LOCUS       sequence-376661-       10404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -Eco1 RT-P2A-Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10404)
  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 10404)
  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/s41589-021-00927-y"; journalName: "Nat Chem Biol"; date: 
            "2022-02"; volume: "18"; issue: "2"; pages: "199-206"
COMMENT     Sequence Label: pSCL.102
FEATURES             Location/Qualifiers
     source          1..10404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(12..32)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        254..695
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    254..371
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     666..689
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             734..754
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1712..1768
                     /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             1769..5869
                     /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             1769..5869
                     /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             1769..5869
                     /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             5882..5902
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5934..5952)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      5964..6153
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6192..6212)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6192..6210)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6231..6247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6231..6247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6244..6266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6255..6271
                     /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(6279..6285,6286..6303,6304..6309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6324..6345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6462..6479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6633..7221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6713..6732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7392..8183,8184..8252))
                     /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     8015..8034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8253..8357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8425..8443
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8481..8503)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8603..8622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        8769..8956
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             8957..9619
                     /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(9879..10334)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10016..10037)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10228..10247
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcagcggact tgacctttct tttcttcttt ggtggcattt atattgaatt ttcaaaaatt
       61 cttacttttt ttttggatgg acgcaaagaa gtttaataat catattacat ggcattacca
      121 ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa gagccccatt
      181 atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa ctgctcattg
      241 ctatattgaa gtacggatta gaagccgccg agcgggcgac agccctccga cggaagactc
      301 tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc
      361 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
      421 attggcagta acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg
      481 atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
      541 tttttgatct attaacagat atataaatgg aaaagctgca taaccacttt aactaatact
      601 ttcaacattt tcagtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa
      661 ttgttaatat acctctatac tttaacgtca aggagaaaaa acaactagtg gatcccccgg
      721 gaaaaaaatg ccaccaaaga agaaaagaaa ggtcaagtcc gctgaatact tgaacacctt
      781 cagattgaga aatttgggtt tgccagtcat gaacaacttg cacgatatgt ctaaggctac
      841 cagaatttct gttgaaacct tgagattatt gatctacacc gccgatttca gatacagaat
      901 ctacactgtt gaaaagaagg gtccagaaaa gagaatgaga actatctacc aaccatccag
      961 agaattgaag gctttacaag gttgggtttt gagaaacatc ttggacaagt tatcctcctc
     1021 cccattttct atcggttttg aaaagcacca atccatcttg aacaatgcca ctccacatat
     1081 tggtgccaac ttcattttga acatcgactt ggaagatttc ttcccatctt tgactgctaa
     1141 caaggttttc ggtgttttcc attctttggg ttacaacaga ttgatctcct ctgttttgac
     1201 caagatttgc tgctacaaga acttgttgcc acaaggtgct ccatcttctc caaaattggc
     1261 taacttgatt tgctccaagt tggactacag aattcaaggt tatgctggtt ctagaggttt
     1321 gatatacact agatacgctg atgacttgac cttgtctgct caatctatga agaaagttgt
     1381 taaggccaga gacttcttgt tctccattat tccatcagaa ggtttggtca tcaactccaa
     1441 aaagacctgt atttctggtc caagatccca aagaaaagtt accggtttgg ttatctccca
     1501 agaaaaggtt ggtattggta gagaaaagta caaagaaatc agagccaaga tccaccatat
     1561 tttctgcggt aaatcctccg aaatcgaaca tgttagaggt tggttgtctt tcatcttgtc
     1621 cgttgattct aagtcccaca gaagattgat tacctacatc tctaagttgg aaaagaagta
