LOCUS       sequence-434132-       12095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pKD46 with constitutively expressed Cas9 and an aTc gRNA targeting 
            the ColE1 origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12095)
  AUTHORS   Mehrer CR, Incha MR, Politz MC, Pfleger BF
  TITLE     Anaerobic production of medium-chain fatty alcohols via a 
            beta-reduction pathway.
  JOURNAL   Metab Eng. 2018 Jul;48:63-71. doi: 10.1016/j.ymben.2018.05.011. Epub
            2018 May 25.
  PUBMED    29807110
REFERENCE   2  (bases 1 to 12095)
  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: "Metab 
            Eng."; date: "2018-07"; volume: "48:63-71. doi"; pages: " 
            10.1016/j.ymben.2018.05.011. Epub 2018 May 25"
COMMENT     Sequence Label: pMP11
FEATURES             Location/Qualifiers
     source          1..12095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2..952)
                     /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      1000..1222
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(join(1844..2635,2636..2704))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2467..2486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2705..2808)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(2856..2879)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2889..6992)
                     /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(2889..6992)
                     /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(7028..7062)
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_feature    7063..7084
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      complement(7134..7180)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7333..7350
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(7406..7423)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(7439..7468)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_RNA        complement(7489..7564)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    7595..7613
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7620..7638
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7695..7772
                     /gene="lacI"
                     /label=lacI promoter
     CDS             7773..8396
                     /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(7846..7865)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      8530..8561
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(8612..9490)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        9517..9801
                     /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     9724..9743
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             9828..9836
                     /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             9845..10261
                     /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             10267..11052
                     /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             11049..11729
                     /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      11730..11974
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
ORIGIN
        1 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
       61 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
      121 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
      181 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
      241 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
      301 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
      361 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
      421 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
      481 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
      541 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
      601 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
      661 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
      721 tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca
      781 tctgaacgta ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa
      841 tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt
      901 catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca
      961 attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac
     1021 tagtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac
     1081 ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt
