LOCUS       sequence-462304-       11703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast CEN plasmid for galactose-inducible expression of spCas9 and 
            Ec86 reverse transcriptase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11703)
  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 11703)
  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-GAL-Hyg
FEATURES             Location/Qualifiers
     source          1..11703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(116..184,185..976)
                     /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(334..353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1147..1735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1636..1655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1889..1906
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2023..2044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2059..2064,2065..2082,2083..2089)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2097..2113
                     /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     2102..2124
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2121..2137
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2121..2137
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2158..2176
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            2304..3879
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        2304..2647
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2648..3676
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             2648..3676
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      3682..3879
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3995..4014)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3996..4014)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4021..4038)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4021..4037)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4030..4052)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4178..4633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4265..4284)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4475..4496
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5938..5958)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        6180..6621
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6180..6297
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6592..6615
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             6654..10757
                     /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             6654..10757
                     /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             6654..10757
                     /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             10770..10790
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(10822..10840)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      10852..11041
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     11055..11077
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(11115..11133)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    11174..11677
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
       61 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      121 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      181 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      241 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      301 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
      361 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
      421 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
      481 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
      541 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
      601 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
      661 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
      721 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
      781 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
      841 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
      901 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
      961 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1261 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     1321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1741 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1861 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1921 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1981 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     2041 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     2101 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat
     2161 taaccctcac taaagggaac aaaagctgct agagcggccg ccagctgaag cttcgtacgc
     2221 tgcaggtcga cggatccccg ggttaattaa ggcgcgccag atctgtttag cttgcctcgt
     2281 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     2341 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     2401 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     2461 gctcctcgct gcggacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
     2521 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     2581 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     2641 aacaaccatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     2701 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     2761 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     2821 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     2881 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     2941 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     3001 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     3061 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     3121 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     3181 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     3241 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     3301 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     3361 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     3421 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     3481 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     3541 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     3601 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     3661 gagggcaaag gaataatcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     3721 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     3781 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     3841 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     3901 cgccatccag tgtcgaaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc
     3961 tatgcggccg ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt attacaattc
     4021 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4081 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4141 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     4201 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4261 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4321 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4381 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4441 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4501 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4561 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4621 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4681 gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     4741 gcatgcgaat tgcatatctt tccatagatt ttcaattttt gatgtctcca tggtggtaca
     4801 gagaacttgt aaacaattcg gtccctacat gtgaggaaat tcgctgtgac acttttatca
     4861 ctgaactcca aatttaaaaa atagcataaa attcgttata cagcaaatct atgtgttgca
     4921 attaagaact aaaagatata gagtgcatat tttcaagaag gatagtaagc tggcaaatca
     4981 agtcttagcc ttgttcaatg ggttcttacc gtacttcttt tccaacttag agatgtaggt
     5041 aatcaatctt ctgtgggact tagaatcaac ggacaagatg aaagacaacc aacctctaac
     5101 atgttcgatt tcggaggatt taccgcagaa aatatggtgg atcttggctc tgatttcttt
     5161 gtacttttct ctaccaatac caaccttttc ttgggagata accaaaccgg taacttttct
     5221 ttgggatctt ggaccagaaa tacaggtctt tttggagttg atgaccaaac cttctgatgg
