LOCUS       sequence-435228-       11323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating inducible cassette (Cas9-P2A-Csy4 and Eco1 RT) for yeast
            overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11323)
  AUTHORS   Gonzalez-Delgado A, Lopez SC, Rojas-Montero M, Fishman CB, Shipman 
            SL
  TITLE     Simultaneous multi-site editing of individual genomes using retron 
            arrays.
  JOURNAL   Nat Chem Biol. 2024 Nov;20(11):1482-1492. doi: 
            10.1038/s41589-024-01665-7. Epub 2024 Jul 9.
  PUBMED    38982310
REFERENCE   2  (bases 1 to 11323)
  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"; doi: 
            "10.1038/s41589-024-01665-7"; journalName: "Nat Chem Biol"; date: 
            "2024-11"; volume: "20"; issue: "11"; pages: "1482-1492"
COMMENT     Sequence Label: pSCL390
FEATURES             Location/Qualifiers
     source          1..11323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..19)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      31..220
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(259..279)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(259..277)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(298..314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(298..314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(311..333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    322..338
                     /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)."
     promoter        complement(join(346..352,353..370,371..376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    391..412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(529..546)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(700..1288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(780..799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1459..2250,2251..2319))
                     /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     2082..2101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2320..2424)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2492..2510
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2548..2570)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2670..2689
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2836..3023
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3024..3686
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(3946..4401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4083..4104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4295..4314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4527..4549
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4541..4558
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4542..4558
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4568..4587
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4568..4586
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5780..5800)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        6022..6463
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6022..6139
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6434..6457
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             6496..10599
                     /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             6496..10599
                     /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             6496..10599
                     /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             10612..10632
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10642..10698
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             10708..10728
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
       61 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
      121 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
      181 actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt caccgcggtg
      241 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
      301 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      361 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      421 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      481 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      721 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1141 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1741 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     2461 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2521 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2581 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2641 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2701 ccataaattc ccgttttaag agcttggtga gcgctaggag tcactgccag gtatcgtttg
     2761 aacacggcat tagtcaggga agtcataaca cagtcctttc ccgcaatttt ctttttctat
     2821 tactcttggc ctcctctagt acactctata tttttttatg cctcggtaat gattttcatt
     2881 tttttttttc cacctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
     2941 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
     3001 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
     3061 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
