LOCUS       sequence-478303-        9833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9-carrying vector with kanMX marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9833)
  AUTHORS   Stovicek V, Borodina I, Forster J
  TITLE     CRISPR-Cas system enables fast and simple genome editing of 
            industrial Saccharomyces cerevisiae strains.
  JOURNAL   Metab Eng Commun. 2015 Mar 20;2:13-22. doi: 
            10.1016/j.meteno.2015.03.001. eCollection 2015 Dec.
  PUBMED    34150504
REFERENCE   2  (bases 1 to 9833)
  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 
            Commun."; date: "2015-03-20"; volume: "2:13-22. doi"; pages: " 
            10.1016/j.meteno.2015.03.001. eCollection 2015 Dec"
COMMENT     Sequence Label: pCfB2312 (TEF1p-Cas9-CYC1t_kanMX)
FEATURES             Location/Qualifiers
     source          1..9833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..504
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        541..645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(646..714,715..1506)
                     /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(864..883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1677..2265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2166..2185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2419..2436
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2553..2574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2589..2594,2595..2612,2613..2619)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2627..2643
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2632..2654
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2651..2667
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2651..2667
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2686..2706
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2688..2706
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2724..3122
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3126..3142
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3126..3142
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3154..7257
                     /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             3154..7257
                     /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             3154..7257
                     /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             7270..7290
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7306..7553
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7322..7340)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7571..7590)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7572..7590)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7600..7617)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7600..7616)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7609..7631)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7757..8137
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7844..7863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8054..8075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    complement(8189..8222)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            complement(8285..9641)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(8285..8482)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(8488..9297)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     9211..9230
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(9298..9641)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    complement(9696..9729)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa
       61 tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga
      121 aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca
      181 aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat agatatacat
      241 tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag
      301 acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt
      361 atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct
      421 ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt
      481 ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg
      541 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      601 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      661 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      721 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      781 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      841 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      901 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      961 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1021 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1081 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1141 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1201 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1261 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1321 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1381 