     1681 cggtaagaac ccattgaaca aggctaagac tgcaacaaac ttctctctgc tgaaacaagc
     1741 cggagatgtc gaagagaatc ctggaccgga caagaagtac tccattgggc tcgatatcgg
     1801 cacaaacagc gtcggttggg ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt
     1861 caaagttctg ggcaataccg atcgccacag cataaagaag aacctcattg gcgccctcct
     1921 gttcgactcc ggggagacgg ccgaagccac gcggctcaaa agaacagcac ggcgcagata
     1981 tacccgcaga aagaatcgga tctgctacct gcaggagatc tttagtaatg agatggctaa
     2041 ggtggatgac tctttcttcc ataggctgga ggagtccttt ttggtggagg aggataaaaa
     2101 gcacgagcgc cacccaatct ttggcaatat cgtggacgag gtggcgtacc atgaaaagta
     2161 cccaaccata tatcatctga ggaagaagct tgtagacagt actgataagg ctgacttgcg
     2221 gttgatctat ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc tcatcgaggg
     2281 ggacctgaac ccagacaaca gcgatgtcga caaactcttt atccaactgg ttcagactta
     2341 caatcagctt ttcgaagaga acccgatcaa cgcatccgga gttgacgcca aagcaatcct
     2401 gagcgctagg ctgtccaaat cccggcggct cgaaaacctc atcgcacagc tccctgggga
     2461 gaagaagaac ggcctgtttg gtaatcttat cgccctgtca ctcgggctga cccccaactt
     2521 taaatctaac ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag acacctacga
     2581 tgatgatctc gacaatctgc tggcccagat cggcgaccag tacgcagacc tttttttggc
     2641 ggcaaagaac ctgtcagacg ccattctgct gagtgatatt ctgcgagtga acacggagat
     2701 caccaaagct ccgctgagcg ctagtatgat caagcgctat gatgagcacc accaagactt
     2761 gactttgctg aaggcccttg tcagacagca actgcctgag aagtacaagg aaattttctt
     2821 cgatcagtct aaaaatggct acgccggata cattgacggc ggagcaagcc aggaggaatt
     2881 ttacaaattt attaagccca tcttggaaaa aatggacggc accgaggagc tgctggtaaa
     2941 gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa gcatccccca
     3001 ccagattcac ctgggcgaac tgcacgctat cctcaggcgg caagaggatt tctacccctt
     3061 tttgaaagat aacagggaaa agattgagaa aatcctcaca tttcggatac cctactatgt
     3121 aggccccctc gcccggggaa attccagatt cgcgtggatg actcgcaaat cagaagagac
     3181 catcactccc tggaacttcg aggaagtcgt ggataagggg gcctctgccc agtccttcat
     3241 cgaaaggatg actaactttg ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc
     3301 tctgctgtac gagtacttca cagtttataa cgagctcacc aaggtcaaat acgtcacaga
     3361 agggatgaga aagccagcat tcctgtctgg agagcagaag aaagctatcg tggacctcct
     3421 cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat
     3481 tgaatgtttc gactctgttg aaatcagcgg agtggaggat cgcttcaacg catccctggg
     3541 aacgtatcac gatctcctga aaatcattaa agacaaggac ttcctggaca atgaggagaa
     3601 cgaggacatt cttgaggaca ttgtcctcac ccttacgttg tttgaagata gggagatgat
     3661 tgaagaacgc ttgaaaactt acgctcatct cttcgacgac aaagtcatga aacagctcaa
     3721 gaggcgccga tatacaggat gggggcggct gtcaagaaaa ctgatcaatg ggatccgaga
     3781 caagcagagt ggaaagacaa tcctggattt tcttaagtcc gatggatttg ccaaccggaa
     3841 cttcatgcag ttgatccatg atgactctct cacctttaag gaggacatcc agaaagcaca
     3901 agtttctggc cagggggaca gtcttcacga gcacatcgct aatcttgcag gtagcccagc
     3961 tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat gaactcgtca aagtaatggg
     4021 aaggcataag cccgagaata tcgttatcga gatggcccga gagaaccaaa ctacccagaa
     4081 gggacagaag aacagtaggg aaaggatgaa gaggattgaa gagggtataa aagaactggg
     4141 gtcccaaatc cttaaggaac acccagttga aaacacccag cttcagaatg agaagctcta
     4201 cctgtactac ctgcagaacg gcagggacat gtacgtggat caggaactgg acatcaatcg
     4261 gctctccgac tacgacgtgg atcatatcgt gccccagtct tttctcaaag atgattctat
     4321 tgataataaa gtgttgacaa gatccgataa aaatagaggg aagagtgata acgtcccctc
     4381 agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat
     4441 cacacaacgg aagttcgata atctgactaa ggctgaacga ggtggcctgt ctgagttgga
     4501 taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca agcacgtggc
     4561 ccaaattctc gattcacgca tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga
     4621 ggtgaaagtt attactctga agtctaagct ggtctcagat ttcagaaagg actttcagtt
     4681 ttataaggtg agagagatca acaattacca ccatgcgcat gatgcctacc tgaatgcagt
     4741 ggtaggcact gcacttatca aaaaatatcc caagcttgaa tctgaatttg tttacggaga
     4801 ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc
     4861 caccgctaag tacttctttt acagcaatat tatgaatttt ttcaagaccg agattacact
     4921 ggccaatgga gagattcgga agcgaccact tatcgaaaca aacggagaaa caggagaaat
     4981 cgtgtgggac aagggtaggg atttcgcgac agtccggaag gtcctgtcca tgccgcaggt
     5041 gaacatcgtt aaaaagaccg aagtacagac cggaggcttc tccaaggaaa gtatcctccc
     5101 gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat tgggacccca agaaatacgg
     5161 cggattcgat tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg
     5221 gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc
     5281 aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa
     5341 agacctcatc attaagcttc ccaagtactc tctctttgag cttgaaaacg gccggaaacg
     5401 aatgctcgct agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata
     5461 cgttaatttc ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa
     5521 tgagcagaag cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca
     5581 aataagcgaa ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc
