LOCUS       sequence-237226-       16600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Cas9 nuclease and RecAb recombination system
            in Acinetobacter baumannii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16600)
  AUTHORS   Wang Y, Wang Z, Chen Y, Hua X, Yu Y, Ji Q
  TITLE     A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in 
            Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of 
            OxyR.
  JOURNAL   Cell Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 
            10.1016/j.chembiol.2019.09.003.
  PUBMED    31548010
REFERENCE   2  (bases 1 to 16600)
  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: "Cell Chem 
            Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 
            10.1016/j.chembiol.2019.09.003."
COMMENT     Sequence Label: pCasAb-apr
FEATURES             Location/Qualifiers
     source          1..16600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..55)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      208..294
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      386..413
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(678..683,684..699,700..706)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    714..730
                     /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             753..4859
                     /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             753..4856
                     /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             join(5271..5357,5358..6692)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     primer_bind     complement(6731..6750)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6754..6770)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6755..6771)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             6909..7709
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      8149..8543
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            8885..8972
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             10211..11182
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             11693..12532
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             12519..13370
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(13647..14729)
                     /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     14691..14710
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(14730..14807)
                     /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."
     promoter        join(15037..15042,15043..15058,15059..15065)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    15073..15089
                     /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)."
ORIGIN
        1 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
       61 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
      121 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
      181 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      241 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
      301 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
      361 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
      421 ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      481 ataaccctgg atccgggctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag
      541 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
      601 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
      661 tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
      721 cggataacaa tttcacacag gaaacagaat tcatggataa gaaatactca ataggcttag
      781 atatcggcac aaatagcgtc ggatgggcgg tgatcactga tgaatataag gttccgtcta
      841 aaaagttcaa ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg
      901 ctcttttatt tgacagtgga gagacagcgg aagcgactcg tctcaaacgg acagctcgta
      961 gaaggtatac acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga
     1021 tggcgaaagt agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag
     1081 acaagaagca tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg
     1141 agaaatatcc aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg
     1201 atttgcgctt aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga
     1261 ttgagggaga tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac
     1321 aaacctacaa tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag
     1381 cgattctttc tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc
     1441 ccggtgagaa gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc
     1501 ctaattttaa atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata
     1561 cttacgatga tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt
     1621 ttttggcagc taagaattta tcagatgcta ttttactttc agatatccta agagtaaata
     1681 ctgaaataac taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc
     1741 aagacttgac tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa
     1801 tcttttttga tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag
     1861 aagaatttta taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat
     1921 tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta
     1981 ttccccatca aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt
     2041 atccattttt aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt
     2101 attatgttgg tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg
     2161 aagaaacaat taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat
     2221 catttattga acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa
     2281 aacatagttt gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg
     2341 ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg
     2401 atttactctt caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca
     2461 aaaaaataga atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt
     2521 cattaggtac ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg
     2581 aagaaaatga agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg
     2641 agatgattga ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac
     2701 agcttaaacg tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta
     2761 ttagggataa gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca
     2821 atcgcaattt tatgcagctg atccatgatg atagtttgac atttaaagaa gacattcaaa
     2881 aagcacaagt gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta
     2941 gccctgctat taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag
     3001 taatggggcg gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa
     3061 ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag
     3121 aattaggaag tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa
     3181 agctctatct ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata
     3241 ttaatcgttt aagtgattat gatgtcgatc acattgttcc acaaagtttc cttaaagacg
     3301 attcaataga caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg
     3361 ttccaagtga agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca
     3421 agttaatcac tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg
     3481 aacttgataa agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc
     3541 atgtggcaca aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta
     3601 ttcgagaggt taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt
     3661 tccaattcta taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa
     3721 atgccgtcgt tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct
     3781 atggtgatta taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag
     3841 gcaaagcaac cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa
     3901 ttacacttgc aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg
     3961 gagaaattgt ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc
     4021 cccaagtcaa tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa
