LOCUS       sequence-462039-       13320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast CEN plasmid for estradiol-inducible expression of spCas9 and 
            Ec86 reverse transcriptase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13320)
  AUTHORS   Stuecker TN, Hood SE, Molina Pineda J, Lenaduwe S, Winter J, Sadhu 
            MJ, Lewis JA
  TITLE     Improved vectors for retron-mediated CRISPR-Cas9 genome editing in 
            Saccharomyces cerevisiae.
  JOURNAL   G3 (Bethesda). 2025 Aug 4:jkaf175. doi: 10.1093/g3journal/jkaf175.
  PUBMED    40758833
REFERENCE   2  (bases 1 to 13320)
  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: "G3 
            (Bethesda). 2025 Aug 4:jkaf175. doi: 10.1093/g3journal/jkaf175."
COMMENT     Sequence Label: pCas9-EcRT-Z3-Nat
FEATURES             Location/Qualifiers
     source          1..13320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     79..96
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    213..234
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(249..254,255..272,273..279)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    287..303
                     /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     292..314
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     311..327
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     311..327
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        348..366
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            494..1613
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        494..837
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             838..1410
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNALAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1416..1613
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1729..1748)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1730..1748)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1755..1772)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1755..1771)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1764..1786)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1912..2367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1999..2018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2209..2230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5699..5719)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     6395..6418
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             6461..10564
                     /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             6461..10564
                     /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             6461..10564
                     /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             10577..10597
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(10629..10647)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      10659..10848
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     10862..10884
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10922..10940)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    10981..11484
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        11521..11625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11626..11694,11695..12486)
                     /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(11844..11863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12657..13245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13146..13165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
       61 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
      121 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
      181 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      241 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      301 caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac
      361 taaagggaac aaaagctgct agagcggccg ccagctgaag cttcgtacgc tgcaggtcga
      421 cggatccccg ggttaattaa ggcgcgccag atctgtttag cttgcctcgt ccccgccggg
      481 tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag
      541 ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc
      601 atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct
      661 gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc
      721 gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact
      781 tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg
      841 ggtaccactc ttgacgacac ggcttaccgg taccgcacca gtgtcccggg ggacgccgag
      901 gccatcgagg cactggatgg gtccttcacc accgacaccg tcttccgcgt caccgccacc
      961 ggggacggct tcaccctgcg ggaggtgccg gtggacccgc ccctgaccaa ggtgttcccc
     1021 gacgacgaat cggacgacga atcggacgac ggggaggacg gcgacccgga ctcccggacg
     1081 ttcgtcgcgt acggggacga cggcgacctg gcgggcttcg tggtcgtctc gtactccggc
     1141 tggaaccgcc ggctgaccgt cgaggacatc gaggtcgccc cggagcaccg ggggcacggg
     1201 gtcgggcgcg cgttgatggg gctcgcgacg gagttcgccc gcgagcgggg cgccgggcac
     1261 ctctggctgg aggtcaccaa cgtcaacgca ctggcgatcc acgcgtaccg gcggatgggg
     1321 ttcaccctct gcggcctgga caccgccctg tacgacggca ccgcctcgga cggcgagcag
     1381 gcgctctaca tgagcatgcc ctgcccctaa tcagtactga caataaaaag attcttgttt
     1441 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     1501 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     1561 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     1621 ttcgatacta acgccgccat ccagtgtcga aaacgagctc gaattcatcg atgatatcag
     1681 atccactagt ggcctatgcg gccgccaccg cggtggagct ccaattcgcc ctatagtgag
     1741 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1801 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1861 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     1921 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1981 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2041 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2101 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2161 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2221 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2281 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2341 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     2401 cggtatttca caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg
     2461 agtttagtat acatgcatgc gtcgatctcc cctcaagtgg caaactgcga ctgatatgaa
     2521 agtaaagtat cggaccttct attacggaat gtatacaaac gctacaggat acagtgatat
     2581 taaatagaaa acaagtctac aattaaatgg cagaaagaag aaccagctta ctactaacca