     5641 tgcttacaat aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatccactt
     5701 gtttactctg accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga
     5761 cagaaagcgg tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat
     5821 tacggggctc tatgaaacaa gaatcgacct ctctcagctc ggtggagaca gcagggctga
     5881 ccccaagaag aagaggaagg tgtgatctct tctcgagtca tgtaattagt tatgtcacgc
     5941 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     6001 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     6061 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     6121 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttcaccgcg gtggagctcc
     6181 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     6241 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6301 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6421 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6481 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6541 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6601 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6661 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6721 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6781 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6841 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6901 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6961 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7021 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7081 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7141 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7201 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7261 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7321 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7381 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7441 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7501 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7561 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7621 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7681 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7741 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7801 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7861 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7921 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7981 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8041 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8101 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8161 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8221 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8281 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8341 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     8401 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     8461 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     8521 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     8581 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa
     8641 ttcccgtttt aagagcttgg tgagcgctag gagtcactgc caggtatcgt ttgaacacgg
     8701 cattagtcag ggaagtcata acacagtcct ttcccgcaat tttctttttc tattactctt
     8761 ggcctcctct agtacactct atattttttt atgcctcggt aatgattttc attttttttt
     8821 ttccacctag cggatgactc tttttttttc ttagcgattg gcattatcac ataatgaatt
     8881 atacattata taaagtaatg tgatttcttc gaagaatata ctaaaaaatg agcaggcaag
     8941 ataaacgaag gcaaagatga cagagcagaa agccctagta aagcgtatta caaatgaaac
     9001 caagattcag attgcgatct ctttaaaggg tggtccccta gcgatagagc actcgatctt
     9061 cccagaaaaa gaggcagaag cagtagcaga acaggccaca caatcgcaag tgattaacgt
     9121 ccacacaggt atagggtttc tggaccatat gatacatgct ctggccaagc attccggctg
     9181 gtcgctaatc gttgagtgca ttggtgactt acacatagac gaccatcaca ccactgaaga
     9241 ctgcgggatt gctctcggtc aagcttttaa agaggcccta ggggccgtgc gtggagtaaa
     9301 aaggtttgga tcaggatttg cgcctttgga tgaggcactt tccagagcgg tggtagatct
     9361 ttcgaacagg ccgtacgcag ttgtcgaact tggtttgcaa agggagaaag taggagatct
     9421 ctcttgcgag atgatcccgc attttcttga aagctttgca gaggctagca gaattaccct
     9481 ccacgttgat tgtctgcgag gcaagaatga tcatcaccgt agtgagagtg cgttcaaggc
     9541 tcttgcggtt gccataagag aagccacctc gcccaatggt accaacgatg ttccctccac
     9601 caaaggtgtt cttatgtagt gacaccgatt atttaaagct gcagcatacg atatatatac
     9661 atgtgtatat atgtatacct atgaatgtca gtaagtatgt atacgaacag tatgatactg
     9721 aagatgacaa ggtaatgcat cattctatac gtgtcattct gaacgaggcg cgctttcctt
     9781 ttttcttttt gctttttctt tttttttctc ttgaactcga cggatctatg cggtgtgaaa
     9841 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     9901 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     9961 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
    10021 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
    10081 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
    10141 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
    10201 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
    10261 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
    10321 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
    10381 cgggcctctt cgctattacg ccag
//