     1141 atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag
     1201 atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt
     1261 cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca
     1321 ccctcgcaag ctcggttgcg gccgcaatcg ggcaaatcgc tgaatattcc ttttgtctcc
     1381 gaccatcagg cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc
     1441 tctggcagtg aatgggggta aatggcacta caggcgcctt ttatggattc atgcaaggaa
     1501 actacccata atacaagaaa agcccgtcac gggcttctca gggcgtttta tggcgggtct
     1561 gctatgtggt gctatctgac tttttgctgt tcagcagttc ctgccctctg attttccagt
     1621 ctgaccactt cggattatcc cgtgacaggt cattcagact ggctaatgca cccagtaagg
     1681 cagcggtatc atcaacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     1921 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     1981 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2041 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2101 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     2161 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     2221 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     2281 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     2341 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     2401 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     2461 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     2521 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     2581 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     2641 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     2701 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2761 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcca gtccagttac
     2821 gctggagtct gaggctcgtc ctgaatgata tctcatttat catcatcatc tttataatca
     2881 ccgctgccgt cacctcctag ctgactcaaa tcaatgcgtg tttcataaag accagtgatg
     2941 gattgatgga taagagtggc atctaaaact tcttttgtag acgtatatcg tttacgatca
     3001 attgttgtat caaaatattt aaaagcagcg ggagctccaa gattcgtcaa cgtaaataaa
     3061 tgaataatat tttctgcttg ttcacgtatt ggtttgtctc tatgtttgtt atatgcacta
     3121 agaactttat ctaaattggc atctgctaaa ataacacgct tagaaaattc actgatttgc
     3181 tcaataatct catctaaata atgcttatgc tgctccacaa acaattgttt ttgttcgtta
     3241 tcttctggac tacccttcaa cttttcataa tgactagcta aatataaaaa attcacatat
     3301 ttgcttggca gagccagctc atttcctttt tgtaattctc cggcactagc cagcatccgt
     3361 ttacgaccgt tttctaactc aaaaagacta tatttaggta gtttaatgat taagtctttt
     3421 ttaacttcct tatatccttt agcttctaaa aagtcaatcg gatttttttc aaaggaactt
     3481 ctttccataa ttgtgatccc tagtaactct ttaacggatt ttaacttctt cgatttccct
     3541 ttttccacct tagcaaccac taggactgaa taagctaccg ttggactatc aaaaccacca
     3601 tatttttttg gatcccagtc ttttttacga gcaataagct tgtccgaatt tctttttggt
     3661 aaaattgact ccttggagaa tccgcctgtc tgtacttctg ttttcttgac aatattgact
     3721 tggggcatgg acaatacttt gcgcactgtg gcaaaatctc gccctttatc ccagacaatt
     3781 tctccagttt ccccattagt ttcgattaga gggcgtttgc gaatctctcc atttgcaagt
     3841 gtaatttctg ttttgaagaa gttcatgata ttagagtaaa agaaatattt tgcggttgct
     3901 ttgcctattt cttgctcaga cttagcaatc attttacgaa catcataaac tttataatca
     3961 ccatagacaa actccgattc aagttttgga tatttcttaa tcaaagcagt tccaacgacg
     4021 gcatttagat acgcatcatg ggcatgatgg taattgttaa tctcacgtac tttatagaat
     4081 tggaaatctt ttcggaagtc agaaactaat ttagatttta aggtaatcac tttaacctct
     4141 cgaataagtt tatcattttc atcgtattta gtattcatgc gactatccaa aatttgtgcc
     4201 acatgcttag tgatttggcg agtttcaacc aattggcgtt tgataaaacc agctttatca
     4261 agttcactca aacctccacg ttcagctttc gttaaattat caaacttacg ttgagtgatt
     4321 aacttggcgt ttagaagttg tctccaatag tttttcatct ttttgactac ttcttcactt
     4381 ggaacgttat ccgatttacc acgattttta tcagaacgcg ttaagacctt attgtctatt
     4441 gaatcgtctt taaggaaact ttgtggaaca atgtgatcga catcataatc acttaaacga
     4501 ttaatatcta attcttggtc cacatacatg tctcttccat tttggagata atagagatag
     4561 agcttttcat tttgcaattg agtattttca acaggatgct ctttaagaat ctgacttcct
     4621 aattctttga taccttcttc gattcgtttc atacgctctc gcgaattttt ctggcccttt
     4681 tgagttgtct gattttcacg tgccatttca ataacgatat tttctggctt atgccgcccc
     4741 attactttga ccaattcatc aacaactttt acagtctgta aaataccttt tttaatagca
     4801 gggctaccag ctaaatttgc aatatgttca tgtaaactat cgccttgtcc agacacttgt
     4861 gctttttgaa tgtcttcttt aaatgtcaaa ctatcatcat ggatcagctg cataaaattg
     4921 cgattggcaa aaccatctga tttcaaaaaa tctaatattg ttttgccaga ttgcttatcc
