pCfB6364

LOCUS      pCfB6364     16432 bp    DNA     circular SYN 14-APR-2025
DEFINITION  EasyCloneYALI system-based yeast integrative vector expressing Cas9 
            carrying D-serine marker, integration into Yarrowia lipolytica ku70 
            locus, amp resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16432)
  AUTHORS   Holkenbrink C, Dam MI, Kildegaard KR, Beder J, Dahlin J, Domenech 
            Belda D, Borodina I
  TITLE     EasyCloneYALI: CRISPR/Cas9-Based Synthetic Toolbox for Engineering 
            of the Yeast Yarrowia lipolytica.
  JOURNAL   Biotechnol J. 2018 Jan 29. doi: 10.1002/biot.201700543.
  PUBMED    29377615
REFERENCE   2  (bases 1 to 16432)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            J. 2018 Jan 29. doi: 10.1002/biot.201700543."
COMMENT     Sequence Label: pCfB6364
FEATURES             Location/Qualifiers
     source          1..16432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(32..65)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(272..291)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(297..316)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        358..449
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(450..518,519..1310)
                     /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     complement(668..687)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1432..2020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1921..1940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2174..2191
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2289..2306
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2290..2306
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2776..2809
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             5601..9704
                     /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             5601..9704
                     /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             9717..9737
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    11123..11156
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    14355..14388
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cattcgcggc cgcatttaaa tccccacttc agaagttcct atactttcta gagaatagga
       61 acttcactat agagtcgaat aagggcgaca ccccataatt agcccgggcg aaaggcccag
      121 tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg
      181 agtccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg
      241 tgggaccacc gcgctactgc cgccaggcaa acaaggggtg ttatgagcca tattcaggta
      301 taaatgggct cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt
      361 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      421 tgcttcaata atattgaaaa aggaagaata tgagtattca acatttccgt gtcgccctta
      481 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      541 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
      601 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
      661 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
      721 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
      781 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
      841 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
