LOCUS       sequence-376616-       10446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -Eco1 RT/Cas9 fusion, using linker2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10446)
  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 10446)
  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.94
FEATURES             Location/Qualifiers
     source          1..10446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             32..4132
                     /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             32..4132
                     /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             32..4132
                     /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             4145..4165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4197..4215)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      4227..4416
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4455..4475)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4455..4473)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4494..4510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4494..4510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4507..4529)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4518..4534
                     /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(4542..4548,4549..4566,4567..4572))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4587..4608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4725..4742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4896..5484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4976..4995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5655..6446,6447..6515))
                     /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     6278..6297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6516..6620)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6688..6706
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6744..6766)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6866..6885
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7032..7219
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             7220..7882
                     /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(8142..8597)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8279..8300)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8491..8510
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(8679..8699)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        8921..9362
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    8921..9038
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     9333..9356
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             9401..9421
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gagcagtggc ggcagcagcg gcggcagcag cgacaagaag tactccattg ggctcgatat
       61 cggcacaaac agcgtcggtt gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa
      121 attcaaagtt ctgggcaata ccgatcgcca cagcataaag aagaacctca ttggcgccct
      181 cctgttcgac tccggggaga cggccgaagc cacgcggctc aaaagaacag cacggcgcag
      241 atatacccgc agaaagaatc ggatctgcta cctgcaggag atctttagta atgagatggc
      301 taaggtggat gactctttct tccataggct ggaggagtcc tttttggtgg aggaggataa
      361 aaagcacgag cgccacccaa tctttggcaa tatcgtggac gaggtggcgt accatgaaaa
      421 gtacccaacc atatatcatc tgaggaagaa gcttgtagac agtactgata aggctgactt
      481 gcggttgatc tatctcgcgc tggcgcatat gatcaaattt cggggacact tcctcatcga
      541 gggggacctg aacccagaca acagcgatgt cgacaaactc tttatccaac tggttcagac
      601 ttacaatcag cttttcgaag agaacccgat caacgcatcc ggagttgacg ccaaagcaat
      661 cctgagcgct aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac agctccctgg
      721 ggagaagaag aacggcctgt ttggtaatct tatcgccctg tcactcgggc tgacccccaa
      781 ctttaaatct aacttcgacc tggccgaaga tgccaagctt caactgagca aagacaccta
      841 cgatgatgat ctcgacaatc tgctggccca gatcggcgac cagtacgcag accttttttt
      901 ggcggcaaag aacctgtcag acgccattct gctgagtgat attctgcgag tgaacacgga
      961 gatcaccaaa gctccgctga gcgctagtat gatcaagcgc tatgatgagc accaccaaga
     1021 cttgactttg ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca aggaaatttt
     1081 cttcgatcag tctaaaaatg gctacgccgg atacattgac ggcggagcaa gccaggagga
     1141 attttacaaa tttattaagc ccatcttgga aaaaatggac ggcaccgagg agctgctggt
     1201 aaagcttaac agagaagatc tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc
     1261 ccaccagatt cacctgggcg aactgcacgc tatcctcagg cggcaagagg atttctaccc
     1321 ctttttgaaa gataacaggg aaaagattga gaaaatcctc acatttcgga taccctacta
     1381 tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg atgactcgca aatcagaaga
     1441 gaccatcact ccctggaact tcgaggaagt cgtggataag ggggcctctg cccagtcctt
     1501 catcgaaagg atgactaact ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca
     1561 ctctctgctg tacgagtact tcacagttta taacgagctc accaaggtca aatacgtcac
     1621 agaagggatg agaaagccag cattcctgtc tggagagcag aagaaagcta tcgtggacct
     1681 cctcttcaag acgaaccgga aagttaccgt gaaacagctc aaagaagact atttcaaaaa
     1741 gattgaatgt ttcgactctg ttgaaatcag cggagtggag gatcgcttca acgcatccct