     4981 ctaataccat taatcaattt tcgagacaaa cgtccccaac cagtataacg gcgacgttta
     5041 agctgtttca tcaccttatc atcaaagagg tgagcatatg ttttaagtct ttcctcaatc
     5101 atctccctat cttcaaataa ggtcaatgtt aaaacaatat cctctaagat atcttcattt
     5161 tcttcattat ccaaaaaatc tttatcttta ataattttta gcaaatcatg gtaggtacct
     5221 aatgaagcat taaatctatc ttcaactcct gaaatttcaa cactatcaaa acattctatt
     5281 tttttgaaat aatcttcttt taattgctta acggttactt ttcgatttgt tttgaagagt
     5341 aaatcaacaa tggctttctt ctgttcacct gaaagaaatg ctggttttcg cattccttca
     5401 gtaacatatt tgacctttgt caattcgtta taaaccgtaa aatactcata aagcaaacta
     5461 tgttttggta gtactttttc atttggaaga tttttatcaa agtttgtcat gcgttcaata
     5521 aatgattgag ctgaagcacc tttatcgaca acttcttcaa aattccatgg ggtaattgtt
     5581 tcttcagact tccgagtcat ccatgcaaaa cgactattgc cacgcgccaa tggaccaaca
     5641 taataaggaa ttcgaaaagt caagattttt tcaatcttct cacgattgtc ttttaaaaat
     5701 ggataaaagt cttcttgtct tctcaaaata gcatgcagct cacccaagtg aatttgatgg
     5761 ggaatagagc cgttgtcaaa ggtccgttgc ttgcgcagca aatcttcacg atttagtttc
     5821 accaataatt cctcagtacc atccattttt tctaaaattg gtttgataaa tttataaaat
     5881 tcttcttggc tagctccccc atcaatataa cctgcatatc cgttttttga ttgatcaaaa
     5941 aagatttctt tatacttttc tggaagttgt tgtcgaacta aagcttttaa aagagtcaag
     6001 tcttgatgat gttcatcgta gcgtttaatc attgaagctg ataggggagc cttagttatt
     6061 tcagtattta ctcttaggat atctgaaagt aaaatagcat ctgataaatt cttagctgcc
     6121 aaaaacaaat cagcatattg atctccaatt tgcgccaata aattatctaa atcatcatcg
     6181 taagtatctt ttgaaagctg taatttagca tcttctgcca aatcaaaatt tgatttaaaa
     6241 ttaggggtca aacccaatga caaagcaatg agattcccaa ataagccatt tttcttctca
     6301 ccggggagct gagcaatgag attttctaat cgtcttgatt tactcaatcg tgcagaaaga
     6361 atcgctttag catctactcc acttgcgtta atagggtttt cttcaaataa ttgattgtag
     6421 gtttgtacca actggataaa tagtttgtcc acatcactat tatcaggatt taaatctccc
     6481 tcaatcaaaa aatgaccacg aaacttaatc atatgcgcta aggccaaata gattaagcgc
     6541 aaatccgctt tatcagtaga atctaccaat ttttttcgca gatgatagat agttggatat
     6601 ttctcatgat aagcaacttc atctactata tttccaaaaa taggatgacg ttcatgcttc
     6661 ttgtcttctt ccaccaaaaa agactcttca agtcgatgaa agaaactatc atctactttc
     6721 gccatctcat ttgaaaaaat ctcctgtaga taacaaatac gattcttccg acgtgtatac
     6781 cttctacgag ctgtccgttt gagacgagtc gcttccgctg tctctccact gtcaaataaa
     6841 agagccccta taagattttt tttgatactg tggcggtctg tatttcccag aaccttgaac
     6901 tttttagacg gaaccttata ttcatcagtg atcaccgccc atccgacgct atttgtgccg
     6961 atatctaagc ctattgagta tttcttatcc attgattgta cctcctggtc agtgcgtcct
     7021 gctgatagct agcatagtac ctaggactga gctagccgta aactctagaa gcggccgcga
     7081 attcaactgc agtctagact cgagtaagga tccagttcac cgacaaacaa cagataaaac
     7141 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca gttccctact
     7201 ctcgcatggg gagaccccac actaccatcg gcgctacggc gtttcacttc tgagttcggc
     7261 atggggtcag gtgggaccac cgcgctactg ccgccaggca aattctgttt tatcagaccg
     7321 cttctgcgtt ctgatttaat ctgtatcagg ctgaaaatct tctctcatcc gccaaaacag
     7381 ccaagctgga gaccgtttaa actcaatgat gatgatgatg atggtcgacg gcgctattca
     7441 gatcctcttc tgagatgagt ttttgttcgg gcccaagctt caaaaaaagc accgactcgg
     7501 tgccactttt tcaagttgat aacggactag ccttatttta acttgctatt tctagctcta
     7561 aaacggaagc tccctcgtgc gctcgtgctc agtatctcta tcactgatag ggatgtcaat
     7621 ctctatcact gatagggact cgagctatgc tactccgtca agccgtcaat tgtctgattc
     7681 gttaccaact cgaggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     7741 gcccggaaga gagtcaattc agggtggtga atatgtctag attagataaa agtaaagtga
     7801 ttaacagcgc attagagctg cttaatgagg tcggaatcga aggtttaaca acccgtaaac
     7861 tcgcccagaa gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg
     7921 ctttgctcga cgccttagcc attgagatgt tagataggca ccatactcac ttttgccctt
     7981 tagaagggga aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt
     8041 tactaagtca tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt
     8101 atgaaactct cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg
     8161 cattatatgc actcagcgct gtggggcatt ttactttagg ttgcgtattg gaagatcaag
     8221 agcatcaagt cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat
     8281 tacgacaagc tatcgaatta tttgatcacc aaggtgcaga gccagccttc ttattcggcc
     8341 ttgaattgat catatgcgga ttagaaaaac aacttaaatg tgaaagtggg tcttaatacc
     8401 accgtcaaaa aaaacggcgc tttttagcgc cgtttttatt tttcaacctt aagcttgaat
