LOCUS       sequence-462313-       11265 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 11265)
  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 11265)
  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-HIS3
FEATURES             Location/Qualifiers
     source          1..11265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      773..1361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1262..1281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1515..1532
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1649..1670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1685..1690,1691..1708,1709..1715)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1723..1739
                     /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     1728..1750
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1747..1763
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1747..1763
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1784..1802
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            1930..3130
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1930..2273
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2274..2927
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2933..3130
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3183..3202)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3184..3202)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3209..3226)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3209..3225)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3218..3240)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3366..3821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3453..3472)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3663..3684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5126..5146)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        5368..5809
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    5368..5485
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5780..5803
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             5842..9945
                     /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             5842..9945
                     /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             5842..9945
                     /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             9958..9978
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(10010..10028)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      10040..10229
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     10243..10265
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10303..10321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    10362..10865
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10902..11006
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11007..11075,11076..11265,1..602)
                     /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(11225..11244)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
       61 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      121 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
      181 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
      241 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
      301 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
      361 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
      421 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
      481 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      541 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      601 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      661 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      721 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      781 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      841 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      901 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      961 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1021 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1081 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1141 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1201 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1261 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1321 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1381 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1441 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1501 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     1561 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1621 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1681 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1741 ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa
     1801 gggaacaaaa gctgctagag cggccgccag ctgaagcttc gtacgctgca ggtcgacgga
     1861 tccccgggtt aattaaggcg cgccagatct gtttagcttg cctcgtcccc gccgggtcac
     1921 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
     1981 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
     2041 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
     2101 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
     2161 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
     2221 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatgggta
     2281 ggagggcttt tgtagaaaga aatacgaacg aaacgaaaat cagcgttgcc atcgctttgg
     2341 acaaagctcc cttacctgaa gagtcgaatt ttattgatga acttataact tccaagcatg
     2401 caaaccaaaa gggagaacaa gtaatccaag tagacacggg aattggattc ctggatcaca
     2461 tgtatcatgc actggctaaa catgcaggct ggagcttacg actttactca agaggtgatt
     2521 taatcatcga tgatcatcac actgcagaag atactgctat tgcacttggt attgcattca
     2581 agcaggctat gggtaacttt gccggcgtta aaagatttgg acatgcttat tgtccacttg
     2641 acgaagccct ttctagaagc gtagttgact tgtcgggacg gccctatgct gttatcgatt
     2701 tgggattaaa gcgtgaaaag gttggggaat tgtcctgtga aatgatccct cacttactat
     2761 attccttttc ggtagcagct ggaattactt tgcatgttac ctgcttatat ggtagtaatg
     2821 accatcatcg tgctgaaagc gcttttaaat ctctggctgt tgccatgcgc gcggctacta
     2881 gtcttactgg aagttctgaa gtcccaagca cgaagggagt gttgtaaaga gtactgacaa
     2941 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     3001 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     3061 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     3121 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttcggtgg agctccaatt
     3181 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     3241 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3301 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3361 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3421 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3481 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3541 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3601 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3661 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3721 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3781 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     3841 cttacgcatc tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga
     3901 agctctaatt tgtgagttta gtatacatgc atgcgaattg catatctttc catagatttt
     3961 caatttttga tgtctccatg gtggtacaga gaacttgtaa acaattcggt ccctacatgt
     4021 gaggaaattc gctgtgacac ttttatcact gaactccaaa tttaaaaaat agcataaaat
     4081 tcgttataca gcaaatctat gtgttgcaat taagaactaa aagatataga gtgcatattt
     4141 tcaagaagga tagtaagctg gcaaatcaag tcttagcctt gttcaatggg ttcttaccgt
     4201 acttcttttc caacttagag atgtaggtaa tcaatcttct gtgggactta gaatcaacgg
     4261 acaagatgaa agacaaccaa cctctaacat gttcgatttc ggaggattta ccgcagaaaa
     4321 tatggtggat cttggctctg atttctttgt acttttctct accaatacca accttttctt
     4381 gggagataac caaaccggta acttttcttt gggatcttgg accagaaata caggtctttt
