga tgggggcggc gacctacatg ttcttcgggc cgtcggatca acacgacaag
     3421 ctggcgatca tcgtcggcgc ggccctgatc accaacatct tcaccgcggc cctgggcggc
     3481 atcctgatgc ccctggcgct ggagaagatg ggccgtgacc cggcggtgtc gtcctcgatc
     3541 ttcgtcacct tcctgaccga cttcaccggc ttcttcgcgg tgctgctgat cgcctcgctg
     3601 ctgctgcact gaggttttct cgctggccac gaccctcgcc aggggaaact ggcgccgacc
     3661 ttgcgccggg acaggcgaac gcatccgcgt ctgtctcaaa ctgggtcaat ctaggcggtt
     3721 tcttgcctcg ccaccccgag gcatgagcaa gaacgcccgc tggtcccaaa gactggcggg
     3781 cgttttttgc tatcgtgcgg ctgttttctg gtggaaatcg accgagatcg gatgcacacg
     3841 ctctctcagt gttgagccag agcgtgacaa gcgctgaaac cgcccacatc ttccgctcgg
     3901 gcgttcggga aggaaaccat gtcgatgact ttcgcaccct cgatggactt cgcgctgggc
     3961 gagaccgctg acgcgatccg tgagaccacg gcccgtttcg ccgccgacaa gatcgcaccg
     4021 atcgccgcca agatcgacga gaccaacagc ttcccccgcg aactctgggt cccgatgggg
     4081 gatctgggtc tgcatggcat caccgtcgag gaagagttcg gggggctggg tctgggctat
     4141 ctcgaacacg tcgtggcgat ggaagaggtc agccgcgcct ccgcctccgt ggggctcagc
     4201 tacggcgccc actccaatct ctgcgtgaac cagatccgtc ggtgggccac gccggagcag
     4261 aaggcgcgct atctgcccaa gctgatcagc ggcgagcatg tgggctccct ggcgatgagc
     4321 gaagccggcg cgggctctga tgtcgtctcg atgaagctgc gcgccgagca ggtgggcgac
     4381 cgctacatcc tcaacggcac gaagttctgg atcaccaacg ccccgcacgc cgatacgctg
     4441 gtggtctacg ccaagaccgg cgagggcagc cgcggcatca ccgccttcat cgtcgagaag
     4501 ggcatgaagg gcttcagcgt ctccaagaag ctggacaaga tgggcatgcg cgggtcggac
     4561 accgccgagc tggtgttcga agactgcgag atccccgaag agaacgtcat gggcccggtc
     4621 ggcggtggcg tcggggtgct gatgagcggc ctcgactacg agcgggccgt gctggcggcc
     4681 ggtccgctgg gcatcatgca ggcttgcctc gatgtcgtgc tgccctacgt ccgcgaccgc
     4741 aagcagttcg gccagccgat cggctcgttc cagctgatgc agggcaagat cgccgacatg
     4801 tacgtggccc tgaactcggc ccgggcttac gtctacgcgg tggccaaggc ttgtgacgcg
     4861 ggcaagacca cgcgcttcga cgccgccggc gcgatcctga tggcgtcgga aaacgccgtg
     4921 aaggtgtcgc tggaggcgat ccaggccctc ggtggcgctg gctataccaa ggagtggccg
     4981 gtcgagcgcc tgctgcgcga cgccaagctc tatgacatcg gcgccggcac caacgagatc
     5041 cgccgcttcc tgatcggacg cgagctgatc ggcggctgat cctgggctgc gcgacatcaa
     5101 aagggccgcc ccgatcgctc ggggcggccc tccatcgtcg tagccgctct cgcagccagt
     5161 gactcgaaac ccttggcgct caacgcctta ggcggagacc tcgctcttcg ggctccaggt
     5221 cacgacttgc gaccgcggtt ggagacaatg agacactgga ccacctcctt tcagttgttg
     5281 aacaacacag gaaggatatt ggaccggttt tcgcgaaagg aaagggtcgt tatcgcgacc
     5341 aagcgggact aaacgccgcg cttcgctcgg ccaggctggg ccgcttgcga agatcggtga
     5401 tggggcgatg taccccgtcg agggcgaagc cggcggcgac gtagccggcc ttcacggcgg
     5461 gttcgaaggc gcgccagtca cggatatcga tgccttccag cttcggggtg atcagaacgt