     4081 ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa
     4141 aatatggtgg ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg
     4201 aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg
     4261 aaagaagttc ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag
     4321 ttaaaaaaga cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc
     4381 gtaaacggat gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa
     4441 gcaaatatgt gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag
     4501 aagataacga acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta
     4561 ttgagcaaat cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag
     4621 ttcttagtgc atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta
     4681 ttcatttatt tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa
     4741 caattgatcg taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc
     4801 aatccatcac tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgactgat
     4861 ggctggttgg cgtactgttg tggtaaatac caagcttgca actttatgcc catgcaacag
     4921 aaactataaa aaatacagag aatgaaaaga aacagataga ttttttagtt ctttaggccc
     4981 gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata gaccagttgc
     5041 aatccaaacg agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg tttgttactg
     5101 ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg tatactttgg cgtcacccct
     5161 tacatatttt aggtcttttt ttattgtgcg taactaactt gccatcttca aacaggaggg
     5221 ctggaagaag cagaccgcta acacagtaca taaaaaagga gacatgaacg atgaacatca
     5281 aaaagtttgc aaaacaagca acagtattaa cctttactac cgcactgctg gcaggaggcg
     5341 caactcaagc gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca tacggcattt
     5401 cccatattac acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat gaaaaatatc
     5461 aagttcctga gttcgattcg tccacaatta aaaatatctc ttctgcaaaa ggcctggacg
     5521 tttgggacag ctggccatta caaaacgctg acggcactgt cgcaaactat cacggctacc
     5581 acatcgtctt tgcattagcc ggagatccta aaaatgcgga tgacacatcg atttacatgt
     5641 tctatcaaaa agtcggcgaa acttctattg acagctggaa aaacgctggc cgcgtcttta
     5701 aagacagcga caaattcgat gcaaatgatt ctatcctaaa agaccaaaca caagaatggt
     5761 caggttcagc cacatttaca tctgacggaa aaatccgttt attctacact gatttctccg
     5821 gtaaacatta cggcaaacaa acactgacaa ctgcacaagt taacgtatca gcatcagaca
     5881 gctctttgaa catcaacggt gtagaggatt ataaatcaat ctttgacggt gacggaaaaa
     5941 cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta cagctcaggc gacaaccata
     6001 cgctgagaga tcctcactac gtagaagata aaggccacaa atacttagta tttgaagcaa
     6061 acactggaac tgaagatggc taccaaggcg aagaatcttt atttaacaaa gcatactatg
     6121 gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc gataaaaaac
     6181 gcacggctga gttagcaaac ggcgctctcg gtatgattga gctaaacgat gattacacac
     6241 tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt aacagatgaa attgaacgcg
     6301 cgaacgtctt taaaatgaac ggcaaatggt atctgttcac tgactcccgc ggatcaaaaa
     6361 tgacgattga cggcattacg tctaacgata tttacatgct tggttatgtt tctaattctt
     6421 taactggccc atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg gatcttgatc
     6481 ctaacgatgt aacctttact tactcacact tcgctgtacc tcaagcgaaa ggaaacaatg
     6541 tcgtgattac aagctatatg acaaacagag gattctacgc agacaaacaa tcaacgtttg
     6601 cgcctagctt cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa gacagcatcc
     6661 ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa aagaaaatgc cgattatggt
     6721 gcactctcag tacaatctgc tctgatgccg tatcgatacc gtcgacctcg agggggggcc
     6781 cggtaccatc ccaatacgcg tcaattcacg gatccggttc atgtgcagct ccatcagcaa
     6841 aaggggatga taagtttatc accaccgact atttgcaaca gtgccgttga tcgtgctatg
     6901 atcgactgat gtcatcagcg gtggagtgca atgtcgtgca atacgaatgg cgaaaagccg
     6961 agctcatcgg tcagcttctc aaccttgggg ttacccccgg cggtgtgctg ctggtccaca
     7021 gctccttccg tagcgtccgg cccctcgaag atgggccact tggactgatc gaggccctgc