     3121 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
     3181 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
     3241 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
     3301 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctaggg gccgtgcgtg
     3361 gagtaaaaag gtttggatca ggatttgcgc ctttggatga ggcactttcc agagcggtgg
     3421 tagatctttc gaacaggccg tacgcagttg tcgaacttgg tttgcaaagg gagaaagtag
     3481 gagatctctc ttgcgagatg atcccgcatt ttcttgaaag ctttgcagag gctagcagaa
     3541 ttaccctcca cgttgattgt ctgcgaggca agaatgatca tcaccgtagt gagagtgcgt
     3601 tcaaggctct tgcggttgcc ataagagaag ccacctcgcc caatggtacc aacgatgttc
     3661 cctccaccaa aggtgttctt atgtagtgac accgattatt taaagctgca gcatacgata
     3721 tatatacatg tgtatatatg tatacctatg aatgtcagta agtatgtata cgaacagtat
     3781 gatactgaag atgacaaggt aatgcatcat tctatacgtg tcattctgaa cgaggcgcgc
     3841 tttccttttt tctttttgct ttttcttttt ttttctcttg aactcgacgg atctatgcgg
     3901 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta
     3961 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4021 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4081 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4141 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4201 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4261 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4321 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4381 atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     4441 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     4501 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     4561 gcgcgcgtaa tacgactcac tatagggcga attgcatatc tttccataga ttttcaattt
     4621 ttgatgtctc catggtggta cagagaactt gtaaacaatt cggtccctac atgtgaggaa
     4681 attcgctgtg acacttttat cactgaactc caaatttaaa aaatagcata aaattcgtta
     4741 tacagcaaat ctatgtgttg caattaagaa ctaaaagata tagagtgcat attttcaaga
     4801 aggatagtaa gctggcaaat caagtcttag ccttgttcaa tgggttctta ccgtacttct
     4861 tttccaactt agagatgtag gtaatcaatc ttctgtggga cttagaatca acggacaaga
     4921 tgaaagacaa ccaacctcta acatgttcga tttcggagga tttaccgcag aaaatatggt
     4981 ggatcttggc tctgatttct ttgtactttt ctctaccaat accaaccttt tcttgggaga
     5041 taaccaaacc ggtaactttt ctttgggatc ttggaccaga aatacaggtc tttttggagt
     5101 tgatgaccaa accttctgat ggaataatgg agaacaagaa gtctctggcc ttaacaactt
     5161 tcttcataga ttgagcagac aaggtcaagt catcagcgta tctagtgtat atcaaacctc
     5221 tagaaccagc ataaccttga attctgtagt ccaacttgga gcaaatcaag ttagccaatt
     5281 ttggagaaga tggagcacct tgtggcaaca agttcttgta gcagcaaatc ttggtcaaaa
     5341 cagaggagat caatctgttg taacccaaag aatggaaaac accgaaaacc ttgttagcag
     5401 tcaaagatgg gaagaaatct tccaagtcga tgttcaaaat gaagttggca ccaatatgtg
     5461 gagtggcatt gttcaagatg gattggtgct tttcaaaacc gatagaaaat ggggaggagg
     5521 ataacttgtc caagatgttt ctcaaaaccc aaccttgtaa agccttcaat tctctggatg
     5581 gttggtagat agttctcatt ctcttttctg gacccttctt ttcaacagtg tagattctgt
     5641 atctgaaatc ggcggtgtag atcaataatc tcaaggtttc aacagaaatt ctggtagcct
     5701 tagacatatc gtgcaagttg ttcatgactg gcaaacccaa atttctcaat ctgaaggtgt
     5761 tcaagtattc agcggacttg acctttcttt tcttctttgg tggcatttat attgaatttt
     5821 caaaaattct tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg
     5881 cattaccacc atatacatat ccatatctaa tcttacttat atgttgtgga aatgtaaaga
     5941 gccccattat cttagcctaa aaaaaccttc tctttggaac tttcagtaat acgcttaact
     6001 gctcattgct atattgaagt acggattaga agccgccgag cgggcgacag ccctccgacg
     6061 gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc
     6121 ctcgcgccgc actgctccga acaataaaga ttctacaata ctagctttta tggttatgaa
     6181 gaggaaaaat tggcagtaac ctggccccac aaaccttcaa attaacgaat caaattaaca
     6241 accataggat gataatgcga ttagtttttt agccttattt ctggggtaat taatcagcga
     6301 agcgatgatt tttgatctat taacagatat ataaatggaa aagctgcata accactttaa
     6361 ctaatacttt caacattttc agtttgtatt acttcttatt caaatgtcat aaaagtatca
     6421 acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaaaaac aactagtgga
     6481 tcccccggga aaaaaatgga caagaagtac tccattgggc tcgatatcgg cacaaacagc
     6541 gtcggttggg ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt caaagttctg
     6601 ggcaataccg atcgccacag cataaagaag aacctcattg gcgccctcct gttcgactcc
     6661 ggggagacgg ccgaagccac gcggctcaaa agaacagcac ggcgcagata tacccgcaga
     6721 aagaatcgga tctgctacct gcaggagatc tttagtaatg agatggctaa ggtggatgac
     6781 tctttcttcc ataggctgga ggagtccttt ttggtggagg aggataaaaa gcacgagcgc
     6841 cacccaatct ttggcaatat cgtggacgag gtggcgtacc atgaaaagta cccaaccata
     6901 tatcatctga ggaagaagct tgtagacagt actgataagg ctgacttgcg gttgatctat
     6961 ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc tcatcgaggg ggacctgaac
     7021 ccagacaaca gcgatgtcga caaactcttt atccaactgg ttcagactta caatcagctt
     7081 ttcgaagaga acccgatcaa cgcatccgga gttgacgcca aagcaatcct gagcgctagg
     7141 ctgtccaaat cccggcggct cgaaaacctc atcgcacagc tccctgggga gaagaagaac
     7201 ggcctgtttg gtaatcttat cgccctgtca ctcgggctga cccccaactt taaatctaac
     7261 ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag acacctacga tgatgatctc
