LOCUS       sequence-478586-       14015 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 14015)
  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 14015)
  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: pSJX022
FEATURES             Location/Qualifiers
     source          1..14015
                     /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             5924..10027
                     /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             5924..10027
                     /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             9484..9492
                     /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)"
     CDS             10028..10687
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(10094..10113)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     10706..10722
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     10707..10723
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10757..10773)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10757..10773)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(10805..10824)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10806..10824)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10834..10851)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10834..10850)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10843..10865)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(11559..12221)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      12222..12993
                     /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 tttgtcgggc tggactctag ccgaccgact
     5821 gagacgctca caagcgttca attggatcca atcttgacgt ccgtttgatt acgatcaaga
     5881 ttggatccaa taggtcgccg aaccacgatg cgaggaaacg catatggaca agaagtactc
     5941 gatcggcctg gatatcggga cgaacagcgt cgggtgggcc gtgatcacgg acgaatacaa
     6001 ggtgccctcc aagaagttca aggtcctggg gaataccgac cggcactcca tcaagaagaa
     6061 cctcatcggg gccctgctct ttgactccgg ggaaacggcc gaagcgacgc ggctgaagcg
     6121 gaccgcgcgt cgtcgctaca cgcgccggaa gaatcgcatc tgttatctcc aggaaatctt
     6181 cagcaacgag atggccaagg tggatgattc cttcttccac cgcctggagg agtcctttct
     6241 ggtcgaagag gataagaagc atgagcgcca cccgatcttt gggaacatcg tcgacgaagt
     6301 ggcctaccac gaaaagtatc ccacgatcta ccacctgcgt aagaagctcg tggactccac
     6361 cgataaggcg gatctgcggc tgatctacct cgccctggcc cacatgatca agtttcgggg
     6421 ccattttctg atcgaaggcg atctgaatcc ggacaatagc gacgtcgata agctctttat
     6481 ccagctggtg cagacctaca atcaactctt cgaggagaat ccgatcaatg cctccggggt
     6541 ggatgccaag gcgatcctga gcgcgcgtct gtcgaagtcc cgccgcctgg agaacctgat
     6601 cgcccagctc ccgggcgaaa agaagaacgg gctctttggg aatctgatcg ccctcagcct
     6661 ggggctgacc ccgaacttta agtccaattt cgacctcgcg gaagatgcca agctccaact
     6721 ctcgaaggat acgtacgacg atgacctcga caacctgctg gcccaaatcg gcgaccagta
     6781 cgcggacctg ttcctggcgg ccaagaacct gtcggacgcc atcctcctga gcgacatcct
     6841 ccgtgtcaat acggaaatca cgaaggcgcc gctgtccgcc agcatgatca agcggtatga
     6901 tgagcatcat caagacctca ccctgctgaa ggcgctggtc cggcaacagc tgcccgagaa
     6961 gtacaaggag atctttttcg atcaaagcaa gaacggctac gccgggtata tcgacggggg
     7021 ggcctcgcaa gaagagttct acaagtttat caagcccatc ctggagaaga tggatgggac
     7081 ggaggagctc ctcgtcaagc tcaaccggga agatctcctc cgcaagcaac gcacctttga
     7141 taatgggtcc atcccgcatc aaatccacct cggcgagctc catgcgatcc tgcggcggca
     7201 agaggacttc tacccgtttc tcaaggacaa ccgcgagaag atcgagaaga tcctgacgtt
     7261 ccgcatcccc tattatgtcg gccccctggc ccgggggaat tcgcggtttg cctggatgac
     7321 ccgcaagtcc gaagaaacga tcaccccctg gaattttgaa gaggtggtgg acaagggggc
     7381 gagcgcgcag tcctttatcg aacggatgac caatttcgac aagaacctcc ccaacgagaa
     7441 ggtgctcccg aagcattccc tcctgtatga gtactttacc gtctacaacg agctcacgaa
     7501 ggtcaagtat gtcaccgagg gcatgcgcaa gcccgccttc ctctcggggg aacaaaagaa
     7561 ggccatcgtg gacctgctct tcaagaccaa ccgcaaggtc accgtgaagc aactcaagga
     7621 ggactatttc aagaagatcg aatgcttcga tagcgtggag atctccgggg tcgaagaccg