      901 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
      961 taccaaacga cgagcgtgac accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac
     1021 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1081 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1141 ataaatccgg agccggtgag cgtggttctc gcggtatcat cgcagcgctg gggccagatg
     1201 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1261 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa gcagagcatt
     1321 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     1381 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1441 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1501 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1561 gcagatacca aatactgttc ttctagtgta gccgtagtta gcccaccact tcaagaactc
     1621 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1681 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1741 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1801 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1861 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1921 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1981 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2041 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2101 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2161 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa ggcgagagta gggaactgcc
     2221 aggcatcaaa ctaagcagaa ggcccctgac ggatggcctt tttgcgtttc tacaaactct
     2281 ttctgtgttg taaaacgacg gccagtctta agctcgggcc ccctgggcgg ttctgataac
     2341 gagtaatcgt taatccgcaa ataacgtaaa aacccgcttc ggcgggtttt tttatggggg
     2401 gagtttaggg aaagagcatt tgtcagaata tttaagggcg cctgtcactt tgcttgatat
     2461 atgagaatta tttaacctta taaatgagaa aaaagcaacg cactttaaat aagatacgtt
     2521 gctttttcga ttgatgaaca cctataatta aactattcat ctattattta tgattttttg
     2581 tatatacaat atttctagtt tgttaaagag aattaagaaa ataaatctcg aaaataataa
     2641 agggaaaatc agtttttgat atcaaaatta tacatgtcaa cgataataca aaatataata
     2701 caaactataa gatgttatca gtatttatta tgcatttaga ataaattttg tgtcgccctt
     2761 attcgactca ctatagaagt tcctattctc tagaaagtat aggaacttca cttcatttta
     2821 tttaaatttg cggccgcgaa tgcgtgcgat atggtgtgcc aggaggtgga cgcagagaaa
     2881 tacaactggt ggaacgagtt tctcagcaaa catgggtacg acagtgtctt tagcacatac
     2941 gagggaaaga accatggact catgtgtgcc tggaagaagg acatgtttga atttgaaaac
     3001 aaacaggcaa ttgagtacga ccatgtgatc atggagaacc tgcctgtgca gttcaagacc
     3061 cgaaacacag gtctgattgt gcagttgaaa catgctgtca cagccaagac gattatcgtg
     3121 gcgaccactc atctgttctg gcatcccatg ggctcatttg aacgtaccaa acaaactgcc
     3181 atgttgttac gtgcatctca gaagtacgtg gaggatttgg atgccgagaa gggcacgaaa
     3241 tccattcttg tggtagctgg agatttcaac tccactccct gggacggtcc ctacgtcgga
     3301 gctaccatgc acaagttcga cgagattacg cggcccattt taagtgcttc tctgcagtac
     3361 aagttcactc gaaagagcaa ggagcaaaag gctgctgaag ctgcaaaggc agacgcgatt
     3421 gcaaagggag aacctcttcc tgaggagcct gaacccgaag aagacgaggg ggatggcgag
     3481 ttagaagctc taggggacgt tgtcatttca gaggaggatg cagaggccca cattcagacc
     3541 cttgagaatg aattcagaaa gctgcctgta ctcaactctc tgtatggcag ctacacagat
     3601 gttgttcctg aagccaccaa ggagcccgtt ggcaagcaca agaaggagcc tcgcttcagt
