LOCUS       sequence-228567-       11615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Cas9 nuclease and lambda-Red system in 
            Klebsiella Pneumoniae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11615)
  AUTHORS   Wang Y, Wang S, Chen W, Song L, Zhang Y, Shen Z, Yu F, Li M, Ji Q
  TITLE     Precise and efficient genome editing in Klebsiella pneumoniae using 
            CRISPR-Cas9 and CRISPR-assisted cytidine deaminase.
  JOURNAL   Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 
            10.1128/AEM.01834-18.
  PUBMED    30217854
REFERENCE   2  (bases 1 to 11615)
  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: "Appl 
            Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 
            10.1128/AEM.01834-18."
COMMENT     Sequence Label: pCasKP-hph
FEATURES             Location/Qualifiers
     source          1..11615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             344..760
                     /codon_start=1
                     /gene="gam"
                     /product="inhibitor of the host RecBCD nuclease in the 
                     lambda Red system"
                     /label=Gam
                     /note="protects the double-stranded DNA substrate for 
                     recombination"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             766..1551
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             1548..2228
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGEDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     CDS             2810..6916
                     /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             2810..6913
                     /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             complement(7051..8001)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      8049..8271
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        8901..9003
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(9070..9089)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(10711..11589)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagcgaaaa gatgtttcgt gaagccgtcg acgcttataa aaaatggata ttaatactga
      361 aactgagatc aagcaaaagc attcactaac cccctttcct gttttcctaa tcagcccggc
      421 atttcgcggg cgatattttc acagctattt caggagttca gccatgaacg cttattacat
      481 tcaggatcgt cttgaggctc agagctgggc gcgtcactac cagcagctcg cccgtgaaga
      541 gaaagaggca gaactggcag acgacatgga aaaaggcctg ccccagcacc tgtttgaatc
      601 gctatgcatc gatcatttgc aacgccacgg ggccagcaaa aaatccatta cccgtgcgtt
      661 tgatgacgat gttgagtttc aggagcgcat ggcagaacac atccggtaca tggttgaaac
      721 cattgctcac caccaggttg atattgattc agaggtataa aacgaatgag tactgcactc
      781 gcaacgctgg ctgggaagct ggctgaacgt gtcggcatgg attctgtcga cccacaggaa
      841 ctgatcacca ctcttcgcca gacggcattt aaaggtgatg ccagcgatgc gcagttcatc
      901 gcattactga tcgttgccaa ccagtacggc cttaatccgt ggacgaaaga aatttacgcc
      961 tttcctgata agcagaatgg catcgttccg gtggtgggcg ttgatggctg gtcccgcatc
     1021 atcaatgaaa accagcagtt tgatggcatg gactttgagc aggacaatga atcctgtaca
     1081 tgccggattt accgcaagga ccgtaatcat ccgatctgcg ttaccgaatg gatggatgaa
     1141 tgccgccgcg aaccattcaa aactcgcgaa ggcagagaaa tcacggggcc gtggcagtcg
     1201 catcccaaac ggatgttacg tcataaagcc atgattcagt gtgcccgtct ggccttcgga
     1261 tttgctggta tctatgacaa ggatgaagcc gagcgcattg tcgaaaatac tgcatacact
     1321 gcagaacgtc agccggaacg cgacatcact ccggttaacg atgaaaccat gcaggagatt
     1381 aacactctgc tgatcgccct ggataaaaca tgggatgacg acttattgcc gctctgttcc
     1441 cagatatttc gccgcgacat tcgtgcatcg tcagaactga cacaggccga agcagtaaaa
     1501 gctcttggat tcctgaaaca gaaagccgca gagcagaagg tggcagcatg acaccggaca
     1561 ttatcctgca gcgtaccggg atcgatgtga gagctgtcga acagggggaa gatgcgtggc
     1621 acaaattacg gctcggcgtc atcaccgctt cagaagttca caacgtgata gcaaaacccc
     1681 gctccggaaa gaagtggcct gacatgaaaa tgtcctactt ccacaccctg cttgctgagg
     1741 tttgcaccgg tgtggctccg gaagttaacg ctaaagcact ggcctgggga aaacagtacg
     1801 agaacgacgc cagaaccctg tttgaattca cttccggcgt gaatgttact gaatccccga
     1861 tcatctatcg cgacgaaagt atgcgtaccg cctgctctcc cgatggttta tgcagtgacg
     1921 gcaacggcct tgaactgaaa tgcccgttta cctcccggga tttcatgaag ttccggctcg
     1981 gtggtttcga ggccataaag tcagcttaca tggcccaggt gcagtacagc atgtgggtga
     2041 cgcgaaaaaa tgcctggtac tttgccaact atgacccgcg tatgaagcgt gaaggcctgc
     2101 attatgtcgt gattgagcgg gatgaaaagt acatggcgag ttttgacgag atcgtgccgg
     2161 agttcatcga aaaaatggac gaggcactgg ctgaaattgg ttttgtattt ggggagcaat
     2221 ggcgatgacg catcctcacg ataataagct tcctgctgaa catcaaaggc aagaaaacat
     2281 ctgttgtcaa agacagcatc cttgaacaag gacaattaac agttaacaaa taaaaacgca
     2341 aaagaaaatg ccgatattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg
     2401 attcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc
     2461 atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc
     2521 atctgttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa aaactcgtaa
     2581 aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatag ctatactgat
     2641 ttcgtcagac tcacagtcaa acatgccggt cagttggcct ggtgatggcg ggatcgttgt
     2701 atatttcttg acaccttttc ggcaccgccc taaaattctg cgtcctcata atatatgagg
     2761 cgatttatta cgtgtttacg aagcaaaagc taaaaccagg agctatttaa tggataagaa