     5521 cggtcgcctc gcgagcggcg gccagatcgc gcccggtcgt gacggtcgca gcacgcatca
     5581 gcaaggccac gatcggcggg cccttgcgcc attcgccgga gagaatccag ttccacaggg
     5641 atgggggcgt gacgatgtca gcgctggtga tgctgcggcc acgggtgacg tcgaccccga
     5701 cgatcggtcc aagctggcgc gcccagcggc gcggatccaa ataaaacgaa aggctcagtc
     5761 gaaagactgg gcctttcgtt ttatctgttg tttgtcgctg cagccagccg tggtcgggca
     5821 gcaggtagaa ggcgccctcg tcctgatcct cgcccgaaac ctccagcccc cggtcgatgg
     5881 cttcgacgac atagccggcc gcgcggcagg tgtcggtgag cgcggccagc agggcggctt
     5941 cctggtcagg ggtcaggtcg gtcatgggca agaggtccag gtcgtggttt gtcggcggct
     6001 tctagcatgg accgcccgcg cccgtgaggc cgaggatttc gcgctggtca gacaacctac
     6061 ttgccgtccc cacatgttag cgctaccaag tgccgacgaa cgcgcgccgc cgacggtgtc
     6121 ggcgcttcag acgctcgagt tttggggaga cgaccatatg gacaagaagt actcgatcgg
     6181 cctggatatc gggacgaaca gcgtcgggtg ggccgtgatc acggacgaat acaaggtgcc
     6241 ctccaagaag ttcaaggtcc tggggaatac cgaccggcac tccatcaaga agaacctcat
     6301 cggggccctg ctctttgact ccggggaaac ggccgaagcg acgcggctga agcggaccgc
     6361 gcgtcgtcgc tacacgcgcc ggaagaatcg catctgttat ctccaggaaa tcttcagcaa
     6421 cgagatggcc aaggtggatg attccttctt ccaccgcctg gaggagtcct ttctggtcga
     6481 agaggataag aagcatgagc gccacccgat ctttgggaac atcgtcgacg aagtggccta
     6541 ccacgaaaag tatcccacga tctaccacct gcgtaagaag ctcgtggact ccaccgataa
     6601 ggcggatctg cggctgatct acctcgccct ggcccacatg atcaagtttc ggggccattt
     6661 tctgatcgaa ggcgatctga atccggacaa tagcgacgtc gataagctct ttatccagct
     6721 ggtgcagacc tacaatcaac tcttcgagga gaatccgatc aatgcctccg gggtggatgc
     6781 caaggcgatc ctgagcgcgc gtctgtcgaa gtcccgccgc ctggagaacc tgatcgccca
     6841 gctcccgggc gaaaagaaga acgggctctt tgggaatctg atcgccctca gcctggggct
     6901 gaccccgaac tttaagtcca atttcgacct cgcggaagat gccaagctcc aactctcgaa
     6961 ggatacgtac gacgatgacc tcgacaacct gctggcccaa atcggcgacc agtacgcgga
     7021 cctgttcctg gcggccaaga acctgtcgga cgccatcctc ctgagcgaca tcctccgtgt
     7081 caatacggaa atcacgaagg cgccgctgtc cgccagcatg atcaagcggt atgatgagca
     7141 tcatcaagac ctcaccctgc tgaaggcgct ggtccggcaa cagctgcccg agaagtacaa
     7201 ggagatcttt ttcgatcaaa gcaagaacgg ctacgccggg tatatcgacg ggggggcctc
     7261 gcaagaagag ttctacaagt ttatcaagcc catcctggag aagatggatg ggacggagga
     7321 gctcctcgtc aagctcaacc gggaagatct cctccgcaag caacgcacct ttgataatgg
     7381 gtccatcccg catcaaatcc acctcggcga gctccatgcg atcctgcggc ggcaagagga
     7441 cttctacccg tttctcaagg acaaccgcga gaagatcgag aagatcctga cgttccgcat
     7501 cccctattat gtcggccccc tggcccgggg gaattcgcgg tttgcctgga tgacccgcaa
     7561 gtccgaagaa acgatcaccc cctggaattt tgaagaggtg gtggacaagg gggcgagcgc
     7621 gcagtccttt atcgaacgga tgaccaattt cgacaagaac ctccccaacg agaaggtgct