     3661 aactgggccc acgcctggag gggtcttttg gactacattt tcgtttcgtc caacaacagc
     3721 gatgaggtcc aggtcaagag tctgttgaag atgccccttg aggaggatat ggggcccgag
     3781 cccagtggtc agcctcgcat tgacatgtat ccttcagatc atcttgctat catggctgaa
     3841 gttgagctga agtgaacgac caagactaaa gggtggcaca taaatggaca aggtatatag
     3901 aaacgtgata aaggaggccg caaaggttct attggcttag tgccgagttt ggtggtccat
     3961 tgttcacttg atggtatgta ctgtactgta gctacaatag agcagttgac gatatagccc
     4021 tttgtagtgc tctatagact attgtgtgtc gtaacattgg agtgccaacc tacctctagc
     4081 aacacactca ttcacaagag agaaacgaca cagaagctgt tgttaatcag gcagcaggta
     4141 ttccattgcc ctttgctctc gttaagtcat ctgatttcac tcgctacttc ttactgagca
     4201 aatacaaccc tcttcatcgg tacattatac aaagtcatcc accactagcc actactttcg
     4261 ccaccaagat gggcaagaag aaccgcttca aaaaggtgag tcgagaactt cattttggcc
     4321 gcaagttgga gcaatcacct actctctctc tcgacggaga ggctcaattg gagcggtttg
     4381 ctcgaaacca gtggtaccgc tactttgtgc agattgaggc caagtaccga atggtatcca
     4441 acgacgatga gatcacggct gcccaccaaa ccatgatgtt gcttggacaa cctgttggca
     4501 tttccctgga ctccgagctc ggtcagaaga ttgaggctga gctgcaggga caacgacagg
     4561 aacaggaggc ggacgctcat ccagtggaaa ctgtcaaccc gcccagaaga ccatggagac
     4621 ggtcaagaaa ggcgtagcta cggtgtaact tgggccaaac cacgtgacag tcacatgatt
     4681 caccgctcat acgacatcgc acagacacca gtatatagat aatatatttg tactctatct
     4741 taatctgaaa gaggactcat cctcatatcg tttggaagca gcagtcatac atgttctgag
     4801 atatgatggt atcattaata ttcgttattc gtttagatgt gcctccctag acgtgcaacg
     4861 cttacgcaac taacatgaat gaatacgata tacatcaaag actatgatac gcagtattgc
     4921 acactgtacg agtaagggca ctagccactg cactcaagtg aaaccgttgc ccgggtacga
     4981 gtatgagtat gtacagtatg tttagtattg tacttggaca atgcttgtat cgtacattct
     5041 caagtgtcaa acataaatat ccgttgctat atcctcgcac caccacgtag ctcgctatat
     5101 ccctgtgttg aatccatcca tcttggattg ccaattgtgc acacagaacc gggcactcac
     5161 ttccccatcc acacttcaac ggaatgcgtg cgatagagac cgggttggcg gcgcatttgt
     5221 gtcccaaaaa acagccccaa ttgccccaat tgaccccaaa ttgacccagt agcgggccca
     5281 accccggcga gagccccctt ctccccacat atcaaacctc ccccggttcc cacacttgcc
     5341 gttaagggcg tagggtactg cagtctggaa tctacgcttg ttcagacttt gtactagttt
     5401 ctttgtctgg ccatccgggt aacccatgcc ggacgcaaaa tagactactg aaaatttttt
     5461 tgctttgtgg ttgggacttt agccaagggt ataaaagacc accgtccccg aattaccttt
     5521 cctcttcttt tctctctctc cttgtcaact cacacccgaa atcgttaagc atttccttct
     5581 gagtataaga atcattcaaa atggacaaga agtactctat cggcctggac atcggcacca
     5641 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gccctctaag aagttcaagg
     5701 tcctgggcaa caccgaccga cactctatca agaagaacct gatcggcgcc ctgctgttcg
     5761 actctggcga gactgccgag gccacccgac tgaagcgaac cgcccgacga cgatacaccc
     5821 gacgaaagaa ccgaatctgc tacctgcaag agatcttctc taacgagatg gccaaggtgg
     5881 acgactcgtt cttccaccga ctcgaggaat ctttcctggt cgaggaagat aagaagcacg
     5941 agcgacaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     6001 ccatctacca cctccgaaag aagctggttg actctaccga caaggccgac ctgcgactga
     6061 tctacctggc cctggcccac atgatcaagt tccgaggcca cttcctgatc gagggcgacc
     6121 tgaaccccga caactctgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     6181 agctgttcga ggaaaacccc atcaacgcct ctggcgtgga cgccaaggcc atcctgtctg