     7681 gtttaacgcc tcgctcggca cgtaccatga tctcctcaag atcatcaagg acaaggactt
     7741 cctcgacaac gaggagaacg aagacatcct ggaagacatc gtgctgaccc tgaccctctt
     7801 cgaggatcgc gagatgatcg aggagcgtct caagacgtac gcgcatctct tcgatgataa
     7861 ggtgatgaag caactcaagc ggcggcgtta cacgggctgg gggcgcctct cccgtaagct
     7921 catcaacggc atccgggaca agcaatcggg gaagaccatc ctggacttcc tgaagtcgga
     7981 cggctttgcc aatcgcaatt ttatgcaact catccacgac gattccctca ccttcaagga
     8041 agatatccag aaggcccagg tgtccgggca aggggactcc ctgcatgaac atatcgccaa
     8101 cctggccggg agccccgcga tcaagaaggg gatcctccag accgtgaagg tggtcgatga
     8161 gctcgtgaag gtcatgggcc ggcataagcc cgaaaatatc gtcatcgaga tggcgcgcga
     8221 gaaccagacc acccaaaagg gccagaagaa ttcgcgggag cgcatgaagc ggatcgaaga
     8281 aggcatcaag gaactcggca gccagatcct gaaggaacat ccggtcgaaa acacgcagct
     8341 ccagaacgag aagctctacc tctactatct ccaaaatggc cgggatatgt acgtggacca
     8401 ggagctcgac atcaatcggc tcagcgacta tgatgtggat catatcgtcc cccaatcctt
     8461 tctcaaggac gacagcatcg ataacaaggt cctgacccgg tcggacaaga atcgtggcaa
     8521 gagcgataac gtcccctccg aggaggtcgt gaagaagatg aagaactatt ggcgccaact
     8581 gctcaacgcc aagctgatca cgcaacggaa gttcgacaac ctcaccaagg ccgagcgcgg
     8641 gggcctctcg gaactggaca aggcggggtt catcaagcgc cagctcgtcg aaacccgtca
     8701 aatcaccaag cacgtggcgc aaatcctgga tagccgcatg aacaccaagt acgatgagaa
     8761 cgacaagctg atccgtgaag tgaaggtgat caccctgaag tccaagctcg tgagcgattt
     8821 ccgcaaggat ttccaatttt ataaggtgcg ggaaatcaat aattaccatc acgcgcatga
     8881 tgcgtatctc aacgccgtgg tcggcacggc cctgatcaag aagtatccca agctggagtc
     8941 ggaattcgtc tacggcgatt acaaggtgta tgacgtccgt aagatgatcg cgaagagcga
     9001 gcaagaaatc gggaaggcca cggcgaagta cttcttctac agcaatatca tgaatttctt
     9061 caagacggag atcacgctgg cgaatggcga aatccgtaag cgcccgctga tcgaaacgaa
     9121 tggcgagacc ggcgaaatcg tgtgggataa ggggcgcgat ttcgcgacgg tccgcaaggt
     9181 cctctcgatg ccgcaggtca acatcgtcaa gaagaccgaa gtccagaccg gggggttttc
     9241 caaggagagc atcctgccga agcgcaacag cgacaagctc atcgcccgca agaaggactg
     9301 ggatcccaag aagtatggcg ggttcgatag cccgaccgtg gcgtactcgg tgctggtggt
     9361 cgcgaaggtc gaaaagggga agtccaagaa gctcaagagc gtcaaggaac tgctggggat
     9421 cacgatcatg gaacgctcct cctttgagaa gaatcccatc gacttcctgg aggcgaaggg
     9481 ctataaggag gtgaagaagg atctcatcat caagctgccg aagtattcgc tcttcgaact
     9541 cgaaaatggg cgcaagcgca tgctggcgtc cgccggcgag ctccaaaagg gcaatgagct
     9601 ggccctgccc tccaagtacg tcaattttct gtacctagcc tcccactatg agaagctgaa
     9661 gggcagcccc gaggataatg aacagaagca actctttgtc gagcaacata agcattacct
     9721 ggatgaaatc atcgagcaaa tcagcgaatt ttccaagcgg gtgatcctcg ccgatgccaa
     9781 cctggacaag gtcctcagcg cgtataacaa gcaccgggac aagcccatcc gtgaacaagc
     9841 ggagaacatc atccatctgt tcacgctcac caacctgggc gcccccgccg cgtttaagta
     9901 ttttgatacc accatcgatc gcaagcgtta tacgtccacc aaggaagtcc tcgatgccac
     9961 gctgatccat caaagcatca ccgggctcta cgagacccgg atcgatctct cccagctggg
    10021 cggcgatatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
    10081 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
    10141 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
    10201 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa
    10261 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
    10321 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
    10381 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
    10441 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
    10501 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
    10561 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
    10621 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
    10681 ggcgtaaatg aaaacgggcc ccccctcgag gtcgacggta tcgataagct tgatatcgaa
    10741 ttcctgcagc ccgggggatc cactagttct agagcggccg ccaccgcggt ggagctccaa
    10801 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
    10861 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