     1801 gggaacgtat cacgatctcc tgaaaatcat taaagacaag gacttcctgg acaatgagga
     1861 gaacgaggac attcttgagg acattgtcct cacccttacg ttgtttgaag atagggagat
     1921 gattgaagaa cgcttgaaaa cttacgctca tctcttcgac gacaaagtca tgaaacagct
     1981 caagaggcgc cgatatacag gatgggggcg gctgtcaaga aaactgatca atgggatccg
     2041 agacaagcag agtggaaaga caatcctgga ttttcttaag tccgatggat ttgccaaccg
     2101 gaacttcatg cagttgatcc atgatgactc tctcaccttt aaggaggaca tccagaaagc
     2161 acaagtttct ggccaggggg acagtcttca cgagcacatc gctaatcttg caggtagccc
     2221 agctatcaaa aagggaatac tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat
     2281 gggaaggcat aagcccgaga atatcgttat cgagatggcc cgagagaacc aaactaccca
     2341 gaagggacag aagaacagta gggaaaggat gaagaggatt gaagagggta taaaagaact
     2401 ggggtcccaa atccttaagg aacacccagt tgaaaacacc cagcttcaga atgagaagct
     2461 ctacctgtac tacctgcaga acggcaggga catgtacgtg gatcaggaac tggacatcaa
     2521 tcggctctcc gactacgacg tggatcatat cgtgccccag tcttttctca aagatgattc
     2581 tattgataat aaagtgttga caagatccga taaaaataga gggaagagtg ataacgtccc
     2641 ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga acgccaaact
     2701 gatcacacaa cggaagttcg ataatctgac taaggctgaa cgaggtggcc tgtctgagtt
     2761 ggataaagcc ggcttcatca aaaggcagct tgttgagaca cgccagatca ccaagcacgt
     2821 ggcccaaatt ctcgattcac gcatgaacac caagtacgat gaaaatgaca aactgattcg
     2881 agaggtgaaa gttattactc tgaagtctaa gctggtctca gatttcagaa aggactttca
     2941 gttttataag gtgagagaga tcaacaatta ccaccatgcg catgatgcct acctgaatgc
     3001 agtggtaggc actgcactta tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg
     3061 agactataaa gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa
     3121 ggccaccgct aagtacttct tttacagcaa tattatgaat tttttcaaga ccgagattac
     3181 actggccaat ggagagattc ggaagcgacc acttatcgaa acaaacggag aaacaggaga
     3241 aatcgtgtgg gacaagggta gggatttcgc gacagtccgg aaggtcctgt ccatgccgca
     3301 ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg aaagtatcct
     3361 cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa gattgggacc ccaagaaata
     3421 cggcggattc gattctccta cagtcgctta cagtgtactg gttgtggcca aagtggagaa
     3481 agggaagtct aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg
     3541 atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata aagaggtcaa
     3601 aaaagacctc atcattaagc ttcccaagta ctctctcttt gagcttgaaa acggccggaa
     3661 acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac gagctggcac tgccctctaa
     3721 atacgttaat ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga
     3781 taatgagcag aagcagctgt tcgtggaaca acacaaacac taccttgatg agatcatcga
     3841 gcaaataagc gaattctcca aaagagtgat cctcgccgac gctaacctcg ataaggtgct
     3901 ttctgcttac aataagcaca gggataagcc catcagggag caggcagaaa acattatcca
     3961 cttgtttact ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg acaccaccat
     4021 agacagaaag cggtacacct ctacaaagga ggtcctggac gccacactga ttcatcagtc
     4081 aattacgggg ctctatgaaa caagaatcga cctctctcag ctcggtggag acagcagggc
     4141 tgaccccaag aagaagagga aggtgtgatc tcttctcgag tcatgtaatt agttatgtca
     4201 cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa
     4261 cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt
     4321 atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg
     4381 aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttcacc gcggtggagc
     4441 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4501 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4561 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4621 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4681 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4741 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4801 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4861 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4921 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4981 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5041 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5101 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5161 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5221 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5281 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5341 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5401 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5461 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5521 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5581 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5641 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5701 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5761 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5821 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5881 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5941 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6001 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6061 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6121 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6181 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6241 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6301 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6361 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6421 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6481 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6541 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6601 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6661 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6721 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     6781 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     6841 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     6901 aaattcccgt tttaagagct tggtgagcgc taggagtcac tgccaggtat cgtttgaaca