     7081 gtgctgcgct gggtccggga gggacgctcg tcatgccctc gtggtcaggt ctggacgacg
     7141 agccgttcga tcctgccacg tcgcccgtta caccggacct tggagttgtc tctgacacat
     7201 tctggcgcct gccaaatgta aagcgcagcg cccatccatt tgcctttgcg gcagcggggc
     7261 cacaggcaga gcagatcatc tctgatccat tgcccctgcc acctcactcg cctgcaagcc
     7321 cggtcgcccg tgtccatgaa ctcgatgggc aggtacttct cctcggcgtg ggacacgatg
     7381 ccaacacgac gctgcatctt gccgagttga tggcaaaggt tccctatggg gtgccgagac
     7441 actgcaccat tcttcaggat ggcaagttgg tacgcgtcga ttatctcgag aatgaccact
     7501 gctgtgagcg ctttgccttg gcggacaggt ggctcaagga gaagagcctt cagaaggaag
     7561 gtccagtcgg tcatgccttt gctcggttga tccgctcccg cgacattgtg gcgacagccc
     7621 tgggtcaact gggccgagat ccgttgatct tcctgcatcc gccagaggcg ggatgcgaag
     7681 aatgcgatgc cgctcgccag tcgattggct gactgtcaga ccaagtttac tcatatatac
     7741 tttagattga tttctgaaag cgaccaggtg ctcggcgtgg caagactcgc agcgaacccg
     7801 tagaaagcca tgctccagcc gcccgcattg gagaaattct tcaaattccc gttgcacata
     7861 gcccggcaat tcctttccct gctctgccat aagcgcagcg aatgccgggt aatactcgtc
     7921 aacgatctga tagagaaggg tttgctcggg tcggtggctc tggtaacgac cagtatcccg
     7981 atcccggctg gccgtcctgg ccgccacatg aggcatgttc cgcgtccttg caatactgtg
     8041 tttacataca gtctatcgct tagcggaaag ttcttttacc ctcagccgaa atgcctgccg
     8101 ttgctagaca ttgccagcca gtgcccgtca ctcccgtact aactgtcacg aacccctgca
     8161 ataactgtca cgcccccctg caataactgt cacgaacccc tgcaataact gtcacgcccc
     8221 caaacctgca aacccagcag gggcgggggc tggcggggtg ttggaaaaat ccatccatga
     8281 ttatctaaga ataatccact aggcgcggtt atcagcgccc ttgtggggcg ctgctgccct
     8341 tgcccaatat gcccggccag aggccggata gctggtctat tcgctgcgct aggctacaca
     8401 ccgccccacc gctgcgcggc agggggaaag gcgggcaaag cccgctaaac cccacaccaa
     8461 accccgcaga aatacgctgg agcgctttta gccgctttag cggcctttcc ccctacccga
     8521 agggtggggg cgcgtgtgca gccccgcagg gcctgtctcg gtcgatcatt cagcccggct
     8581 catccttctg gcgtggcggc agaccgaaca aggcgcggtc gtggtcgcgt tcaaggtacg
     8641 catccattgc cgccatgagc cgatcctccg gccactcgct gctgttcacc ttggccaaaa
     8701 tcatggcccc caccagcacc ttgcgccttg tttcgttctt gcgctcttgc tgctgttccc
     8761 ttgcccgcac ccgctgaatt tcggcattga ttcgcgctcg ttgttcttcg agcttggcca
     8821 gccgatccgc cgccttgttg ctccccttaa ccatcttgac accccattgt taatgtgctg
     8881 tctcgtaggc tatcatggag gcacagcggc ggcaatcccg accctacttt gtaggggagg
     8941 gcgcacttac cggtttctct tcgagaaact ggcctaacgg ccacccttcg ggcggtgcgc
     9001 tctccgaggg ccattgcatg gagccgaaaa gcaaaagcaa cagcgaggca gcatggcgat
     9061 ttatcacctt acggcgaaaa ccggcagcag gtcgggcggc caatcggcca gggccaaggc
     9121 cgactacatc cagcgcgaag gcaagtatgc ccgcgacatg gatgaagtct tgcacgccga
     9181 atccgggcac atgccggagt tcgtcgagcg gcccgccgac tactgggatg ctgccgacct
     9241 gtatgaacgc gccaatgggc ggctgttcaa ggaggtcgaa tttgccctgc cggtcgagct
     9301 gaccctcgac cagcagaagg cgctggcgtc cgagttcgcc cagcacctga ccggtgccga
     9361 gcgcctgccg tatacgctgg ccatccatgc cggtggcggc gagaacccgc actgccacct
     9421 gatgatctcc gagcggatca atgacggcat cgagcggccc gccgctcagt ggttcaagcg
     9481 gtacaacggc aagaccccgg agaagggcgg ggcacagaag accgaagcgc tcaagcccaa
     9541 ggcatggctt gagcagaccc gcgaggcatg ggccgaccat gccaaccggg cattagagcg
     9601 ggctggccac gacgcccgca ttgaccacag aacacttgag gcgcagggca tcgagcgcct
     9661 gcccggtgtt cacctggggc cgaacgtggt ggagatggaa ggccggggca tccgcaccga
     9721 ccgggcagac gtggccctga acatcgacac cgccaacgcc cagatcatcg acttacagga
     9781 ataccgggag gcaatagacc atgaacgcaa tcgacagagt gaagaaatcc agaggcatca
     9841 acgagttagc ggagcagatc gaaccgctgg cccagagcat ggcgacactg gccgacgaag
     9901 cccggcaggt catgagccag acccagcagg ccagcgaggc gcaggcggcg gagtggctga
     9961 aagcccagcg ccagacaggg gcggcatggg tggagctggc caaagagttg cgggaggtag
    10021 ccgccgaggt gagcagcgcc gcgcagagcg cccggagcgc gtcgcggggg tggcactgga
    10081 agctatggct aaccgtgatg ctggcttcca tgatgcctac ggtggtgctg ctgatcgcat
    10141 cgttgctctt gctcgacctg acgccactga caaccgagga cggctcgatc tggctgcgct