     7681 cccgaagcat tccctcctgt atgagtactt taccgtctac aacgagctca cgaaggtcaa
     7741 gtatgtcacc gagggcatgc gcaagcccgc cttcctctcg ggggaacaaa agaaggccat
     7801 cgtggacctg ctcttcaaga ccaaccgcaa ggtcaccgtg aagcaactca aggaggacta
     7861 tttcaagaag atcgaatgct tcgatagcgt ggagatctcc ggggtcgaag accggtttaa
     7921 cgcctcgctc ggcacgtacc atgatctcct caagatcatc aaggacaagg acttcctcga
     7981 caacgaggag aacgaagaca tcctggaaga catcgtgctg accctgaccc tcttcgagga
     8041 tcgcgagatg atcgaggagc gtctcaagac gtacgcgcat ctcttcgatg ataaggtgat
     8101 gaagcaactc aagcggcggc gttacacggg ctgggggcgc ctctcccgta agctcatcaa
     8161 cggcatccgg gacaagcaat cggggaagac catcctggac ttcctgaagt cggacggctt
     8221 tgccaatcgc aattttatgc aactcatcca cgacgattcc ctcaccttca aggaagatat
     8281 ccagaaggcc caggtgtccg ggcaagggga ctccctgcat gaacatatcg ccaacctggc
     8341 cgggagcccc gcgatcaaga aggggatcct ccagaccgtg aaggtggtcg atgagctcgt
     8401 gaaggtcatg ggccggcata agcccgaaaa tatcgtcatc gagatggcgc gcgagaacca
     8461 gaccacccaa aagggccaga agaattcgcg ggagcgcatg aagcggatcg aagaaggcat
     8521 caaggaactc ggcagccaga tcctgaagga acatccggtc gaaaacacgc agctccagaa
     8581 cgagaagctc tacctctact atctccaaaa tggccgggat atgtacgtgg accaggagct
     8641 cgacatcaat cggctcagcg actatgatgt ggatcatatc gtcccccaat cctttctcaa
     8701 ggacgacagc atcgataaca aggtcctgac ccggtcggac aagaatcgtg gcaagagcga
     8761 taacgtcccc tccgaggagg tcgtgaagaa gatgaagaac tattggcgcc aactgctcaa
     8821 cgccaagctg atcacgcaac ggaagttcga caacctcacc aaggccgagc gcgggggcct
     8881 ctcggaactg gacaaggcgg ggttcatcaa gcgccagctc gtcgaaaccc gtcaaatcac
     8941 caagcacgtg gcgcaaatcc tggatagccg catgaacacc aagtacgatg agaacgacaa
     9001 gctgatccgt gaagtgaagg tgatcaccct gaagtccaag ctcgtgagcg atttccgcaa
     9061 ggatttccaa ttttataagg tgcgggaaat caataattac catcacgcgc atgatgcgta
     9121 tctcaacgcc gtggtcggca cggccctgat caagaagtat cccaagctgg agtcggaatt
     9181 cgtctacggc gattacaagg tgtatgacgt ccgtaagatg atcgcgaaga gcgagcaaga
     9241 aatcgggaag gccacggcga agtacttctt ctacagcaat atcatgaatt tcttcaagac
     9301 ggagatcacg ctggcgaatg gcgaaatccg taagcgcccg ctgatcgaaa cgaatggcga
     9361 gaccggcgaa atcgtgtggg ataaggggcg cgatttcgcg acggtccgca aggtcctctc
     9421 gatgccgcag gtcaacatcg tcaagaagac cgaagtccag accggggggt tttccaagga
     9481 gagcatcctg ccgaagcgca acagcgacaa gctcatcgcc cgcaagaagg actgggatcc
     9541 caagaagtat ggcgggttcg atagcccgac cgtggcgtac tcggtgctgg tggtcgcgaa
     9601 ggtcgaaaag gggaagtcca agaagctcaa gagcgtcaag gaactgctgg ggatcacgat
     9661 catggaacgc tcctcctttg agaagaatcc catcgacttc ctggaggcga agggctataa
     9721 ggaggtgaag aaggatctca tcatcaagct gccgaagtat tcgctcttcg aactcgaaaa
     9781 tgggcgcaag cgcatgctgg cgtccgccgg cgagctccaa aagggcaatg agctggccct