    10921 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
    10981 tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
    11041 atcagctcat tttttaacca ataggccgta ctgcgatgag tggcagggcg gggcgtaatt
    11101 tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa taagtgataa
    11161 taagcggatg aatggcagaa attcgaaagc aaattcgacc cggtcgtcgg ttcagggcag
    11221 ggtcgttaaa tagccgctta tgtctattgc tggtttaccg gtttattgac taccggaagc
    11281 agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct ccccgtggag
    11341 gtaataattg acgatatgat catttattct gcctcccaga gcctgataaa aacggtgaat
    11401 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
    11461 gacgcaacgc ggggaggcag acaaggtata gggcggcgag gcggctacag ccgatagtct
    11521 ggaacagcgc acttacgggt tgctgcgcaa cccaagtgct accggcgcgg cagcgtgacc
    11581 cgtgtcggcg gctccaacgg ctcgccatcg tccagaaaac acggctcatc gggcatcggc
    11641 aggcgctgct gcccgcgccg ttcccattcc tccgtttcgg tcaaggctgg caggtctggt
    11701 tccatgcccg gaatgccggg ctggctgggc ggctcctcgc cggggccggt cggtagttgc
    11761 tgctcgcccg gatacagggt cgggatgcgg cgcaggtcgc catgccccaa cagcgattcg
    11821 tcctggtcgt cgtgatcaac caccacggcg gcactgaaca ccgacaggcg caactggtcg
    11881 cggggctggc cccacgccac gcggtcattg accacgtagg ccgacacggt gccggggccg
    11941 ttgagcttca cgacggagat ccagcgctcg gccaccaagt ccttgactgc gtattggacc
    12001 gtccgcaaag aacgtccgat gagcttggaa agtgtcttct ggctgaccac cacggcgttc
    12061 tggtggccca tctgcgccac gaggtgatgc agcagcattg ccgccgtggg tttcctcgca
    12121 ataagcccgg cccacgcctc atgcgctttg cgttccgttt gcacccagtg accgggcttg
    12181 ttcttggctt gaatgccgat ttctctggac tgcgtggcca tgcttatctc catgcggtag
    12241 gggtgccgca cggttgcggc accatgcgca atcagctgca acttttcggc agcgcgacaa
    12301 caattatgcg ttgcgtaaaa gtggcagtca attacagatt ttctttaacc tacgcaatga
    12361 gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc ccttttttaa gttgttgatt
    12421 tttaagtctt tcgcatttcg ccctatatct agttctttgg tgcccaaaga agggcacccc
    12481 tgcggggttc ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg
    12541 ggcttgttga tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc ccccttggaa
    12601 cccccgcact cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc ccttcgactg
    12661 cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc
    12721 gctgcgcgag ccttgacccg ccttccactt ggtgtccaac cggcaagcga agcgcgcagg
    12781 ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa ttgatggggc aaggccgcag
    12841 gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg ggtagccggt gggcaatccc
    12901 tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc ttgtccagca gggttgtcca
    12961 cgggccgagc gaagcgagcc agccggtggc cgctcgcggc catcgtccac atatccacgg
    13021 gctggcaagg gagcgcagcg accgcgcagg gcgaagcccg gagagcaagc ccgtagggcg
    13081 ccgcagccgc cgtaggcggt cacgactttg cgaagcaaag tctagtgagt atactcaagc
    13141 attgagtggc ccgccggagg caccgccttg cgctgccccc gtcgagccgg ttggacacca
    13201 aaagggaggg gcaggcatgg cggcatacgc gatcatgcga tgcaagaagc tggcgaaaat
    13261 gggcaacgtg gcggccagtc tcaagcacgc ctaccgcgag cgcgagacgc ccaacgctga
    13321 cgccagcagg acgccagaga acgagcactg ggcggccagc agcaccgatg aagcgatggg
    13381 ccgactgcgc gagttgctgc cagagaagcg gcgcaaggac gctgtgttgg cggtcgagta
    13441 cgtcatgacg gccagcccgg aatggtggaa gtcggccagc caagaacagc aggcggcgtt
    13501 cttcgagaag gcgcacaagt ggctggcgga caagtacggg gcggatcgca tcgtgacggc
    13561 cagcatccac cgtgacgaaa ccagcccgca catgaccgcg ttcgtggtgc cgctgacgca
    13621 ggacggcagg ctgtcggcca aggagttcat cggcaacaaa gcgcagatga cccgcgacca
    13681 gaccacgttt gcggccgctg tggccgatct agggctgcaa cggggcatcg agggcagcaa
    13741 ggcacgtcac acgcgcattc aggcgttcta cgaggccctg gagcggccac cagtgggcca
    13801 cgtcaccatc agcccgcaag cggtcgagcc acgcgcctat gcaccgcagg gattggccga
    13861 aaagctggga atctcaaagc gcgttgagac gccggaagcc gtggccgacc ggctgacaaa
    13921 agcggttcgg caggggtatg agcctgccct acaggccgcc gcaggagcgc gtgagatgcg
    13981 caagaaggcc gatcaagccc aagagacggc ccgag
//