     7321 gacaatctgc tggcccagat cggcgaccag tacgcagacc tttttttggc ggcaaagaac
     7381 ctgtcagacg ccattctgct gagtgatatt ctgcgagtga acacggagat caccaaagct
     7441 ccgctgagcg ctagtatgat caagcgctat gatgagcacc accaagactt gactttgctg
     7501 aaggcccttg tcagacagca actgcctgag aagtacaagg aaattttctt cgatcagtct
     7561 aaaaatggct acgccggata cattgacggc ggagcaagcc aggaggaatt ttacaaattt
     7621 attaagccca tcttggaaaa aatggacggc accgaggagc tgctggtaaa gcttaacaga
     7681 gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa gcatccccca ccagattcac
     7741 ctgggcgaac tgcacgctat cctcaggcgg caagaggatt tctacccctt tttgaaagat
     7801 aacagggaaa agattgagaa aatcctcaca tttcggatac cctactatgt aggccccctc
     7861 gcccggggaa attccagatt cgcgtggatg actcgcaaat cagaagagac catcactccc
     7921 tggaacttcg aggaagtcgt ggataagggg gcctctgccc agtccttcat cgaaaggatg
     7981 actaactttg ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac
     8041 gagtacttca cagtttataa cgagctcacc aaggtcaaat acgtcacaga agggatgaga
     8101 aagccagcat tcctgtctgg agagcagaag aaagctatcg tggacctcct cttcaagacg
     8161 aaccggaaag ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat tgaatgtttc
     8221 gactctgttg aaatcagcgg agtggaggat cgcttcaacg catccctggg aacgtatcac
     8281 gatctcctga aaatcattaa agacaaggac ttcctggaca atgaggagaa cgaggacatt
     8341 cttgaggaca ttgtcctcac ccttacgttg tttgaagata gggagatgat tgaagaacgc
     8401 ttgaaaactt acgctcatct cttcgacgac aaagtcatga aacagctcaa gaggcgccga
     8461 tatacaggat gggggcggct gtcaagaaaa ctgatcaatg ggatccgaga caagcagagt
     8521 ggaaagacaa tcctggattt tcttaagtcc gatggatttg ccaaccggaa cttcatgcag
     8581 ttgatccatg atgactctct cacctttaag gaggacatcc agaaagcaca agtttctggc
     8641 cagggggaca gtcttcacga gcacatcgct aatcttgcag gtagcccagc tatcaaaaag
     8701 ggaatactgc agaccgttaa ggtcgtggat gaactcgtca aagtaatggg aaggcataag
     8761 cccgagaata tcgttatcga gatggcccga gagaaccaaa ctacccagaa gggacagaag
     8821 aacagtaggg aaaggatgaa gaggattgaa gagggtataa aagaactggg gtcccaaatc
     8881 cttaaggaac acccagttga aaacacccag cttcagaatg agaagctcta cctgtactac
     8941 ctgcagaacg gcagggacat gtacgtggat caggaactgg acatcaatcg gctctccgac
     9001 tacgacgtgg atcatatcgt gccccagtct tttctcaaag atgattctat tgataataaa
     9061 gtgttgacaa gatccgataa aaatagaggg aagagtgata acgtcccctc agaagaagtt
     9121 gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat cacacaacgg
     9181 aagttcgata atctgactaa ggctgaacga ggtggcctgt ctgagttgga taaagccggc
     9241 ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca agcacgtggc ccaaattctc
     9301 gattcacgca tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt
     9361 attactctga agtctaagct ggtctcagat ttcagaaagg actttcagtt ttataaggtg
     9421 agagagatca acaattacca ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact
     9481 gcacttatca aaaaatatcc caagcttgaa tctgaatttg tttacggaga ctataaagtg
     9541 tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc caccgctaag
     9601 tacttctttt acagcaatat tatgaatttt ttcaagaccg agattacact ggccaatgga
     9661 gagattcgga agcgaccact tatcgaaaca aacggagaaa caggagaaat cgtgtgggac
     9721 aagggtaggg atttcgcgac agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt
     9781 aaaaagaccg aagtacagac cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac
     9841 agcgacaagc tgatcgcacg caaaaaagat tgggacccca agaaatacgg cggattcgat
     9901 tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa
     9961 aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc aagcttcgaa
    10021 aaaaacccca tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc
    10081 attaagcttc ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct
    10141 agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc
    10201 ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa tgagcagaag
    10261 cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgaa
    10321 ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat
    10381 aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg
    10441 accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagaaagcgg
    10501 tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc
    10561 tatgaaacaa gaatcgacct ctctcagctc ggtggagaca gcagggctga ccccaagaag
    10621 aagaggaagg tgggatccgg cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc
    10681 gaagagaatc ctggaccggc tagcatgcca aagaaaaaga gaaaggtctc ctctgatcac
    10741 tatttggata ttagattgag acctgatcca gagtttcctc cagctcaatt aatgtccgtt
    10801 ttgttcggta aattgcacca agctttggtc gctcaaggcg gtgacagaat cggtgtttct
    10861 ttcccagatt tggacgaatc tagatccaga ttgggtgaaa gattgagaat tcatgcttct
    10921 gctgatgact taagagcttt gttagccaga ccttggttgg aaggtttgag agaccacttg
    10981 caatttggtg agccagctgt tgtcccacat ccaacccctt atagacaagt ctccagagtt
    11041 caagctaagt ctaacccaga aagattgcgt agacgtttga tgcgtagaca cgacttgtct
    11101 gaagaggaag ctagaaaaag aattccagac accgtcgcca gagccttgga cttgcctttc
    11161 gtcaccttga gatctcaatc tactggtcaa catttcagat tgtttatcag acacggtcca
    11221 ctacaagtta ctgctgagga aggcggtttc acttgttacg gtttatctaa gggcggtttt
    11281 gtcccttggt tttgatctct tctcgagtca tgtaattagt tat
//