     4441 tggagttgat gaccaaacct tctgatggaa taatggagaa caagaagtct ctggccttaa
     4501 caactttctt catagattga gcagacaagg tcaagtcatc agcgtatcta gtgtatatca
     4561 aacctctaga accagcataa ccttgaattc tgtagtccaa cttggagcaa atcaagttag
     4621 ccaattttgg agaagatgga gcaccttgtg gcaacaagtt cttgtagcag caaatcttgg
     4681 tcaaaacaga ggagatcaat ctgttgtaac ccaaagaatg gaaaacaccg aaaaccttgt
     4741 tagcagtcaa agatgggaag aaatcttcca agtcgatgtt caaaatgaag ttggcaccaa
     4801 tatgtggagt ggcattgttc aagatggatt ggtgcttttc aaaaccgata gaaaatgggg
     4861 aggaggataa cttgtccaag atgtttctca aaacccaacc ttgtaaagcc ttcaattctc
     4921 tggatggttg gtagatagtt ctcattctct tttctggacc cttcttttca acagtgtaga
     4981 ttctgtatct gaaatcggcg gtgtagatca ataatctcaa ggtttcaaca gaaattctgg
     5041 tagccttaga catatcgtgc aagttgttca tgactggcaa acccaaattt ctcaatctga
     5101 aggtgttcaa gtattcagcg gacttgacct ttcttttctt ctttggtggc atttatattg
     5161 aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa taatcatatt
     5221 acatggcatt accaccatat acatatccat atctaatctt acttatatgt tgtggaaatg
     5281 taaagagccc cattatctta gcctaaaaaa accttctctt tggaactttc agtaatacgc
     5341 ttaactgctc attgctatat tgaagtacgg attagaagcc gccgagcggg cgacagccct
     5401 ccgacggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag
     5461 atgtgcctcg cgccgcactg ctccgaacaa taaagattct acaatactag cttttatggt
     5521 tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatta acgaatcaaa
     5581 ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg ggtaattaat
     5641 cagcgaagcg atgatttttg atctattaac agatatataa atggaaaagc tgcataacca
     5701 ctttaactaa tactttcaac attttcagtt tgtattactt cttattcaaa tgtcataaaa
     5761 gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga aaaaacaact
     5821 agtggatccc ccgggaaaaa aatggacaag aagtactcca ttgggctcga tatcggcaca
     5881 aacagcgtcg gttgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa
     5941 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc
     6001 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc
     6061 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg
     6121 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac
     6181 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca
     6241 accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg
     6301 atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac
     6361 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat
     6421 cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc
     6481 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag
     6541 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa
     6601 tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat
     6661 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca
     6721 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc
     6781 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact
     6841 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat
     6901 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac
     6961 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt
     7021 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag
     7081 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg
     7141 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc
     7201 cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc
     7261 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa
     7321 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg
     7381 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg
     7441 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc
     7501 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa
     7561 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg
     7621 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag
     7681 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa
     7741 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg
     7801 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag
     7861 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc
     7921 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt
     7981 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc
     8041 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg
     8101 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga
     8161 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc
     8221 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg
     8281 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc
     8341 tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga ttctattgat
     8401 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa
     8461 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca
     8521 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa
     8581 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa
     8641 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg
     8701 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat
     8761 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta
     8821 ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat
     8881 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc
     8941 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc
     9001 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg
     9061 tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac
     9121 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa
     9181 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga
     9241 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag
     9301 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc
     9361 ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac
     9421 ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg
     9481 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt
     9541 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag
     9601 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata
     9661 agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct
     9721 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt
     9781 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga
     9841 aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg
     9901 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacagcag ggctgacccc
     9961 aagaagaaga ggaaggtgtg atctcttctc gagtcatgta attagttatg tcacgcttac
    10021 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
    10081 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
    10141 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
    10201 tgcttgagaa ggttttggga cgctcgaagg ctttaatttc atccgggagc tgcatgtgtc
    10261 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
    10321 ttttataggt taatgtcatg ataataatgg tttcttagac ggatcgcttg cctgtaactt
    10381 acacgcgcct cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta
    10441 tttttatgtt ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa
    10501 tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat
    10561 atatttatta gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa
    10621 atgtaaaatc acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg
    10681 cattaatacc tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct
    10741 agagtctttt acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt
    10801 tctattttta atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac
    10861 gtgatgaaaa ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
    10921 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
    10981 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
    11041 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
    11101 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
    11161 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
    11221 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaag
//