     5281 aataatggag aacaagaagt ctctggcctt aacaactttc ttcatagatt gagcagacaa
     5341 ggtcaagtca tcagcgtatc tagtgtatat caaacctcta gaaccagcat aaccttgaat
     5401 tctgtagtcc aacttggagc aaatcaagtt agccaatttt ggagaagatg gagcaccttg
     5461 tggcaacaag ttcttgtagc agcaaatctt ggtcaaaaca gaggagatca atctgttgta
     5521 acccaaagaa tggaaaacac cgaaaacctt gttagcagtc aaagatggga agaaatcttc
     5581 caagtcgatg ttcaaaatga agttggcacc aatatgtgga gtggcattgt tcaagatgga
     5641 ttggtgcttt tcaaaaccga tagaaaatgg ggaggaggat aacttgtcca agatgtttct
     5701 caaaacccaa ccttgtaaag ccttcaattc tctggatggt tggtagatag ttctcattct
     5761 cttttctgga cccttctttt caacagtgta gattctgtat ctgaaatcgg cggtgtagat
     5821 caataatctc aaggtttcaa cagaaattct ggtagcctta gacatatcgt gcaagttgtt
     5881 catgactggc aaacccaaat ttctcaatct gaaggtgttc aagtattcag cggacttgac
     5941 ctttcttttc ttctttggtg gcatttatat tgaattttca aaaattctta cttttttttt
     6001 ggatggacgc aaagaagttt aataatcata ttacatggca ttaccaccat atacatatcc
     6061 atatctaatc ttacttatat gttgtggaaa tgtaaagagc cccattatct tagcctaaaa
     6121 aaaccttctc tttggaactt tcagtaatac gcttaactgc tcattgctat attgaagtac
     6181 ggattagaag ccgccgagcg ggcgacagcc ctccgacgga agactctcct ccgtgcgtcc
     6241 tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac
     6301 aataaagatt ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct
     6361 ggccccacaa accttcaaat taacgaatca aattaacaac cataggatga taatgcgatt
     6421 agttttttag ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta
     6481 acagatatat aaatggaaaa gctgcataac cactttaact aatactttca acattttcag
     6541 tttgtattac ttcttattca aatgtcataa aagtatcaac aaaaaattgt taatatacct
     6601 ctatacttta acgtcaagga gaaaaaacaa ctagtggatc ccccgggaaa aaaatggaca
     6661 agaagtactc cattgggctc gatatcggca caaacagcgt cggttgggcc gtcattacgg
     6721 acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat cgccacagca
     6781 taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc gaagccacgc
     6841 ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc tgctacctgc
     6901 aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat aggctggagg
     6961 agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt ggcaatatcg
     7021 tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg aagaagcttg
     7081 tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg catatgatca
     7141 aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc gatgtcgaca
     7201 aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac ccgatcaacg
     7261 catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc cggcggctcg
     7321 aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt aatcttatcg
     7381 ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc gaagatgcca
     7441 agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg gcccagatcg
     7501 gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc attctgctga
     7561 gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct agtatgatca
     7621 agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc agacagcaac
     7681 tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac gccggataca
     7741 ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc ttggaaaaaa
     7801 tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg cgcaaacagc
     7861 gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg cacgctatcc
     7921 tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag attgagaaaa
     7981 tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat tccagattcg
     8041 cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag gaagtcgtgg
     8101 ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat aaaaatctgc
     8161 ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca gtttataacg
     8221 agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc ctgtctggag
     8281 agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt accgtgaaac
     8341 agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa atcagcggag
     8401 tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa atcattaaag
     8461 acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt gtcctcaccc
     8521 ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac gctcatctct
     8581 tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg gggcggctgt
     8641 caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc ctggattttc
     8701 ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat gactctctca
     8761 cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt cttcacgagc
     8821 acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag accgttaagg
     8881 tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc gttatcgaga
     8941 tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa aggatgaaga
     9001 ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac ccagttgaaa
     9061 acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc agggacatgt
     9121 acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggat catatcgtgc
     9181 cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga tccgataaaa
     9241 atagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg aaaaattatt
     9301 ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat ctgactaagg
     9361 ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg cagcttgttg
     9421 agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg aacaccaagt
     9481 acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag tctaagctgg
     9541 tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac aattaccacc
     9601 atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa aaatatccca
     9661 agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg aaaatgatcg
     9721 caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac agcaatatta
     9781 tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag cgaccactta
     9841 tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag
     9901 tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg
     9961 gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg atcgcacgca
    10021 aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc gcttacagtg
    10081 tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac
    10141 tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg
    10201 aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc aagtactctc
    10261 tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag
    10321 gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc agccactatg
    10381 aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg gaacaacaca
    10441 aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg
    10501 ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat aagcccatca
    10561 gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc gcgcctgcag
    10621 ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca aaggaggtcc
    10681 tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga atcgacctct
    10741 ctcagctcgg tggagacagc agggctgacc ccaagaagaa gaggaaggtg tgatctcttc
    10801 tcgagtcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
    10861 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
    10921 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
    10981 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
    11041 ggctttaatt tcatccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
    11101 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
    11161 ggtttcttag acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg
    11221 aataatttgg gaatttactc tgtgtttatt tatttttatg ttttgtattt ggattttaga
    11281 aagtaaataa agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta
    11341 aaaaatttca acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta
    11401 aatagatata cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg
    11461 gtcttctaca cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa
    11521 gataaaaggt agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa
    11581 aactattttt tctttaattt ctttttttac tttctatttt taatttatat atttatatta
    11641 aaaaatttaa attataatta tttttatagc acgtgatgaa aaggacccag gtggcacttt
    11701 tcg
//