     6961 cggcattagt cagggaagtc ataacacagt cctttcccgc aattttcttt ttctattact
     7021 cttggcctcc tctagtacac tctatatttt tttatgcctc ggtaatgatt ttcatttttt
     7081 tttttccacc tagcggatga ctcttttttt ttcttagcga ttggcattat cacataatga
     7141 attatacatt atataaagta atgtgatttc ttcgaagaat atactaaaaa atgagcaggc
     7201 aagataaacg aaggcaaaga tgacagagca gaaagcccta gtaaagcgta ttacaaatga
     7261 aaccaagatt cagattgcga tctctttaaa gggtggtccc ctagcgatag agcactcgat
     7321 cttcccagaa aaagaggcag aagcagtagc agaacaggcc acacaatcgc aagtgattaa
     7381 cgtccacaca ggtatagggt ttctggacca tatgatacat gctctggcca agcattccgg
     7441 ctggtcgcta atcgttgagt gcattggtga cttacacata gacgaccatc acaccactga
     7501 agactgcggg attgctctcg gtcaagcttt taaagaggcc ctaggggccg tgcgtggagt
     7561 aaaaaggttt ggatcaggat ttgcgccttt ggatgaggca ctttccagag cggtggtaga
     7621 tctttcgaac aggccgtacg cagttgtcga acttggtttg caaagggaga aagtaggaga
     7681 tctctcttgc gagatgatcc cgcattttct tgaaagcttt gcagaggcta gcagaattac
     7741 cctccacgtt gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga gtgcgttcaa
     7801 ggctcttgcg gttgccataa gagaagccac ctcgcccaat ggtaccaacg atgttccctc
     7861 caccaaaggt gttcttatgt agtgacaccg attatttaaa gctgcagcat acgatatata
     7921 tacatgtgta tatatgtata cctatgaatg tcagtaagta tgtatacgaa cagtatgata
     7981 ctgaagatga caaggtaatg catcattcta tacgtgtcat tctgaacgag gcgcgctttc
     8041 cttttttctt tttgcttttt cttttttttt ctcttgaact cgacggatct atgcggtgtg
     8101 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     8161 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     8221 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     8281 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     8341 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     8401 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     8461 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     8521 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     8581 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     8641 gtgcgggcct cttcgctatt acgccagtca gcggacttga cctttctttt cttctttggt
     8701 ggcatttata ttgaattttc aaaaattctt actttttttt tggatggacg caaagaagtt
     8761 taataatcat attacatggc attaccacca tatacatatc catatctaat cttacttata
     8821 tgttgtggaa atgtaaagag ccccattatc ttagcctaaa aaaaccttct ctttggaact
     8881 ttcagtaata cgcttaactg ctcattgcta tattgaagta cggattagaa gccgccgagc
     8941 gggcgacagc cctccgacgg aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt
     9001 tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa caataaagat tctacaatac
     9061 tagcttttat ggttatgaag aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa
     9121 ttaacgaatc aaattaacaa ccataggatg ataatgcgat tagtttttta gccttatttc
     9181 tggggtaatt aatcagcgaa gcgatgattt ttgatctatt aacagatata taaatggaaa
     9241 agctgcataa ccactttaac taatactttc aacattttca gtttgtatta cttcttattc
     9301 aaatgtcata aaagtatcaa caaaaaattg ttaatatacc tctatacttt aacgtcaagg
     9361 agaaaaaaca actagtggat cccccgggaa aaaaatgcca ccaaagaaga aaagaaaggt
     9421 caagtccgct gaatacttga acaccttcag attgagaaat ttgggtttgc cagtcatgaa
     9481 caacttgcac gatatgtcta aggctaccag aatttctgtt gaaaccttga gattattgat
     9541 ctacaccgcc gatttcagat acagaatcta cactgttgaa aagaagggtc cagaaaagag
     9601 aatgagaact atctaccaac catccagaga attgaaggct ttacaaggtt gggttttgag
     9661 aaacatcttg gacaagttat cctcctcccc attttctatc ggttttgaaa agcaccaatc
     9721 catcttgaac aatgccactc cacatattgg tgccaacttc attttgaaca tcgacttgga
     9781 agatttcttc ccatctttga ctgctaacaa ggttttcggt gttttccatt ctttgggtta
     9841 caacagattg atctcctctg ttttgaccaa gatttgctgc tacaagaact tgttgccaca
     9901 aggtgctcca tcttctccaa aattggctaa cttgatttgc tccaagttgg actacagaat
     9961 tcaaggttat gctggttcta gaggtttgat atacactaga tacgctgatg acttgacctt
    10021 gtctgctcaa tctatgaaga aagttgttaa ggccagagac ttcttgttct ccattattcc
    10081 atcagaaggt ttggtcatca actccaaaaa gacctgtatt tctggtccaa gatcccaaag
    10141 aaaagttacc ggtttggtta tctcccaaga aaaggttggt attggtagag aaaagtacaa
    10201 agaaatcaga gccaagatcc accatatttt ctgcggtaaa tcctccgaaa tcgaacatgt
    10261 tagaggttgg ttgtctttca tcttgtccgt tgattctaag tcccacagaa gattgattac
    10321 ctacatctct aagttggaaa agaagtacgg taagaaccca ttgaacaagg ctaagacttc
    10381 tggaggatct agcggaggat cctctggcag cgagacacca ggaacaagcg agtcagcaac
    10441 accaga
//