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1441 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1501 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1561 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1621 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1681 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1741 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1801 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1861 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1921 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1981 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2041 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2101 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2161 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2221 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2281 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2341 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2401 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2461 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2521 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     2581 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2641 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     2701 taaagggaac aaaagctgga gctcatagct tcaaaatgtt tctactcctt ttttactctt
     2761 ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata
     2821 ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga
     2881 aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt
     2941 atcacgtttc tttttcttga aaattttttt tttgattttt ttctctttcg atgacctccc
     3001 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
     3061 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
     3121 agttttctag aactagtgga tcccccggga aaaatggaca agaagtactc cattgggctc
     3181 gatatcggca caaacagcgt cggttgggcc gtcattacgg acgagtacaa ggtgccgagc
     3241 aaaaaattca aagttctggg caataccgat cgccacagca taaagaagaa cctcattggc
     3301 gccctcctgt tcgactccgg ggagacggcc gaagccacgc ggctcaaaag aacagcacgg
     3361 cgcagatata cccgcagaaa gaatcggatc tgctacctgc aggagatctt tagtaatgag
     3421 atggctaagg tggatgactc tttcttccat aggctggagg agtccttttt ggtggaggag
     3481 gataaaaagc acgagcgcca cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat
     3541 gaaaagtacc caaccatata tcatctgagg aagaagcttg tagacagtac tgataaggct
     3601 gacttgcggt tgatctatct cgcgctggcg catatgatca aatttcgggg acacttcctc
     3661 atcgaggggg acctgaaccc agacaacagc gatgtcgaca aactctttat ccaactggtt
     3721 cagacttaca atcagctttt cgaagagaac ccgatcaacg catccggagt tgacgccaaa
     3781 gcaatcctga gcgctaggct gtccaaatcc cggcggctcg aaaacctcat cgcacagctc
     3841 cctggggaga agaagaacgg cctgtttggt aatcttatcg ccctgtcact cgggctgacc
     3901 cccaacttta aatctaactt cgacctggcc gaagatgcca agcttcaact gagcaaagac
     3961 acctacgatg atgatctcga caatctgctg gcccagatcg gcgaccagta cgcagacctt
     4021 tttttggcgg caaagaacct gtcagacgcc attctgctga gtgatattct gcgagtgaac
     4081 acggagatca ccaaagctcc gctgagcgct agtatgatca agcgctatga tgagcaccac
     4141 caagacttga ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa
     4201 attttcttcg atcagtctaa aaatggctac gccggataca ttgacggcgg agcaagccag
     4261 gaggaatttt acaaatttat taagcccatc ttggaaaaaa tggacggcac cgaggagctg
     4321 ctggtaaagc ttaacagaga agatctgttg cgcaaacagc gcactttcga caatggaagc
     4381 atcccccacc agattcacct gggcgaactg cacgctatcc tcaggcggca agaggatttc
     4441 tacccctttt tgaaagataa cagggaaaag attgagaaaa tcctcacatt tcggataccc
     4501 tactatgtag gccccctcgc ccggggaaat tccagattcg cgtggatgac tcgcaaatca
     4561 gaagagacca tcactccctg gaacttcgag gaagtcgtgg ataagggggc ctctgcccag
     4621 tccttcatcg aaaggatgac taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct
     4681 aaacactctc tgctgtacga gtacttcaca gtttataacg agctcaccaa ggtcaaatac
     4741 gtcacagaag ggatgagaaa gccagcattc ctgtctggag agcagaagaa agctatcgtg
     4801 gacctcctct tcaagacgaa ccggaaagtt accgtgaaac agctcaaaga agactatttc
     4861 aaaaagattg aatgtttcga ctctgttgaa atcagcggag tggaggatcg cttcaacgca
     4921 tccctgggaa cgtatcacga tctcctgaaa atcattaaag acaaggactt cctggacaat
     4981 gaggagaacg aggacattct tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg
     5041 gagatgattg aagaacgctt gaaaacttac gctcatctct tcgacgacaa agtcatgaaa
     5101 cagctcaaga ggcgccgata tacaggatgg gggcggctgt caagaaaact gatcaatggg
     5161 atccgagaca agcagagtgg aaagacaatc ctggattttc ttaagtccga tggatttgcc
     5221 aaccggaact tcatgcagtt gatccatgat gactctctca cctttaagga ggacatccag
     5281 aaagcacaag tttctggcca gggggacagt cttcacgagc acatcgctaa tcttgcaggt
     5341 agcccagcta tcaaaaaggg aatactgcag accgttaagg tcgtggatga actcgtcaaa
     5401 gtaatgggaa gacataagcc cgagaatatc gttatcgaga tggcccgaga gaaccaaact
     5461 acccagaagg gacagaagaa cagtagggaa aggatgaaga ggattgaaga gggtataaaa
     5521 gaactggggt cccaaatcct taaggaacac ccagttgaaa acacccagct tcagaatgag
     5581 aagctctacc tgtactacct gcagaacggc agggacatgt acgtggatca ggaactggac
     5641 atcaatcggc tctccgacta cgacgtggat catatcgtgc cccagtcttt tctcaaagat