     8461 tcgttgacga attctctaga tatcgctcaa tactgaccat ttaaatcata cctgacctcc
     8521 atagcagaaa gtcaaaaacc tccgaccgga ggcttttgac ttgaggggga tccggcacat
     8581 ttccccgaaa agtgccacct gcatcgattt attatgacaa cttgacggct acatcattca
     8641 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     8701 cccgcgagaa gtagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     8761 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     8821 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     8881 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     8941 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     9001 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     9061 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     9121 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     9181 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     9241 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     9301 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     9361 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     9421 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     9481 cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca
     9541 tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc
     9601 cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca
     9661 aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt
     9721 gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc
     9781 aactctctac tgtttctcca tacccgtttt tttgggaatt cgagctctaa ggaggttata
     9841 aaaaatggat attaatactg aaactgagat caagcaaaag cattcactaa ccccctttcc
     9901 tgttttccta atcagcccgg catttcgcgg gcgatatttt cacagctatt tcaggagttc
     9961 agccatgaac gcttattaca ttcaggatcg tcttgaggct cagagctggg cgcgtcacta
    10021 ccagcagctc gcccgtgaag agaaagaggc agaactggca gacgacatgg aaaaaggcct
    10081 gccccagcac ctgtttgaat cgctatgcat cgatcatttg caacgccacg gggccagcaa
    10141 aaaatccatt acccgtgcgt ttgatgacga tgttgagttt caggagcgca tggcagaaca
    10201 catccggtac atggttgaaa ccattgctca ccaccaggtt gatattgatt cagaggtata
    10261 aaacgaatga gtactgcact cgcaacgctg gctgggaagc tggctgaacg tgtcggcatg
    10321 gattctgtcg acccacagga actgatcacc actcttcgcc agacggcatt taaaggtgat
    10381 gccagcgatg cgcagttcat cgcattactg atcgttgcca accagtacgg ccttaatccg
    10441 tggacgaaag aaatttacgc ctttcctgat aagcagaatg gcatcgttcc ggtggtgggc
    10501 gttgatggct ggtcccgcat catcaatgaa aaccagcagt ttgatggcat ggactttgag
    10561 caggacaatg aatcctgtac atgccggatt taccgcaagg accgtaatca tccgatctgc
    10621 gttaccgaat ggatggatga atgccgccgc gaaccattca aaactcgcga aggcagagaa
    10681 atcacggggc cgtggcagtc gcatcccaaa cggatgttac gtcataaagc catgattcag
    10741 tgtgcccgtc tggccttcgg atttgctggt atctatgaca aggatgaagc cgagcgcatt
    10801 gtcgaaaata ctgcatacac tgcagaacgt cagccggaac gcgacatcac tccggttaac
    10861 gatgaaacca tgcaggagat taacactctg ctgatcgccc tggataaaac atgggatgac
    10921 gacttattgc cgctctgttc ccagatattt cgccgcgaca ttcgtgcatc gtcagaactg
    10981 acacaggccg aagcagtaaa agctcttgga ttcctgaaac agaaagccgc agagcagaag
    11041 gtggcagcat gacaccggac attatcctgc agcgtaccgg gatcgatgtg agagctgtcg
    11101 aacaggggga tgatgcgtgg cacaaattac ggctcggcgt catcaccgct tcagaagttc
    11161 acaacgtgat agcaaaaccc cgctccggaa agaagtggcc tgacatgaaa atgtcctact
    11221 tccacaccct gcttgctgag gtttgcaccg gtgtggctcc ggaagttaac gctaaagcac
    11281 tggcctgggg aaaacagtac gagaacgacg ccagaaccct gtttgaattc acttccggcg
    11341 tgaatgttac tgaatccccg atcatctatc gcgacgaaag tatgcgtacc gcctgctctc
    11401 ccgatggttt atgcagtgac ggcaacggcc ttgaactgaa atgcccgttt acctcccggg
    11461 atttcatgaa gttccggctc ggtggtttcg aggccataaa gtcagcttac atggcccagg
    11521 tgcagtacag catgtgggtg acgcgaaaaa atgcctggta ctttgccaac tatgacccgc
    11581 gtatgaagcg tgaaggcctg cattatgtcg tgattgagcg ggatgaaaag tacatggcga
    11641 gttttgacga gatcgtgccg gagttcatcg aaaaaatgga cgaggcactg gctgaaattg
    11701 gttttgtatt tggggagcaa tggcgatgac gcatcctcac gataatatcc gggtaggcgc
    11761 aatcactttc gtctactccg ttacaaagcg aggctgggta tttcccggcc tttctgttat
    11821 ccgaaatcca ctgaaagcac agcggctggc tgaggagata aataataaac gaggggctgt
    11881 atgcacaaag catcttctgt tgagttaaga acgagtatcg agatggcaca tagccttgct
    11941 caaattggaa tcaggtttgt gccaatacca gtagaaacag acgaagaatc catgggtatg
    12001 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
    12061 gatgcttcac tgatagatac aagagccata agaac
//