    10201 tggtggcccg atgaagaacg acaggacttt gcaggccata ggccgacagc tcaaggccat
    10261 gggctgtgag cgcttcgata tcggcgtcag ggacgccacc accggccaga tgatgaaccg
    10321 ggaatggtca gccgccgaag tgctccagaa cacgccatgg ctcaagcgga tgaatgccca
    10381 gggcaatgac gtgtatatca ggcccgccga gcaggagcgg catggtctgg tgctggtgga
    10441 cgacctcagc gagtttgacc tggatgacat gaaagccgag ggccgggagc ctgccctggt
    10501 agtggaaacc agcccgaaga actatcaggc atgggtcaag gtggccgacg ccgcaggcgg
    10561 tgaacttcgg gggcagattg cccggacgct ggccagcgag tacgacgccg acccggccag
    10621 cgccgacagc cgccactatg gccgcttggc gggcttcacc aaccgcaagg acaagcacac
    10681 cacccgcgcc ggttatcagc cgtgggtgct gctgcgtgaa tccaagggca agaccgccac
    10741 cgctggcccg gcgctggtgc agcaggctgg ccagcagatc gagcaggccc agcggcagca
    10801 ggagaaggcc cgcaggctgg ccagcctcga actgcccgag cggcagctta gccgccaccg
    10861 gcgcacggcg ctggacgagt accgcagcga gatggccggg ctggtcaagc gcttcggtga
    10921 tgacctcagc aagtgcgact ttatcgccgc gcagaagctg gccagccggg gccgcagtgc
    10981 cgaggaaatc ggcaaggcca tggccgaggc cagcccagcg ctggcagagc gcaagcccgg
    11041 ccacgaagcg gattacatcg agcgcaccgt cagcaaggtc atgggtctgc ccagcgtcca
    11101 gcttgcgcgg gccgagctgg cacgggcacc ggcaccccgc cagcgaggca tggacagggg
    11161 cgggccagat ttcagcatgt agtgcttgcg ttggtactca cgcctgttat actatgagta
    11221 ctcacgcaca gaagggggtt ttatggaata cgaaaaaagc gcttcagggt cggtctacct
    11281 gatcaaaagt gacaagggct attggttgcc cggtggcttt ggttatacgt caaacaaggc
    11341 cgaggctggc cgcttttcag tcgctgatat ggccagcctt aaccttgacg gctgcacctt
    11401 gtccttgttc cgcgaagaca agcctttcgg ccccggcaag tttctcggtg actgatatga
    11461 aagaccaaaa ggacaagcag accggcgacc tgctggccag ccctgacgct gtacgccaag
    11521 cgcgatatgc cgagcgcatg aaggccaaag ggatgcgtca gcgcaagttc tggctgaccg
    11581 acgacgaata cgaggcgctg cgcgagtgcc tggaagaact cagagcggcg cagggcgggg
    11641 gtagtgaccc cgccagcgcc taaccaccaa ctgcctgcaa aggaggcaat caatggctac
    11701 ccataagcct atcaatattc tggaggcgtt cgcagcagcg ccgccaccgc tggactacgt
    11761 tttgcccaac atggtggccg gtacggtcgg ggcgctggtg tcgcccggtg gtgccggtaa
    11821 atccatgctg gccctgcaac tggccgcaca gattgcaggc gggccggatc tgctggaggt
    11881 gggcgaactg cccaccggcc cggtgatcta cctgcccgcc gaagacccgc ccaccgccat
    11941 tcatcaccgc ctgcacgccc ttggggcgca cctcagcgcc gaggaacggc aagccgtggc
    12001 tgacggcctg ctgatccagc cgctgatcgg cagcctgccc aacatcatgg ccccggagtg
    12061 gttcgacggc ctcaagcgcg ccgccgaggg ccgccgcctg atggtgctgg acacgctgcg
    12121 ccggttccac atcgaggaag aaaacgccag cggccccatg gcccaggtca tcggtcgcat
    12181 ggaggccatc gccgccgata ccgggtgctc tatcgtgttc ctgcaccatg ccagcaaggg
    12241 cgcggccatg atgggcgcag gcgaccagca gcaggccagc cggggcagct cggtactggt
    12301 cgataacatc cgctggcagt cctacctgtc gagcatgacc agcgccgagg ccgaggaatg
    12361 gggtgtggac gacgaccagc gccggttctt cgtccgcttc ggtgtgagca aggccaacta
    12421 tggcgcaccg ttcgctgatc ggtggttcag gcggcatgac ggcggggtgc tcaagcccgc
    12481 cgtgctggag aggcagcgca agagcaaggg ggtgccccgt ggtgaagcct aagaacaagc
    12541 acagcctcag ccacgtccgg cacgacccgg cgcactgtct ggcccccggc ctgttccgtg
    12601 ccctcaagcg gggcgagcgc aagcgcagca agctggacgt gacgtatgac tacggcgacg
    12661 gcaagcggat cgagttcagc ggcccggagc cgctgggcgc tgatgatctg cgcatcctgc
    12721 aagggctggt ggccatggct gggcctaatg gcctagtgct tggcccggaa cccaagaccg
    12781 aaggcggacg gcagctccgg ctgttcctgg aacccaagtg ggaggccgtc accgctgatg
    12841 ccatggtggt caaaggtagc tatcgggcgc tggcaaagga aatcggggca gaggtcgata
    12901 gtggtggggc gctcaagcac atacaggact gcatcgagcg cctttggaag gtatccatca
    12961 tcgcccagaa tggccgcaag cggcaggggt ttcggctgct gtcggagtac gccagcgacg
    13021 aggcggacgg gcgcctgtac gtggccctga accccttgat cgcgcaggcc gtcatgggtg
    13081 gcggccagca tgtgcgcatc agcatggacg aggtgcgggc gctggacagc gaaaccgccc
    13141 gcctgctgca ccagcggctg tgtggctgga tcgaccccgg caaaaccggc aaggcttcca
    13201 tagatacctt gtgcggctat gtctggccgt cagaggccag tggttcgacc atgcgcaagc
    13261 gccgccagcg ggtgcgcgag gcgttgccgg agctggtcgc gctgggctgg acggtaaccg