     6241 cccgactgtc taagtctcga cgactcgaga acctgattgc ccagctgccc ggcgagaaga
     6301 agaacggcct gttcggcaac ctgatcgccc tgtctctggg cctgaccccc aacttcaagt
     6361 ctaacttcga cctggccgag gacgccaagc tgcagctgtc taaggacacc tacgacgacg
     6421 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttc ctggccgcca
     6481 agaacctgtc tgacgccatc ctgctgtctg acatcctgcg agtgaacacc gagatcacca
     6541 aggcccccct gtccgcctct atgatcaagc gatacgacga gcaccaccag gacctgaccc
     6601 tgctgaaggc cctggtgcga cagcagctgc ccgagaagta caaagagatt ttcttcgacc
     6661 agtctaagaa cggctacgcc ggctacatcg acggcggagc ctctcaagag gaattctaca
     6721 agttcatcaa gcccatcctc gagaagatgg acggcaccga ggaactgctg gtgaagctga
     6781 accgagagga cctgctgcga aagcagcgaa ccttcgacaa cggctctatc ccccaccaga
     6841 tccacctggg cgagctgcac gccattctgc gacgacaaga ggacttctac cccttcctga
     6901 aagacaaccg agagaagatc gagaagattc tgaccttccg aatcccctac tacgtgggcc
     6961 ccctggcccg aggcaactct cgattcgcct ggatgacccg aaagtctgag gaaactatca
     7021 ccccctggaa cttcgaggaa gtggtggaca agggcgcctc tgcccagtct ttcatcgagc
     7081 gaatgaccaa cttcgacaag aacctgccca acgagaaggt gctgcccaag cactctctgc
     7141 tgtacgagta cttcaccgtg tacaacgagc tgaccaaggt gaagtacgtg accgagggaa
     7201 tgcgaaagcc cgccttcctg tctggcgagc agaagaaggc catcgttgac ctgctgttca
     7261 agaccaaccg aaaggtgacc gtgaagcagc tgaaagagga ctacttcaaa aagatcgagt
     7321 gcttcgactc cgtcgagatc tctggcgtcg aggaccgatt caacgcctct ctgggcacct
     7381 accacgacct gctgaagatt atcaaggaca aggacttcct ggacaacgag gaaaacgagg
     7441 acatcctcga ggacatcgtg ctgaccctga ccctgttcga ggaccgagag atgattgagg
     7501 aacgactcaa gacctacgcc cacctgttcg acgacaaggt gatgaagcag ctcaagcgac
     7561 gacgatacac tggctggggc cgactgtctc gaaagctgat caacggcatc cgagacaagc
     7621 agtctggcaa gaccatcctg gactttctga agtctgacgg cttcgccaac cgaaacttca
     7681 tgcagctgat ccacgacgac tccctgacct tcaaagagga catccagaag gcccaggttt
     7741 ctggccaggg cgactctctg cacgagcaca ttgccaacct ggccggctct cccgccatca
     7801 agaagggcat cctgcagacc gtgaaggtgg tggacgagct ggtgaaggtg atgggccgac
     7861 acaagcccga gaacatcgtg atcgagatgg cccgagagaa ccagaccacc cagaagggcc
     7921 agaagaactc tcgagagcga atgaagcgaa tcgaggaagg catcaaagaa ctgggctctc
     7981 agatcctgaa agaacacccc gtcgagaaca cccagctgca gaacgagaag ctgtacctgt
     8041 actacctgca gaacggccga gacatgtacg tggaccaaga actggacatc aaccgactgt
     8101 ctgactacga cgtggaccat atcgtgcccc agtcttttct gaaggacgac tctatcgaca
     8161 acaaggtgct gacccgatct gacaagaacc gaggcaagtc tgacaacgtg ccctctgaag
     8221 aagtggtcaa gaagatgaag aactactggc gacagctgct gaacgccaag ctgatcaccc
     8281 agcgaaagtt cgacaacctg accaaggccg agcgaggcgg cctgtctgag ctggacaagg
     8341 ccggcttcat caagcgacag ctggtcgaga ctcgacagat caccaagcac gtggcccaga
     8401 tcctggactc tcgaatgaac accaagtacg acgagaacga caagctgatt cgagaggtga
     8461 aggtcatcac cctgaagtct aagctggtgt ctgacttccg aaaggacttc cagttctaca
     8521 aggtgcgaga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtggtgg
     8581 gcaccgccct gatcaagaag taccctaagc tcgagtctga gttcgtgtac ggcgactaca
     8641 aggtgtacga cgtgcgaaag atgatcgcca agtctgagca agagatcggc aaggctaccg
     8701 ccaagtactt cttctactct aacatcatga actttttcaa gaccgagatt accctggcca