     9841 gccctccaag tacgtcaatt ttctgtacct ggcctcccac tatgagaagc tgaagggcag
     9901 ccccgaggat aatgaacaga agcaactctt tgtcgagcaa cataagcatt acctggatga
     9961 aatcatcgag caaatcagcg aattttccaa gcgggtgatc ctcgccgatg ccaacctgga
    10021 caaggtcctc agcgcgtata acaagcaccg ggacaagccc atccgtgaac aagcggagaa
    10081 catcatccat ctgttcacgc tcaccaacct gggcgccccc gccgcgttta agtattttga
    10141 taccaccatc gatcgcaagc gttatacgtc caccaaggaa gtcctcgatg ccacgctgat
    10201 ccatcaaagc atcaccgggc tctacgagac ccggatcgat ctctcccagc tgggcggcga
    10261 tcaccggtcg gccaccatgg agaaaaaaat cactggatat accaccgttg atatatccca
    10321 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
    10381 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
    10441 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat
    10501 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
    10561 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
    10621 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
    10681 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
    10741 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
    10801 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
    10861 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
    10921 gcagggcggg gcgtaaatga aaacgggccc cccctcgagg tcgacggtat cgataagctt
    10981 gatatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg
    11041 gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
    11101 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
    11161 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
    11221 cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
    11281 ttttgttaaa tcagctcatt ttttaaccaa taggccgtac tgcgatgagt ggcagggcgg
    11341 ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggttgct acgcctgaat
    11401 aagtgataat aagcggatga atggcagaaa ttcgaaagca aattcgaccc ggtcgtcggt
    11461 tcagggcagg gtcgttaaat agccgcttat gtctattgct ggtttaccgg tttattgact
    11521 accggaagca gtgtgaccgt gtgcttctca aatgcctgag gccagtttgc tcaggctctc
    11581 cccgtggagg taataattga cgatatgatc atttattctg cctcccagag cctgataaaa
    11641 acggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc
    11701 atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgagg cggctacagc
    11761 cgatagtctg gaacagcgca cttacgggtt gctgcgcaac ccaagtgcta ccggcgcggc
    11821 agcgtgaccc gtgtcggcgg ctccaacggc tcgccatcgt ccagaaaaca cggctcatcg
    11881 ggcatcggca ggcgctgctg cccgcgccgt tcccattcct ccgtttcggt caaggctggc
    11941 aggtctggtt ccatgcccgg aatgccgggc tggctgggcg gctcctcgcc ggggccggtc
    12001 ggtagttgct gctcgcccgg atacagggtc gggatgcggc gcaggtcgcc atgccccaac
    12061 agcgattcgt cctggtcgtc gtgatcaacc accacggcgg cactgaacac cgacaggcgc
