LOCUS       sequence-376540-       10407 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express -Eco1 RT/Cas9 fusion, using linker1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10407)
  AUTHORS   Lopez SC, Crawford KD, Lear SK, Bhattarai-Kline S, Shipman SL
  TITLE     Precise genome editing across kingdoms of life using retron-derived 
            DNA.
  JOURNAL   Nat Chem Biol. 2022 Feb;18(2):199-206. doi: 
            10.1038/s41589-021-00927-y. Epub 2021 Dec 23.
  PUBMED    34949838
REFERENCE   2  (bases 1 to 10407)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41589-021-00927-y"; journalName: "Nat Chem Biol"; date: 
            "2022-02"; volume: "18"; issue: "2"; pages: "199-206"
COMMENT     Sequence Label: pSCL.71
FEATURES             Location/Qualifiers
     source          1..10407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             21..4121
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             21..4121
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             21..4121
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4134..4154
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4186..4204)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      4216..4405
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4444..4464)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4444..4462)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4483..4499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4483..4499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4496..4518)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4507..4523
                     /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(4531..4537,4538..4555,4556..4561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4576..4597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4714..4731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4885..5473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4965..4984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5644..6435,6436..6504))
                     /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     6267..6286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6505..6609)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6677..6695
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6733..6755)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6855..6874
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7021..7208
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             7209..7871
                     /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(8131..8586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8268..8289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8480..8499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(8668..8688)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        8910..9351
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    8910..9027
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     9322..9345
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             9390..9410
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctggcgcaac gagcggtggt gacaagaagt actccattgg gctcgatatc ggcacaaaca
       61 gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
      121 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
      181 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
      241 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
      301 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
      361 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
      421 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
      481 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
      541 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
      601 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
      661 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
      721 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
      781 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
      841 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
      901 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
      961 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     1021 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     1081 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     1141 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
     1201 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     1261 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     1321 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     1381 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     1441 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     1501 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     1561 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     1621 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     1681 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     1741 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     1801 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     1861 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     1921 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
     1981 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     2041 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     2101 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     2161 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     2221 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     2281 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     2341 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     2401 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     2461 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     2521 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
     2581 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     2641 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     2701 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     2761 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     2821 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     2881 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     2941 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     3001 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     3061 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     3121 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     3181 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     3241 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     3301 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     3361 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     3421 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     3481 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     3541 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     3601 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     3661 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     3721 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     3781 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     3841 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     3901 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     3961 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     4021 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     4081 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
