LOCUS       sequence-446029-       12652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombineering plasmid with constitutively expressed cas9 and the 
            araBAD promoter expressing alpha-beta-gamma.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12652)
  AUTHORS   Cook TB, Rand JM, Nurani W, Courtney DK, Liu SA, Pfleger BF
  TITLE     Genetic tools for reliable gene expression and recombineering in 
            Pseudomonas putida.
  JOURNAL   J Ind Microbiol Biotechnol. 2018 Jan 3. pii: 
            10.1007/s10295-017-2001-5. doi: 10.1007/s10295-017-2001-5.
  PUBMED    29299733
REFERENCE   2  (bases 1 to 12652)
  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: "J Ind 
            Microbiol Biotechnol. 2018 Jan 3. pii: 10.1007/s10295-017-2001-5. 
            doi: 10.1007/s10295-017-2001-5."
COMMENT     Sequence Label: pCas9
FEATURES             Location/Qualifiers
     source          1..12652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(62..1210)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALKNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      1603..2134
                     /label=oriV
                     /note="incP origin of replication"
     CDS             complement(2549..3082)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEVVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3252..3257,3258..3274,3275..3280))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             complement(3495..3518)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(3528..7631)
                     /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(3528..7631)
                     /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"
     promoter        complement(7667..7701)
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_feature    7702..7723
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      complement(7773..7819)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7972..7989
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(8045..8062)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(8078..8107)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_RNA        complement(8128..8203)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    8234..8252
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8259..8277
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8334..8411
                     /gene="lacI"
                     /label=lacI promoter
     CDS             8412..9035
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(8485..8504)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      9169..9200
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(9251..10129)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        10156..10440
                     /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     10363..10382
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             10467..10475
                     /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             10484..10900
                     /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             10906..11691
                     /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             11688..12368
                     /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="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      12369..12613
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
ORIGIN
        1 ttatacctag ggcgatgcag gtggctgctg aacccccagc cggaactgac cccacaaggc
       61 cctagcgttt gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc
      121 ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat
      181 ccgatccgca catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc
      241 tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga
      301 atttcgtgta gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc
      361 aacgggacgt tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt
      421 ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc
      481 attcctcggc cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct
      541 cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct
      601 gctgccacac cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca
      661 cgtccttgtt gacgtggaaa atgaccttgt ttttcagcgc ctcgcgcggg attttcttgt
      721 tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg
      781 gccacggcgc aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt
      841 tcagcaacgc ggcctgcttg gcttcgctga cctgttttgc caggtcctcg ccggcggttt
      901 ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg
      961 ccgcctcctg ttcgagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag
     1021 ggggagccag ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg actatcgagc
     1081 cgacggactg gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg
     1141 catcctcggc ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg
     1201 tccgattcat tcaccctcct tgcgggattg ccccggaatt aattccccgg atcgatccgt
     1261 cgatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt
     1321 gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt
     1381 ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct
     1441 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     1501 gcaccgtttc tgcggactgg ctttctacgt ggctgccatt tttggggtga ggccgttcgc
     1561 ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc
     1621 gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt
     1681 taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc
     1741 cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa
     1801 atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg
     1861 cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca
     1921 ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg
     1981 aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg
     2041 cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca
     2101 agttttccgc gaggtatcca caacgccggc ggccctacat ggctctgctg tagtgagtgg
     2161 gttgcgctcc ggcagcggtc ctgatccccc gcagaaaaaa aggatctcaa gaagatcctt
     2221 tgatcttttc tacactagtg cttggattct caccttggtc tgacagttac caatgcgagg
     2281 gggggatcca ccttcgggag cgcctgaagc ccgttctgga cgccctgggg ccgttgaatc
     2341 gggatatgca ggccaaggcc gccgcgatca tcaaggccgt gggcgaaaag ctgctgacgg
     2401 aacagcggga agtccagcgc cagaaacagg cccagcgcca gcaggaacgc gggcgcgcac
     2461 atttccccga aaagtgccac ctgcggcgtt gtgacaattt accgaacaac tccgcggccg
     2521 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     2581 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     2641 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     2701 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctctcc ggagactgcg agatcataga
     2761 tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa
     2821 caaccacttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     2881 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     2941 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     3001 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     3061 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     3121 atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     3181 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     3241 acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc
     3301 gtttccacgg tgtgcgtcca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     3361 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     3421 tgaattacaa cagtttttat gcatgcgcag tccagttacg ctggagtctg aggctcgtcc
     3481 tgaatgatat ctcatttatc atcatcatct ttataatcac cgctgccgtc acctcctagc
     3541 tgactcaaat caatgcgtgt ttcataaaga ccagtgatgg attgatggat aagagtggca
     3601 tctaaaactt cttttgtaga cgtatatcgt ttacgatcaa ttgttgtatc aaaatattta
     3661 aaagcagcgg gagctccaag attcgtcaac gtaaataaat gaataatatt ttctgcttgt
     3721 tcacgtattg gtttgtctct atgtttgtta tatgcactaa gaactttatc taaattggca
     3781 tctgctaaaa taacacgctt agaaaattca ctgatttgct caataatctc atctaaataa
     3841 tgcttatgct gctccacaaa caattgtttt tgttcgttat cttctggact acccttcaac
     3901 ttttcataat gactagctaa atataaaaaa ttcacatatt tgcttggcag agccagctca
     3961 tttccttttt gtaattctcc ggcactagcc agcatccgtt tacgaccgtt ttctaactca
     4021 aaaagactat atttaggtag tttaatgatt aagtcttttt taacttcctt atatccttta
     4081 gcttctaaaa agtcaatcgg atttttttca aaggaacttc tttccataat tgtgatccct
     4141 agtaactctt taacggattt taacttcttc gatttccctt tttccacctt agcaaccact
     4201 aggactgaat aagctaccgt tggactatca aaaccaccat atttttttgg atcccagtct
     4261 tttttacgag caataagctt gtccgaattt ctttttggta aaattgactc cttggagaat
     4321 ccgcctgtct gtacttctgt tttcttgaca atattgactt ggggcatgga caatactttg
     4381 cgcactgtgg caaaatctcg ccctttatcc cagacaattt ctccagtttc cccattagtt
     4441 tcgattagag ggcgtttgcg aatctctcca tttgcaagtg taatttctgt tttgaagaag
     4501 ttcatgatat tagagtaaaa gaaatatttt gcggttgctt tgcctatttc ttgctcagac
     4561 ttagcaatca ttttacgaac atcataaact ttataatcac catagacaaa ctccgattca
     4621 agttttggat atttcttaat caaagcagtt ccaacgacgg catttagata cgcatcatgg
     4681 gcatgatggt aattgttaat ctcacgtact ttatagaatt ggaaatcttt tcggaagtca
     4741 gaaactaatt tagattttaa ggtaatcact ttaacctctc gaataagttt atcattttca
     4801 tcgtatttag tattcatgcg actatccaaa atttgtgcca catgcttagt gatttggcga
     4861 gtttcaacca attggcgttt gataaaacca gctttatcaa gttcactcaa acctccacgt
     4921 tcagctttcg ttaaattatc aaacttacgt tgagtgatta acttggcgtt tagaagttgt