     8761 acggcgagat ccgaaagcga cccctgatcg agactaacgg cgagactggc gagatcgtgt
     8821 gggacaaggg ccgagacttc gccaccgtgc gaaaggtgct gtctatgccc caggtgaaca
     8881 tcgtgaagaa gaccgaggtc cagaccggcg gcttctctaa agagtctatc ctgcccaagc
     8941 gaaactctga caagctgatc gcccgaaaga aggactggga ccccaagaag tacggcggct
     9001 tcgactctcc caccgtggcc tactctgtgc tggtggtggc caaggtcgag aagggcaagt
     9061 ctaagaagct gaagtctgtc aaagagctgc tcggcatcac tatcatggaa cgatcttcgt
     9121 tcgagaagaa ccctatcgac ttcctcgagg ccaagggcta caaagaggtg aagaaggacc
     9181 tgatcatcaa gctgcccaag tactccctgt tcgagctgga gaacggccga aagcgaatgc
     9241 tggcctctgc cggcgagctg cagaagggca acgagctggc cctgccctct aagtacgtga
     9301 acttcctgta cctggcctct cactacgaga agctgaaggg ctctcccgag gacaacgagc
     9361 agaagcagct gttcgtcgag cagcacaagc actacctgga cgagatcatc gagcagatct
     9421 ctgaattctc taagcgagtg atcctggccg acgccaacct ggacaaggtg ctgtccgcct
     9481 acaacaagca ccgagacaag cccatccgag agcaggccga gaacatcatc cacctgttca
     9541 ccctgaccaa cctgggagcc cccgctgcct tcaagtactt cgacaccacc atcgaccgaa
     9601 agcgatacac ctctaccaaa gaggtgctgg acgccaccct gatccaccag tctatcaccg
     9661 gcctgtacga gactcgaatc gacctgtctc agctgggcgg cgactctcga gccgacccta
     9721 agaagaagcg aaaggtctaa ccccacgttg ccggtcttgc ctcctactac ctgtccatca
     9781 atgacgaggt tctcacccct gcccaggtcg aggctcttat tactgagtcc aacaccggtg
     9841 ttcttcccac caccaacctc aagggctctc ccaacgctgt tgcctacaac ggtgttggca
     9901 tttaggcaat taacagatag tttgccggtg ataattctct taacctccca cactcctttg
     9961 acataacgat ttatgtaacg aaactgaaat ttgaccagat attgttgtaa atagaaaatc
    10021 tggcttgtag gtggcaaaat cccgtctttg ttcatcaatt ccctctgtga ctactcgtca
    10081 tccctttatg ttcgactgtc gtatttttat tttccataca tacgcaagtg agatgcccgt
    10141 gtccgaattc atcgcgtgca ttccttctgt tcggaatcaa cctcaaggtt aacggccacg
    10201 atcccctcgt tgttactctt ggtcagccca ttgtcggtaa cgctggcttt gctaactggg
    10261 tcgataaact cttctttggc caggagaacc ccgatgtctc caaggtgtcc aaagaccgaa
    10321 agctctaccg aatcacccac cgaggagata tcgtccctca agtgcccttc tgggacggtt
    10381 accagcactg ctctggtgag gtctttattg actggcccct gatccaccct cctctctcca
    10441 acgttgtcat gtgccagggc cagagcaata aacagtgctc tgccggtaac actctgctcc
    10501 agcaggtcaa tgtgattgga aaccatctgc agtacttcgt caccgagggt gtctgtggta
    10561 tctaagctat ttatcactct ttacaacttc tacctcaact atctacttta ataaatgaat
    10621 atcgtttatt ctctatgatt actgtatatg cgttcctcta agacaaatcg aaaccagcat
    10681 gcgatcgaat ggcatacaaa agtttcttcc gaagttgatc aatgtcctga tagtcaggca
    10741 gcttgagaag attgacacag gtggaggccg tagggaaccg atcaacctgt ctaccagcgt
    10801 tacgaatggc aaatgacggg ttcaaagcct tgaatccttg caatggtgcc ttggatactg
    10861 atgtcacaaa cttaagaagc agccgcttgt cctcttcctc gaaactctca aacacagtcc
    10921 agaagtcctt tatagtttga tctgtatcca gatagcctcc gtaattggtg tgtgtcttca
    10981 aatcccagac gtccacattg gcatgtcctc cactgataag catttgaagt tcatctgcgt
    11041 tgaacattga gacccacgaa gggtcaatga gctggtatag accgcccaag aatgcatctg
    11101 agaacagctg aagcttcgta cgataacttc gtataatgta tgctatacga agttatgacg
    11161 cagtaggatg tcctgcacgg gtctttttgt ggggtgtgga gaaaggggtg cttggagatg
    11221 gaagccggta gaaccgggct gcttgggggg atttggggcc gctgggctcc aaagaggggt
    11281 aggcatttcg ttggggttac gtaattgcgg catttgggtc ctgcgcgcat gtcccattgg