     5701 gattctattg ataataaagt gttgacaaga tccgataaaa atagagggaa gagtgataac
     5761 gtcccctcag aagaagttgt caagaaaatg aaaaattatt ggcggcagct gctgaacgcc
     5821 aaactgatca cacaacggaa gttcgataat ctgactaagg ctgaacgagg tggcctgtct
     5881 gagttggata aagccggctt catcaaaagg cagcttgttg agacacgcca gatcaccaag
     5941 cacgtggccc aaattctcga ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg
     6001 attcgagagg tgaaagttat tactctgaag tctaagctgg tctcagattt cagaaaggac
     6061 tttcagtttt ataaggtgag agagatcaac aattaccacc atgcgcatga tgcctacctg
     6121 aatgcagtgg taggcactgc acttatcaaa aaatatccca agcttgaatc tgaatttgtt
     6181 tacggagact ataaagtgta cgatgttagg aaaatgatcg caaagtctga gcaggaaata
     6241 ggcaaggcca ccgctaagta cttcttttac agcaatatta tgaatttttt caagaccgag
     6301 attacactgg ccaatggaga gattcggaag cgaccactta tcgaaacaaa cggagaaaca
     6361 ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag tccggaaggt cctgtccatg
     6421 ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg gaggcttctc caaggaaagt
     6481 atcctcccga aaaggaacag cgacaagctg atcgcacgca aaaaagattg ggaccccaag
     6541 aaatacggcg gattcgattc tcctacagtc gcttacagtg tactggttgt ggccaaagtg
     6601 gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac tgctgggcat cacaatcatg
     6661 gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg aggcgaaagg atataaagag
     6721 gtcaaaaaag acctcatcat taagcttccc aagtactctc tctttgagct tgaaaacggc
     6781 cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc
     6841 tctaaatacg ttaatttctt gtatctggcc agccactatg aaaagctcaa agggtctccc
     6901 gaagataatg agcagaagca gctgttcgtg gaacaacaca aacactacct tgatgagatc
     6961 atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag
     7021 gtgctttctg cttacaataa gcacagggat aagcccatca gggagcaggc agaaaacatt
     7081 atccacttgt ttactctgac caacttgggc gcgcctgcag ccttcaagta cttcgacacc
     7141 accatagaca gaaagcggta cacctctaca aaggaggtcc tggacgccac actgattcat
     7201 cagtcaatta cggggctcta tgaaacaaga atcgacctct ctcagctcgg tggagacagc
     7261 agggctgacc ccaagaagaa gaggaaggtg tgatctcttc tcgagtcatg taattagtta
     7321 tgtcacgctt acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta
     7381 gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt
     7441 tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta
     7501 tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcggccggt
     7561 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     7621 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     7681 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     7741 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     7801 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     7861 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     7921 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     7981 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     8041 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     8101 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa ttgggtgcat
     8161 aggccactag tggatctgat atcacctaat aacttcgtat agcatacatt atacgaagtt
     8221 atattaaggg ttctcgagag ctcgttttcg acactggatg gcggcgttag tatcgaatcg
     8281 acagcagtat agcgaccagc attcacatac gattgacgca tgatattact ttctgcgcac
     8341 ttaacttcgc atctgggcag atgatgtcga ggcgaaaaaa aatataaatc acgctaacat
     8401 ttgattaaaa tagaacaact acaatataaa aaaactatac aaatgacaag ttcttgaaaa
     8461 caagaatctt tttattgtca gtactgatta gaaaaactca tcgagcatca aatgaaactg
     8521 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     8581 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     8641 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     8701 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     8761 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     8821 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     8881 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     8941 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttgccggg
     9001 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     9061 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     9121 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     9181 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     9241 agcatccatg ttggaattta atcgcggcct cgaaacgtga gtcttttcct tacccatggt
     9301 tgtttatgtt cggatgtgat gtgagaactg tatcctagca agattttaaa aggaagtata
     9361 tgaaagaaga acctcagtgg caaatcctaa ccttttatat ttctctacag gggcgcggcg
     9421 tggggacaat tcaacgcgtc tgtgagggga gcgtttccct gctcgcaggt ctgcagcgag
     9481 gagccgtaat ttttgcttcg cgccgtgcgg ccatcaaaat gtatggatgc aaatgattat
     9541 acatggggat gtatgggcta aatgtacggg cgacagtcac atcatgcccc tgagctgcgc
     9601 acgtcaagac tgtcaaggag ggtattctgg gcctccatgt cgctggccgg gtgacccggc
     9661 ggggacgagg caagctaaac agatctctag acctaataac ttcgtatagc atacattata
     9721 cgaagttata ttaagggttg tcgacctgca gcgtacgaag cttcagctga cgcgatctcg
     9781 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acg
//