     2821 atactcaata ggcttagata tcggcacaaa tagcgtcgga tgggcggtga tcactgatga
     2881 atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa
     2941 aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct
     3001 caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga
     3061 gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc
     3121 ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga
     3181 tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga
     3241 ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt
     3301 tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact
     3361 atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag
     3421 tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa
     3481 tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt
     3541 gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt
     3601 acagctttca aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga
     3661 tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt tactttcaga
     3721 tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa tgattaaacg
     3781 ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac aacaacttcc
     3841 agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag gttatattga
     3901 tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag aaaaaatgga
     3961 tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac
     4021 ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg ctattttgag
     4081 aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg aaaaaatctt
     4141 gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc gttttgcatg
     4201 gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag ttgtcgataa
     4261 aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa atcttccaaa
     4321 tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt ataacgaatt
     4381 gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca
     4441 gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt
     4501 aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt caggagttga
     4561 agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta ttaaagataa
     4621 agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt taacattgac
     4681 cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc acctctttga
     4741 tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac gtttgtctcg
     4801 aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag attttttgaa
     4861 atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata gtttgacatt
     4921 taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac atgaacatat
     4981 tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg taaaagttgt
     5041 tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta ttgaaatggc
     5101 acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat
     5161 cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg ttgaaaatac
     5221 tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag acatgtatgt
     5281 ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatcaca ttgttccaca
     5341 aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg ataaaaatcg
     5401 tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa actattggag
     5461 acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa cgaaagctga
     5521 acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat tggttgaaac
     5581 tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata ctaaatacga
     5641 tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta aattagtttc
     5701 tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt accatcatgc
     5761 ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat atccaaaact
     5821 tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa tgattgctaa
     5881 gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta atatcatgaa
     5941 cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga
     6001 aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg ccacagtgcg
     6061 caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac agacaggcgg
     6121 attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg ctcgtaaaaa
     6181 agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt attcagtcct
     6241 agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta aagagttact
     6301 agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact ttttagaagc
     6361 taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat atagtctttt
     6421 tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac aaaaaggaaa
     6481 tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc attatgaaaa
     6541 gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc agcataagca
     6601 ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta ttttagcaga
     6661 tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac caatacgtga
     6721 acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc ccgctgcttt
     6781 taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag aagttttaga
     6841 tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg atttgagtca
     6901 gctaggaggt gactgatggc tggttggcgt actgttgtgg taaataccca tatggacagt
     6961 tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct
     7021 tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt
     7081 ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt
     7141 actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa
     7201 agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt
     7261 tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat
     7321 ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac
     7381 caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg
     7441 gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc
     7501 aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca
     7561 ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt
     7621 tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa
     7681 tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa
     7741 aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc
     7801 ataagcattt tccctactga tgttcatcat ctgaacgtat tggttataag tgaacgatac
     7861 cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa
     7921 aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt
     7981 gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca
     8041 attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg
     8101 taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc
     8161 cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa
     8221 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     8281 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     8341 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     8401 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     8461 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     8521 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     8581 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     8641 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     8701 gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc ggggatcgac
     8761 gctctccctt atgcgactcc tgcacctttc gtcttcgaat aaatacctgt gacggaagat
     8821 cacttcgcag aataaataaa tcctggtgtc cctgttgata ccgggaagcc ctgggccaac
     8881 ttttggcgaa aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa
     8941 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct
     9001 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     9061 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     9121 ggtcatccac cggatcaatt cccctgctcg cgcaggctgg gtgccaagct ctcgggtaac
     9181 atcaaggccc gatccttgga gcccttgccc tcccgcacga tgatcgtgcc gtgatcgaaa
     9241 tccagatcct tgacccgcag ttgcaaaccc tcactgatcc ggctcacggt aactgatgcc
     9301 gtatttgcag taccagcgta cggcccacag aatgatgtca cgctgaaaat gccggccttt
     9361 gaatgggttc atgtgcagct ccatcagcaa aaggggatga taagtttatc accaccgact
     9421 atttgcaaca gtgccgttga tcgtgctatg atcgactgat gtcatcagcg gtggagtgca
     9481 atgtcatgac acaagaatcc ctgttacttc tcgaccgtat tgattcggat gattcctacg
     9541 cgagcctgcg gaacgaccag gaattctggg agccgctggc ccgccgagcc ctggaggagc
     9601 tcgggctgcc ggtgccgccg gtgctgcggg tgcccggcga gagcaccaac cccgtactgg
     9661 tcggcgagcc cggcccggtg atcaagctgt tcggcgagca ctggtgcggt ccggagagcc
     9721 tcgcgtcgga gtcggaggcg tacgcggtcc tggcggacgc cccggtgccg gtgccccgcc
     9781 tcctcgtccg cggcgagctg cggcccggca ccggagcctg gccgtggccc tacctggtga
     9841 tgagccggat gaccggcacc acctggcggt ccgcgatgga cggcacgacc gaccggaacg
     9901 cgctgctcgc cctggcccgc gaactcggcc gggtgctcgg ccggctgcac agggtgccgc
     9961 tgaccgggaa caccgtgctc accccccatt ccgaggtctt cccggaactg ctgcgggaac
    10021 gccgcgcggc gaccgtcgag gaccaccgcg ggtggggcta cctctcgccc cggctgctgg
    10081 accgcctgga ggactggctg ccggacgtgg acacgctgct ggccggccgc ggaccccggt
    10141 tcgtccacgg cgacctgcac gggaccaaca tcttcgtgga cctggccgcg accgaggtca
    10201 ccgggatcgt cgacttcacc gacgtctatg cgggagaccc ccgctacagt ctggtgcaac
    10261 tgcatctcaa cgccttccgg ggcgaccgcg agatcctggc cgcgctgctc gacggggcgc
    10321 agtggaagcg gaccgaggac ttcgcccgcg aactgctcgc cttcaccttc ctgcacgact
    10381 tcgaggtgtt cgaggagacc ccgctggatc tctccggctt caccgatccg gaggaactgg
    10441 cgcagttcct ctgggggccg ccggacaccg cccccggcgc ctgagctcat gagcggagaa
    10501 cgagatgacg ttggaggggc aaggtcgcgc tgattgctgg ggcaacacgt gaaaggcgag
    10561 atcaccaagg tagtcggcaa ataatgtcta acggtacccg gggatcctct agagtcgacc
    10621 tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt
    10681 caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac
    10741 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
    10801 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
    10861 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
    10921 gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac
    10981 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
    11041 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
    11101 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
    11161 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
    11221 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
    11281 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
    11341 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa
    11401 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
    11461 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
    11521 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
    11581 ttcagccata cttttcatac tcccgccatt cagag
//