    11341 tcagaattag tccggatagg agacttatca gccaatcaca gcgccggatc cacctgtagg
    11401 ttgggttggg tgggagcacc cctccacaga gtagagtcaa acagcagcag caacatgata
    11461 gttgggggtg tgcgtgttaa aggaaaaaaa aagaagcttg ggttatattc ccgctctatt
    11521 tagaggttgc gggatagacg ccgacggagg gcaatggcgc catggaacct tgcggatatc
    11581 gatacgccgc ggcggactgc gtccgaacca gctccagcag cgttttttcc gggccattga
    11641 gccgactgcg accccgccaa cgtgtcttgg cccacgcact catgtcatgt tggtgttggg
    11701 aggccacttt ttaagtagca caaggcacct agctcgcagc aaggtgtccg aaccaaagaa
    11761 gcggctgcag tggtgcaaac ggggcggaaa cggcgggaaa aagccacggg ggcacgaatt
    11821 gaggcacgcc ctcgaatttg agacgagtca cggccccatt cgcccgcgca atggctcgcc
    11881 aacgcccggt cttttgcacc acatcaggtt accccaagcc aaacctttgt gttaaaaagc
    11941 ttaacatatt ataccgaacg taggtttggg cgggcttgct ccgtctgtcc aaggcaacat
    12001 ttatataagg gtctgcatcg ccggctcaat tgaatctttt ttcttcttct cttctctata
    12061 ttcattcttg aattaaacac acatcaacat ggaaaacgct aaaatgaact cgctcatcgc
    12121 ccagtatccg ttggtaaagg atctggttgc tcttaaagaa accacctggt ttaatcctgg
    12181 cacgacctca ttggctgaag gtttacctta tgttggcctg accgaacagg atgttcagga
    12241 cgcccatgcg cgcttatccc gttttgcacc ctatctggca aaagcatttc ctgaaactgc
    12301 tgccactggg gggattattg aatcagaact ggttgccatt ccagctatgc aaaaacggct
    12361 ggaaaaagaa tatcagcaac cgatcagcgg gcaactgtta ctgaaaaaag atagccattt
    12421 gcccatttcc ggctccataa aagcacgcgg cgggatttat gaagtcctgg cacacgcaga
    12481 aaaactggct ctggaagcgg ggttgctgac gcttgatgat gactacagca aactgctttc
    12541 tccggagttt aaacagttct ttagccaata cagcattgct gtgggctcaa ccggaaatct
    12601 ggggttatca atcggcatta tgagcgcccg cattggcttt aaggtgacag ttcatatgtc
    12661 tgctgatgcc cgggcatgga aaaaagcgaa actgcgcagc catggcgtta cggtcgtgga
    12721 atatgagcaa gattatggtg ttgccgtcga ggaaggacgt aaagcagcgc agtctgaccc
    12781 gaactgtttc tttattgatg acgaaaattc ccgcacgttg ttccttgggt attccgtcgc
    12841 tggccagcgt cttaaagcgc aatttgccca gcaaggccgt atcgtcgatg ctgataaccc
    12901 tctgtttgtc tatctgccgt gtggtgttgg cggtggtcct ggtggcgtcg ctttcgggct
    12961 taaactggcg tttggcgatc atgttcactg cttttttgcc gaaccaacgc actccccttg
    13021 tatgttgtta ggcgtccata caggattaca cgatcagatt tctgttcagg atattggtat
    13081 cgacaacctt accgcagcgg atggccttgc agttggtcgc gcatcaggct ttgtcgggcg
    13141 ggcaatggag cgtctgctgg atggcttcta tacccttagc gatcaaacca tgtatgacat
    13201 gcttggctgg ctggcgcagg aagaaggtat tcgtcttgaa ccttcggcac tggcgggtat
    13261 ggccggacct cagcgcgtgt gtgcatcagt aagttaccaa cagatgcacg gtttcagcgc
    13321 agaacaactg cgtaatacca ctcatctggt gtgggcgacg ggaggtggaa tggtgccgga
    13381 agaagagatg aatcaatatc tggcaaaagg ccgttaactt ctgttcggaa tcaacctcaa
    13441 ggttaacggc cacgatcccc tcgttgttac tcttggtcag cccattgtcg gtaacgctgg
    13501 ctttgctaac tgggtcgata aactcttctt tggccaggag aaccccgatg tctccaaggt
    13561 gtccaaagac cgaaagctct accgaatcac ccaccgagga gatatcgtcc ctcaagtgcc
    13621 cttctgggac ggttaccagc actgctctgg tgaggtcttt attgactggc ccctgatcca
    13681 ccctcctctc tccaacgttg tcatgtgcca gggccagagc aataaacagt gctctgccgg
    13741 taacactctg ctccagcagg tcaatgtgat tggaaaccat ctgcagtact tcgtcaccga
    13801 gggtgtctgt ggtatctaag ctatttatca ctctttacaa cttctacctc aactatctac