     4141 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacattc
     4201 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     4261 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     4321 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     4381 tgagaaggtt ttgggacgct cgaaggcttt aatttcaccg cggtggagct ccagcttttg
     4441 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     4501 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4561 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4621 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4681 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4741 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4801 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4861 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4921 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4981 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5041 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5101 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5161 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5221 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5281 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5341 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5401 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5461 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5521 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5581 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5641 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5701 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5761 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5821 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5881 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5941 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6001 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6061 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6121 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6181 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6241 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6301 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6361 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6421 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6481 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6541 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6601 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6661 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6721 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6781 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6841 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aattcccgtt
     6901 ttaagagctt ggtgagcgct aggagtcact gccaggtatc gtttgaacac ggcattagtc
     6961 agggaagtca taacacagtc ctttcccgca attttctttt tctattactc ttggcctcct
     7021 ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt ttttccacct
     7081 agcggatgac tctttttttt tcttagcgat tggcattatc acataatgaa ttatacatta
     7141 tataaagtaa tgtgatttct tcgaagaata tactaaaaaa tgagcaggca agataaacga
     7201 aggcaaagat gacagagcag aaagccctag taaagcgtat tacaaatgaa accaagattc
     7261 agattgcgat ctctttaaag ggtggtcccc tagcgataga gcactcgatc ttcccagaaa
     7321 aagaggcaga agcagtagca gaacaggcca cacaatcgca agtgattaac gtccacacag
     7381 gtatagggtt tctggaccat atgatacatg ctctggccaa gcattccggc tggtcgctaa
     7441 tcgttgagtg cattggtgac ttacacatag acgaccatca caccactgaa gactgcggga
     7501 ttgctctcgg tcaagctttt aaagaggccc taggggccgt gcgtggagta aaaaggtttg
     7561 gatcaggatt tgcgcctttg gatgaggcac tttccagagc ggtggtagat ctttcgaaca
     7621 ggccgtacgc agttgtcgaa cttggtttgc aaagggagaa agtaggagat ctctcttgcg
     7681 agatgatccc gcattttctt gaaagctttg cagaggctag cagaattacc ctccacgttg
     7741 attgtctgcg aggcaagaat gatcatcacc gtagtgagag tgcgttcaag gctcttgcgg
     7801 ttgccataag agaagccacc tcgcccaatg gtaccaacga tgttccctcc accaaaggtg
     7861 ttcttatgta gtgacaccga ttatttaaag ctgcagcata cgatatatat acatgtgtat
     7921 atatgtatac ctatgaatgt cagtaagtat gtatacgaac agtatgatac tgaagatgac
     7981 aaggtaatgc atcattctat acgtgtcatt ctgaacgagg cgcgctttcc ttttttcttt
     8041 ttgctttttc tttttttttc tcttgaactc gacggatcta tgcggtgtga aataccgcac
     8101 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
     8161 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     8221 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     8281 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     8341 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     8401 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     8461 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     8521 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     8581 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     8641 ttcgctatta cgccagtcag cggacttgac ctttcttttc ttctttggtg gcatttatat
     8701 tgaattttca aaaattctta cttttttttt ggatggacgc aaagaagttt aataatcata
     8761 ttacatggca ttaccaccat atacatatcc atatctaatc ttacttatat gttgtggaaa
     8821 tgtaaagagc cccattatct tagcctaaaa aaaccttctc tttggaactt tcagtaatac
     8881 gcttaactgc tcattgctat attgaagtac ggattagaag ccgccgagcg ggcgacagcc
     8941 ctccgacgga agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc
     9001 agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact agcttttatg
     9061 gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat taacgaatca
     9121 aattaacaac cataggatga taatgcgatt agttttttag ccttatttct ggggtaatta
     9181 atcagcgaag cgatgatttt tgatctatta acagatatat aaatggaaaa gctgcataac
     9241 cactttaact aatactttca acattttcag tttgtattac ttcttattca aatgtcataa
     9301 aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga gaaaaaacaa
     9361 ctagtggatc ccccgggaaa aaaatgccac caaagaagaa aagaaaggtc aagtccgctg
     9421 aatacttgaa caccttcaga ttgagaaatt tgggtttgcc agtcatgaac aacttgcacg
     9481 atatgtctaa ggctaccaga atttctgttg aaaccttgag attattgatc tacaccgccg
     9541 atttcagata cagaatctac actgttgaaa agaagggtcc agaaaagaga atgagaacta
     9601 tctaccaacc atccagagaa ttgaaggctt tacaaggttg ggttttgaga aacatcttgg
     9661 acaagttatc ctcctcccca ttttctatcg gttttgaaaa gcaccaatcc atcttgaaca
     9721 atgccactcc acatattggt gccaacttca ttttgaacat cgacttggaa gatttcttcc
     9781 catctttgac tgctaacaag gttttcggtg ttttccattc tttgggttac aacagattga
     9841 tctcctctgt tttgaccaag atttgctgct acaagaactt gttgccacaa ggtgctccat
     9901 cttctccaaa attggctaac ttgatttgct ccaagttgga ctacagaatt caaggttatg
     9961 ctggttctag aggtttgata tacactagat acgctgatga cttgaccttg tctgctcaat
    10021 ctatgaagaa agttgttaag gccagagact tcttgttctc cattattcca tcagaaggtt
    10081 tggtcatcaa ctccaaaaag acctgtattt ctggtccaag atcccaaaga aaagttaccg
    10141 gtttggttat ctcccaagaa aaggttggta ttggtagaga aaagtacaaa gaaatcagag
    10201 ccaagatcca ccatattttc tgcggtaaat cctccgaaat cgaacatgtt agaggttggt
    10261 tgtctttcat cttgtccgtt gattctaagt cccacagaag attgattacc tacatctcta
    10321 agttggaaaa gaagtacggt aagaacccat tgaacaaggc taagactggc ggtacgtctt
    10381 ccggtggctc tggtcccgct ggtagct
//