    12121 aactggtcgc ggggctggcc ccacgccacg cggtcattga ccacgtaggc cgacacggtg
    12181 ccggggccgt tgagcttcac gacggagatc cagtgctcgg ccaccaagtc cttgactgcg
    12241 tattggaccg tccgcaaaga acgtccgatg agcttggaaa gtgtcttctg gctgaccacc
    12301 acggcgttct ggtggcccat ctgcgccacg aggtgatgca gcagcattgc cgccgtgggt
    12361 ttcctcgcaa taagcccggc ccacgcctca tgcgctttgc gttccgtttg cacccagtga
    12421 ccgggcttgt tcttggcttg aatgccgatt tctctggact gcgtggccat gcttatctcc
    12481 atgcggtagg ggtgccgcac ggttgcggca ccatgcgcaa tcagctgcaa cttttcggca
    12541 gcgcgacaac aattatgcgt tgcgtaaaag tggcagtcaa ttacagattt tctttaacct
    12601 acgcaatgag ctattgcggg gggtgccgca atgagctgtt gcgtaccccc cttttttaag
    12661 ttgttgattt ttaagtcttt cgcatttcgc cctatatcta gttctttggt gcccaaagaa
    12721 gggcacccct gcggggttcc cccacgcctt cggcgcggct ccccctccgg caaaaagtgg
    12781 cccctccggg gcttgttgat cgactgcgcg gccttcggcc ttgcccaagg tggcgctgcc
    12841 cccttggaac ccccgcactc gccgccgtga ggctcggggg gcaggcgggc gggcttcgcc
    12901 cttcgactgc ccccactcgc ataggcttgg gtcgttccag gcgcgtcaag gccaagccgc
    12961 tgcgcggtcg ctgcgcgagc cttgacccgc cttccacttg gtgtccaacc ggcaagcgaa
    13021 gcgcgcaggc cgcaggccgg aggcttttcc ccagagaaaa ttaaaaaaat tgatggggca
    13081 aggccgcagg ccgcgcagtt ggagccggtg ggtatgtggt cgaaggctgg gtagccggtg
    13141 ggcaatccct gtggtcaagc tcgtgggcag gcgcagcctg tccatcagct tgtccagcag
    13201 ggttgtccac gggccgagcg aagcgagcca gccggtggcc gctcgcggcc atcgtccaca
    13261 tatccacggg ctggcaaggg agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc
    13321 cgtagggcgc cgcagccgcc gtaggcggtc acgactttgc gaagcaaagt ctagtgagta
    13381 tactcaagca ttgagtggcc cgccggaggc accgccttgc gctgcccccg tcgagccggt
    13441 tggacaccaa aagggagggg caggcatggc ggcatacgcg atcatgcgat gcaagaagct
    13501 ggcgaaaatg ggcaacgtgg cggccagtct caagcacgcc taccgcgagc gcgagacgcc
    13561 caacgctgac gccagcagga cgccagagaa cgagcactgg gcggccagca gcaccgatga
    13621 agcgatgggc cgactgcgcg agttgctgcc agagaagcgg cgcaaggacg ctgtgttggc
    13681 ggtcgagtac gtcatgacgg ccagcccgga atggtggaag tcggccagcc aagaacagca
    13741 ggcggcgttc ttcgagaagg cgcacaagtg gctggcggac aagtacgggg cggatcgcat
    13801 cgtgacggcc agcatccacc gtgacgaaac cagcccgcac atgaccgcgt tcgtggtgcc
    13861 gctgacgcag gacggcaggc tgtcggccaa ggagttcatc ggcaacaaag cgcagatgac
    13921 ccgcgaccag accacgtttg cggccgctgt ggccgatcta gggctgcaac ggggcatcga
    13981 gggcagcaag gcacgtcaca cgcgcattca ggcgttctac gaggccctgg agcggccacc
    14041 agtgggccac gtcaccatca gcccgcaagc ggtcgagcca cgcgcctatg caccgcaggg
    14101 attggccgaa aagctgggaa tctcaaagcg cgttgagacg ccggaagccg tggccgaccg
    14161 gctgacaaaa gcggttcggc aggggtatga gcctgcccta caggccgccg caggagcgcg
    14221 tgagatgcgc aagaaggccg atcaagccca agagacggcc cgag
//