     4981 ctccaatagt ttttcatctt tttgactact tcttcacttg gaacgttatc cgatttacca
     5041 cgatttttat cagaacgcgt taagacctta ttgtctattg aatcgtcttt aaggaaactt
     5101 tgtggaacaa tgtgatcgac atcataatca cttaaacgat taatatctaa ttcttggtcc
     5161 acatacatgt ctcttccatt ttggagataa tagagataga gcttttcatt ttgcaattga
     5221 gtattttcaa caggatgctc tttaagaatc tgacttccta attctttgat accttcttcg
     5281 attcgtttca tacgctctcg cgaatttttc tggccctttt gagttgtctg attttcacgt
     5341 gccatttcaa taacgatatt ttctggctta tgccgcccca ttactttgac caattcatca
     5401 acaactttta cagtctgtaa aatacctttt ttaatagcag ggctaccagc taaatttgca
     5461 atatgttcat gtaaactatc gccttgtcca gacacttgtg ctttttgaat gtcttcttta
     5521 aatgtcaaac tatcatcatg gatcagctgc ataaaattgc gattggcaaa accatctgat
     5581 ttcaaaaaat ctaatattgt tttgccagat tgcttatccc taataccatt aatcaatttt
     5641 cgagacaaac gtccccaacc agtataacgg cgacgtttaa gctgtttcat caccttatca
     5701 tcaaagaggt gagcatatgt tttaagtctt tcctcaatca tctccctatc ttcaaataag
     5761 gtcaatgtta aaacaatatc ctctaagata tcttcatttt cttcattatc caaaaaatct
     5821 ttatctttaa taatttttag caaatcatgg taggtaccta atgaagcatt aaatctatct
     5881 tcaactcctg aaatttcaac actatcaaaa cattctattt ttttgaaata atcttctttt
     5941 aattgcttaa cggttacttt tcgatttgtt ttgaagagta aatcaacaat ggctttcttc
     6001 tgttcacctg aaagaaatgc tggttttcgc attccttcag taacatattt gacctttgtc
     6061 aattcgttat aaaccgtaaa atactcataa agcaaactat gttttggtag tactttttca
     6121 tttggaagat ttttatcaaa gtttgtcatg cgttcaataa atgattgagc tgaagcacct
     6181 ttatcgacaa cttcttcaaa attccatggg gtaattgttt cttcagactt ccgagtcatc
     6241 catgcaaaac gactattgcc acgcgccaat ggaccaacat aataaggaat tcgaaaagtc
     6301 aagatttttt caatcttctc acgattgtct tttaaaaatg gataaaagtc ttcttgtctt
     6361 ctcaaaatag catgcagctc acccaagtga atttgatggg gaatagagcc gttgtcaaag
     6421 gtccgttgct tgcgcagcaa atcttcacga tttagtttca ccaataattc ctcagtacca
     6481 tccatttttt ctaaaattgg tttgataaat ttataaaatt cttcttggct agctccccca
     6541 tcaatataac ctgcatatcc gttttttgat tgatcaaaaa agatttcttt atacttttct
     6601 ggaagttgtt gtcgaactaa agcttttaaa agagtcaagt cttgatgatg ttcatcgtag
     6661 cgtttaatca ttgaagctga taggggagcc ttagttattt cagtatttac tcttaggata
     6721 tctgaaagta aaatagcatc tgataaattc ttagctgcca aaaacaaatc agcatattga
     6781 tctccaattt gcgccaataa attatctaaa tcatcatcgt aagtatcttt tgaaagctgt
     6841 aatttagcat cttctgccaa atcaaaattt gatttaaaat taggggtcaa acccaatgac
     6901 aaagcaatga gattcccaaa taagccattt ttcttctcac cggggagctg agcaatgaga
     6961 ttttctaatc gtcttgattt actcaatcgt gcagaaagaa tcgctttagc atctactcca
     7021 cttgcgttaa tagggttttc ttcaaataat tgattgtagg tttgtaccaa ctggataaat
     7081 agtttgtcca catcactatt atcaggattt aaatctccct caatcaaaaa atgaccacga
     7141 aacttaatca tatgcgctaa ggccaaatag attaagcgca aatccgcttt atcagtagaa
     7201 tctaccaatt tttttcgcag atgatagata gttggatatt tctcatgata agcaacttca
     7261 tctactatat ttccaaaaat aggatgacgt tcatgcttct tgtcttcttc caccaaaaaa
     7321 gactcttcaa gtcgatgaaa gaaactatca tctactttcg ccatctcatt tgaaaaaatc
     7381 tcctgtagat aacaaatacg attcttccga cgtgtatacc ttctacgagc tgtccgtttg
     7441 agacgagtcg cttccgctgt ctctccactg tcaaataaaa gagcccctat aagatttttt
     7501 ttgatactgt ggcggtctgt atttcccaga accttgaact ttttagacgg aaccttatat
     7561 tcatcagtga tcaccgccca tccgacgcta tttgtgccga tatctaagcc tattgagtat
     7621 ttcttatcca ttgattgtac ctcctggtca gtgcgtcctg ctgatagcta gcatagtacc
     7681 taggactgag ctagccgtaa actctagaag cggccgcgaa ttcaactgca gtctagactc
     7741 gagtaaggat ccagttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     7801 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg agaccccaca
     7861 ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg tgggaccacc
     7921 gcgctactgc cgccaggcaa attctgtttt atcagaccgc ttctgcgttc tgatttaatc
     7981 tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc caagctggag accgtttaaa
     8041 ctcaatgatg atgatgatga tggtcgacgg cgctattcag atcctcttct gagatgagtt
     8101 tttgttcggg cccaagcttc aaaaaaagca ccgactcggt gccacttttt caagttgata
     8161 acggactagc cttattttaa cttgctattt ctagctctaa aacggaagct ccctcgtgcg
     8221 ctcgtgctca gtatctctat cactgatagg gatgtcaatc tctatcactg atagggactc
     8281 gagctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaactc gaggacacca
     8341 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     8401 gggtggtgaa tatgtctaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc
     8461 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag
     8521 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca
     8581 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag
     8641 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag
     8701 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat
     8761 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg
     8821 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag
     8881 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat
     8941 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat
     9001 tagaaaaaca acttaaatgt gaaagtgggt cttaatacca ccgtcaaaaa aaacggcgct
     9061 ttttagcgcc gtttttattt ttcaacctta agcttgaatt cgttgacgaa ttctctagat
     9121 atcgctcaat actgaccatt taaatcatac ctgacctcca tagcagaaag tcaaaaacct
     9181 ccgaccggag gcttttgact tgagggggat ccggcacatt tccccgaaaa gtgccacctg
     9241 catcgattta ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
     9301 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaag tagagttgat
     9361 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
     9421 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
     9481 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
     9541 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
     9601 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
     9661 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
     9721 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
     9781 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
     9841 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
     9901 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
     9961 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
    10021 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
    10081 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
    10141 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
    10201 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
    10261 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
    10321 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
    10381 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
    10441 acccgttttt ttgggaattc gagctctaag gaggttataa aaaatggata ttaatactga
    10501 aactgagatc aagcaaaagc attcactaac cccctttcct gttttcctaa tcagcccggc
    10561 atttcgcggg cgatattttc acagctattt caggagttca gccatgaacg cttattacat
    10621 tcaggatcgt cttgaggctc agagctgggc gcgtcactac cagcagctcg cccgtgaaga
    10681 gaaagaggca gaactggcag acgacatgga aaaaggcctg ccccagcacc tgtttgaatc
    10741 gctatgcatc gatcatttgc aacgccacgg ggccagcaaa aaatccatta cccgtgcgtt
    10801 tgatgacgat gttgagtttc aggagcgcat ggcagaacac atccggtaca tggttgaaac
    10861 cattgctcac caccaggttg atattgattc agaggtataa aacgaatgag tactgcactc
    10921 gcaacgctgg ctgggaagct ggctgaacgt gtcggcatgg attctgtcga cccacaggaa
    10981 ctgatcacca ctcttcgcca gacggcattt aaaggtgatg ccagcgatgc gcagttcatc
    11041 gcattactga tcgttgccaa ccagtacggc cttaatccgt ggacgaaaga aatttacgcc
    11101 tttcctgata agcagaatgg catcgttccg gtggtgggcg ttgatggctg gtcccgcatc
    11161 atcaatgaaa accagcagtt tgatggcatg gactttgagc aggacaatga atcctgtaca
    11221 tgccggattt accgcaagga ccgtaatcat ccgatctgcg ttaccgaatg gatggatgaa
    11281 tgccgccgcg aaccattcaa aactcgcgaa ggcagagaaa tcacggggcc gtggcagtcg
    11341 catcccaaac ggatgttacg tcataaagcc atgattcagt gtgcccgtct ggccttcgga
    11401 tttgctggta tctatgacaa ggatgaagcc gagcgcattg tcgaaaatac tgcatacact
    11461 gcagaacgtc agccggaacg cgacatcact ccggttaacg atgaaaccat gcaggagatt
    11521 aacactctgc tgatcgccct ggataaaaca tgggatgacg acttattgcc gctctgttcc
    11581 cagatatttc gccgcgacat tcgtgcatcg tcagaactga cacaggccga agcagtaaaa
    11641 gctcttggat tcctgaaaca gaaagccgca gagcagaagg tggcagcatg acaccggaca
    11701 ttatcctgca gcgtaccggg atcgatgtga gagctgtcga acagggggat gatgcgtggc
    11761 acaaattacg gctcggcgtc atcaccgctt cagaagttca caacgtgata gcaaaacccc
    11821 gctccggaaa gaagtggcct gacatgaaaa tgtcctactt ccacaccctg cttgctgagg
    11881 tttgcaccgg tgtggctccg gaagttaacg ctaaagcact ggcctgggga aaacagtacg
    11941 agaacgacgc cagaaccctg tttgaattca cttccggcgt gaatgttact gaatccccga
    12001 tcatctatcg cgacgaaagt atgcgtaccg cctgctctcc cgatggttta tgcagtgacg
    12061 gcaacggcct tgaactgaaa tgcccgttta cctcccggga tttcatgaag ttccggctcg
    12121 gtggtttcga ggccataaag tcagcttaca tggcccaggt gcagtacagc atgtgggtga
    12181 cgcgaaaaaa tgcctggtac tttgccaact atgacccgcg tatgaagcgt gaaggcctgc
    12241 attatgtcgt gattgagcgg gatgaaaagt acatggcgag ttttgacgag atcgtgccgg
    12301 agttcatcga aaaaatggac gaggcactgg ctgaaattgg ttttgtattt ggggagcaat
    12361 ggcgatgacg catcctcacg ataatatccg ggtaggcgca atcactttcg tctactccgt
    12421 tacaaagcga ggctgggtat ttcccggcct ttctgttatc cgaaatccac tgaaagcaca
    12481 gcggctggct gaggagataa ataataaacg aggggctgta tgcacaaagc atcttctgtt
    12541 gagttaagaa cgagtatcga gatggcacat agccttgctc aaattggaat caggtttgtg
    12601 ccaataccag tagaaacaga cgaagaatcc atgggtatgg acagttttcc ct
//