     2641 ccatactcat caatacccag agcatcggtg aacatctgct caaattcgaa gtcagccata
     2701 tccagcgcac cgtaaggtgc tgaatcatgt ggagtgaagc caggccctgg agagtccccg
     2761 tcacccaaca tatctaaatc aaaatcgtct aaagcgtcag cgtgggccat ggcaacgtct
     2821 tcaccgtcca aatgcagttc gctagatgta gaccctgctg tcgctaggtg ggattggtcg
     2881 gtttcttcaa cggaggctcc gcctctagaa gttggtgcat gcaacctgtg agcgtccagc
     2941 atttctagta ataaatcata cagaggaacg acgtttttac acttcataga atacaggtgc
     3001 tccataccct tgttagacat atgtctaatg tgtgacaaaa ttagtagtaa ctgagctagc
     3061 ctctgatgct gttgctgcag agtcaatcct gctttcgcca taaggtgaat caatgtatca
     3121 gtgatcttgt ccaacactct atgtatatgg tctttctctt ccaggctttt cagagttgag
     3181 cttaaaaatg tatagacacc gctatttaac aaaataattg atttcaggca aacgaattct
     3241 tctccttgca aattcatcat tctgaacctg gagctagtgg caagaagcat atcaaaaatc
     3301 tcaaccatac cttctacgca tttaccttgg ttcctgtcca ataataaatt aggtgcaaat
     3361 aataatttta cagggtgctc catacttctc cacactaatc cgatcatcaa tatttctaac
     3421 cacgcgcatt caagaagatg tacctggtca tgtaatgtca agtcaacaaa gccgggaact
     3481 cttttggccc aattaatcat atgaactaat tccctgtctg ctagatttgt caaaagaccc
     3541 atcatggaag cctcggaaaa aggacgagtt gggtcgtatt cactgtacaa tataggtggt
     3601 tcagcatcaa gtagtgcgga aaccatttga tcagcggtca aagacagtgc aagagaattc
     3661 ttcttagatc ttttgatcat taatgggctt ggccacaagt tggctgccct catgtctcca
     3721 gcgcttggat cttctagggt tgaggctgct gccggagtgc ccccacctcc tgtatggatc
     3781 tttgtatgtc tctttctttc atcagatctg gcgaactttc ttccacaaat gtcacacgcg
     3841 aacggcttct cgccggtgtg ggttctaatg tgcgtagtca aatggtcact tctgctgaag
     3901 tttctcatac agattctaca ttggaaaggc ttttggccgg tgtgaatacg aatatgccta
     3961 gtcagctcgt cagatctgct gaatctccta tcacaacttt ctacagggca agcgtagggt
     4021 ctcgttccca ttgttaattc agtaaatttt cgatcttggg aagaaaaaag cagtaagcgt
     4081 gaaaaatcta aaagctgatg tagtagaaga tcctattctt taacaaagat tgaccttttc
     4141 tttttcttct tggtttgagt agaaagggga aggaagaata caagagagag gaaaaaaagg
     4201 aagataaaaa gagagcgtga tataaatgaa tatatattaa acaagagaga ttgggaagga
     4261 aaggatcaaa caaacccaaa aatatttcaa aaaggagaga gagaggcgag tttggtttca
     4321 aaacggttta tttatttatg caagaggacg tggaagaaaa agaagaagga agaaaaaaat
     4381 ttgaaagaaa aaaacgcgtg gcgggtaaag aagaaaatgg aaaatagagg ccgggtgaca
     4441 gagaaatatt gagggttaat tggaaaatat gttagggtga ggcatatgtt tttaaaggga
     4501 tgcatgcgaa ttgcatatct ttccatagat tttcaatttt tgatgtctcc atggtggtac
     4561 agagaacttg taaacaattc ggtccctaca tgtgaggaaa ttcgctgtga cacttttatc
     4621 actgaactcc aaatttaaaa aatagcataa aattcgttat acagcaaatc tatgtgttgc
     4681 aattaagaac taaaagatat agagtgcata ttttcaagaa ggatagtaag ctggcaaatc
     4741 aagtcttagc cttgttcaat gggttcttac cgtacttctt ttccaactta gagatgtagg
     4801 taatcaatct tctgtgggac ttagaatcaa cggacaagat gaaagacaac caacctctaa
     4861 catgttcgat ttcggaggat ttaccgcaga aaatatggtg gatcttggct ctgatttctt
     4921 tgtacttttc tctaccaata ccaacctttt cttgggagat aaccaaaccg gtaacttttc
     4981 tttgggatct tggaccagaa atacaggtct ttttggagtt gatgaccaaa ccttctgatg
     5041 gaataatgga gaacaagaag tctctggcct taacaacttt cttcatagat tgagcagaca
     5101 aggtcaagtc atcagcgtat ctagtgtata tcaaacctct agaaccagca taaccttgaa
     5161 ttctgtagtc caacttggag caaatcaagt tagccaattt tggagaagat ggagcacctt
     5221 gtggcaacaa gttcttgtag cagcaaatct tggtcaaaac agaggagatc aatctgttgt
     5281 aacccaaaga atggaaaaca ccgaaaacct tgttagcagt caaagatggg aagaaatctt
     5341 ccaagtcgat gttcaaaatg aagttggcac caatatgtgg agtggcattg ttcaagatgg
     5401 attggtgctt ttcaaaaccg atagaaaatg gggaggagga taacttgtcc aagatgtttc
     5461 tcaaaaccca accttgtaaa gccttcaatt ctctggatgg ttggtagata gttctcattc
     5521 tcttttctgg acccttcttt tcaacagtgt agattctgta tctgaaatcg gcggtgtaga
     5581 tcaataatct caaggtttca acagaaattc tggtagcctt agacatatcg tgcaagttgt
     5641 tcatgactgg caaacccaaa tttctcaatc tgaaggtgtt caagtattca gcggacttga
     5701 cctttctttt cttctttggt ggcatttata ttgaatttat attgaatttt caaaaattct
     5761 tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc
     5821 atatacatat ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa
     5881 gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa
     5941 ctgctcattg ctatattgaa gtgcggccgc gtgggcgtgc gtgggcgggc gtgggcgtgc
     6001 gtgggcgggc gtgggcgtgc gtgggcgtct agaccgtgcg tcctcgtctt caccggtcgc
     6061 gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg aacaataaag attctacaat
     6121 actagctttt atggttatga agaggaaaaa ttggcagtaa cctggcccca caaaccttca
     6181 aattaacgaa tcaaattaac aaccatagga tgataatgcg attagttttt tagccttatt
     6241 tctggggtaa ttaatcagcg aagcgatgat ttttgatcta ttaacagata tataaatgga
     6301 aaagctgcat aaccacttta actaatactt tcaacatttt cagtttgtat tacttcttat
     6361 tcaaatgtca taaaagtatc aacaaaaaat tgttaatata cctctatact ttaacgtcaa
     6421 ggagaaaaaa ctataaacta gtggatcccc cgggaaaaaa atggacaaga agtactccat
     6481 tgggctcgat atcggcacaa acagcgtcgg ttgggccgtc attacggacg agtacaaggt
     6541 gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct
     6601 cattggcgcc ctcctgttcg actccgggga gacggccgaa gccacgcggc tcaaaagaac
     6661 agcacggcgc agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag
     6721 taatgagatg gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt
     6781 ggaggaggat aaaaagcacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc
     6841 gtaccatgaa aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga
     6901 taaggctgac ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca
     6961 cttcctcatc gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca
     7021 actggttcag acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga
     7081 cgccaaagca atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc
     7141 acagctccct ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg
     7201 gctgaccccc aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag
     7261 caaagacacc tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc
     7321 agaccttttt ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg
     7381 agtgaacacg gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga
     7441 gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta
     7501 caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc
     7561 aagccaggag gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga
     7621 ggagctgctg gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa
     7681 tggaagcatc ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga
     7741 ggatttctac ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg
     7801 gataccctac tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg
     7861 caaatcagaa gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc
     7921 tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt
     7981 gcttcctaaa cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt
     8041 caaatacgtc acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc
     8101 tatcgtggac ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga
     8161 ctatttcaaa aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt
     8221 caacgcatcc ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct
     8281 ggacaatgag gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga
     8341 agatagggag atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt
     8401 catgaaacag ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat
     8461 caatgggatc cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg
     8521 atttgccaac cggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga
     8581 catccagaaa gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct
     8641 tgcaggtagc ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact
     8701 cgtcaaagta atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa
     8761 ccaaactacc cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg
     8821 tataaaagaa ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca
     8881 gaatgagaag ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga
     8941 actggacatc aatcggctct ccgactacga cgtggatcat atcgtgcccc agtcttttct
     9001 caaagatgat tctattgata ataaagtgtt gacaagatcc gataaaaata gagggaagag
     9061 tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct
     9121 gaacgccaaa ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg
     9181 cctgtctgag ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat
     9241 caccaagcac gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga
     9301 caaactgatt cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag
     9361 aaaggacttt cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc
     9421 ctacctgaat gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga
     9481 atttgtttac ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca
     9541 ggaaataggc aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa
     9601 gaccgagatt acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg
     9661 agaaacagga gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct
     9721 gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa
     9781 ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga
     9841 ccccaagaaa tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc
     9901 caaagtggag aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac
     9961 aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata
    10021 taaagaggtc aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga
    10081 aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc
    10141 actgccctct aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg
    10201 gtctcccgaa gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga
    10261 tgagatcatc gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct
    10321 cgataaggtg ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga
    10381 aaacattatc cacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt
    10441 cgacaccacc atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact
    10501 gattcatcag tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg
    10561 agacagcagg gctgacccca agaagaagag gaaggtgtga tctcttctcg agtcatgtaa
    10621 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
    10681 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
    10741 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
    10801 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttca
    10861 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
    10921 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
    10981 gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa
    11041 tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga
    11101 aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca
    11161 aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat agatatacat
    11221 tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag
    11281 acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt
    11341 atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct
    11401 ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt
    11461 ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg
    11521 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
    11581 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
    11641 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
    11701 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
    11761 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
    11821 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
    11881 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
    11941 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
    12001 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
    12061 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
    12121 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
    12181 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
    12241 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    12301 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    12361 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    12421 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    12481 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    12541 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    12601 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    12661 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
    12721 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
    12781 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
    12841 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
    12901 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
    12961 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    13021 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    13081 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    13141 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    13201 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    13261 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
//