    13321 agttcgcggc gggcaagtac gacatcaccc ggcccaaggc ggcaggctga ccccccccac
    13381 tctattgtaa acaagacatt tttatctttt atattcaatg gcttattttc ctgctaattg
    13441 gtaataccat gaaaaatacc atgctcagaa aaggcttaac aatattttga aaaattgcct
    13501 actgagcgct gccgcacagc tccataggcc gctttcctgg ctttgcttcc agatgtatgc
    13561 tcttctgctc ccgaacgcca gcaagacgta gcccagcgcg tcggccagct tgcaattcgc
    13621 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    13681 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
    13741 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
    13801 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
    13861 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
    13921 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
    13981 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
    14041 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
    14101 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
    14161 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
    14221 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
    14281 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
    14341 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
    14401 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
    14461 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
    14521 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
    14581 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
    14641 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
    14701 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
    14761 cgctatcatg ccataccgcg aaaggttttg caccattcga tggtgtcaac gtaaatgccg
    14821 cttcgccttc gcgcgcgaat tgcaagctga tccgggctta tcgactgcac ggtgcaccaa
    14881 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
    14941 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
    15001 tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta
    15061 taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagaatt cgagctcggt
    15121 accaggagga aacgatgaat gcgccagtaa atggaacact tattactaca cagattgcaa
    15181 acgttgctga aactcttggc ttggttaatg ttaatccaca agagttaaag gaaacactga
    15241 ttcaaacagc tttccgtact gaaacacctg caactgatgc acaaatggct tctcttttga
    15301 ttgttgctgg tcaatacaag ctgaacccat ggaccaaaga gatttacgct ttcccagata
    15361 aaaacaaagg gattattcca gttgttggcg tggatggttg gtctcgaatc attaatggaa
    15421 actctaattt caatggtatg gaatttaagt tttctgaaaa tatggttcag atggaaggcg
    15481 cgaaagttgc tgcacctgaa tgggttgaat gcattatcta tcgtaaagat cgtgaccacc
    15541 ctactgttgt tcgcgagtat ttagcagagt gttatcgtgc accatttaag tcaaaaactg
    15601 gatatgtagt tgaaggacca tggcagagtc acccttctcg cttcttgcgc cacaaagcaa
    15661 ctattcagtg tgcacgctta gcctttggtt ttgtgggtat tcatgatcag gatgaagcag
    15721 aacgtatcgc tgaaagtgga caagctatta aggatgtgac tagtgaagtg ccagaaggct
    15781 accaagcctt tgaagatgag catttgccta cactcaaatc agaagctcaa tacggcactg
    15841 aacgcttgca agctgcttat gtggcaattc caaagggaaa tcttaaaaag cacctttggg
    15901 aagttcactc aattagctta aaaaaaattg ctcagtttgc agaccaagct ttacagcgcc
    15961 aaggagaaac atatgaacat tctccagcgt agtgaagatt ggcattcgga acgctgtggg
    16021 aaagtcacag caagccgtgt aaaggattta aatgcaaagc tcaataaagg aaaagcttta
    16081 aatgcattgg gtttaactat tctagctgag cgcctcactg gcgttcagaa ggaaatcttc
    16141 actaaccaag ctatgcaatg gggtatcgat aacgagcctc atgcaatagc agcttatgaa
    16201 aatgaaacgg gtaactttgt agtaggtaca ggtttaattg accatccttt cattgaaatg
    16261 ttcggggctt caccagatgg gcttgtgggc gacaatgggc aaatagaagt taagtgccca
    16321 gatactacaa cacacttgaa caccctgctg accaagcaag taccagatga gtacattcct
    16381 caaataacta gtcagttggc ttgtactcgt cgtgaatggt gtgactttgt gagttatgac
    16441 ccacgtctac cagaaggatt acagatcatc attattcgtg tctatgccaa agacctagca
    16501 atagaagcac ttgagcaaga tgttcgtaag ttcaacaaag ctatagatga cgcaattaaa
    16561 acattgaagg tggcagcatg agtcgacctg caggcatgca
//