    13861 tttaataaat gaatatcgtt tattctctat gattactgta tatgcgttcc tctaagacaa
    13921 atcgaaacca gcatgcgatc gaatggcata caaaagtttc ttccgaagtt gatcaatgtc
    13981 ctgatagtca ggcagcttga gaagattgac acaggtggag gccgtaggga accgatcaac
    14041 ctgtctacca gcgttacgaa tggcaaatga cgggttcaaa gccttgaatc cttgcaatgg
    14101 tgccttggat actgatgtca caaacttaag aagcagccgc ttgtcctctt cctcgaaact
    14161 ctcaaacaca gtccagaagt cctttatagt ttgatctgta tccagatagc ctccgtaatt
    14221 ggtgtgtgtc ttcaaatccc agacgtccac attggcatgt cctccactga taagcatttg
    14281 aagttcatct gcgttgaaca ttgagaccca cgaagggtca atgagctggt atagaccgcc
    14341 caagaatgca tctgataact tcgtataatg tatgctatac gaagttatgt gatatcagat
    14401 ccactagtgg ccttttcaaa aagcggcggt tcgtggttcg tgtttcgtgt tcgtgtcgcg
    14461 atgtatagtt ataattggag gttgatgaat gatgcagcta aatttgggtt tgtggggagt
    14521 gaaattaaaa agctgaagag aagacagacg acgatgacga tggtggaaga atacaggaaa
    14581 gggattgtgt agtattgttg gactttgctc tcaaatctga tctgaatttt ccttgaattt
    14641 cctcaataat ctcaatattc acgagtttgc agtaggcggt tcttgacgtg ctcttcccca
    14701 tgtacaaatc caattcatca agatcaaatg cacgcacgga aatcgaactc ccaaccacac
    14761 accgcccatc tgcataaact ctaaatgcgg tgaaatttgt aacttctgaa agacacgatt
    14821 ttacacactc acgcaaccac ctgaaacgcc atgttctaca aagtcacaca ggtccgatct
    14881 atcctgcgaa tgccgtggag aacacgtgac gtgctcaaga ccctcggtct gggcaaggtc
    14941 ggctcccaga agtactacaa ggtgtctcct ggtatcgccg gtcagctcta caaggtcaag
    15001 gagctggtca acgtggagat ggccgaccag tacaagtcca aggcccagat caaggccgac
    15061 cgaaaaaccg agcccggatt ctccgttgtc ggttctaagc gagcctaaga gtgagacaga
    15121 atgatctcaa acgacacaaa acgacaggct gatttgaaac aacacaacac aagtgcgagc
    15181 cgatcttgcg gatctggaca ctctaaacat caagtcgaca gtcgccacac atgtatatat
    15241 ccaatgtatt tatcgagttg aaggaaggaa gagctaggaa aggaagctca gtatgcgctt
    15301 ttgagatcgt gttgatgtcg gcagccggag catccgtggg agagtacgag tactgtcggt
    15361 gtgtacaagc cggcagaccc cgggtagaat ggtacaagta gtgtagtgta gtgtactgta
    15421 ctcgtacttt atgggcctat gttcgagtac ttctttcttg tcatgaatgg tctattgatg
    15481 tatatgtcgt aagatatatg tcgtaagaag gtctcctcgt ctttgggcaa gtgctatgaa
    15541 tcatgtaggg gaaagagaac ttggcatatt tagtgttcat atctaagatg gtaacacgga
    15601 gacttaattc attgatatct gttaaatctt ctacagcatc tacaagtagt tgtacttgta
    15661 gagatagaca agccatctgc tccaacatct ccaatgtgca tatcgtgcaa ctacaccagt
    15721 gacagtagta caagtggtat attcattcca tctcatctat cacttgcacc actcaagcaa
    15781 atgcattcat ttcctacctt accacctgtc gcgtatcgca aaccagcata gcttacctgg
    15841 acggcataga cggtagaacc aacagaaacg gcataaacgg gtcagctgaa attaagggac
    15901 ttgatattga gacagaatga cgaggatttt agttagaagt accgccatct gctgtatctc
    15961 tgtcggtgcg tggtgttcgt tcgattcaca cattcttaac aacacgttca acaggactcc
    16021 aacaaaattg cctgccactc tctcctcttt tagacggtaa atgcacaacc ataacaacag
    16081 actcaacaag acagaacagc aaaacaagta aagagagagc atacaactcc cccccaaact
    16141 atccaaagtg gctaaagttg aacctgaagc acttattcta gccataatac cgggcgctgg
    16201 aaacatataa atagccccca tcctcctcct tctattctac accaagcaac agcctctctt
    16261 cccattcaat cagcaatgca gctcaagaac atcctcctcc tctctctctc cgccgtcgcc
    16321 ctggcccaga acgcctctcc cgagtccgac aacgccatgg aggcccccca ggacatctcc
    16381 aaactcaagc ccaagtgccc tgagcagaag tgccccaagc ccaaatcgcg tg
//