LOCUS       sequence-439430-       10936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses site 2/Cas9/Eco1 RT in HIS3 locus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10936)
  AUTHORS   Crawford KD, Khan AG, Lopez SC, Goodarzi H, Shipman SL
  TITLE     High throughput variant libraries and machine learning yield design 
            rules for retron gene editors.
  JOURNAL   Nucleic Acids Res. 2025 Jan 11;53(2):gkae1199. doi: 
            10.1093/nar/gkae1199.
  PUBMED    39658047
REFERENCE   2  (bases 1 to 10936)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2025-01-11"; volume: "53(2):gkae1199. doi"; 
            pages: " 10.1093/nar/gkae1199"
COMMENT     Sequence Label: pSCL195
FEATURES             Location/Qualifiers
     source          1..10936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(301..756)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(438..459)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     650..669
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     882..904
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     896..913
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     897..913
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     923..942
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        923..941
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2135..2155)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        2377..2818
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2377..2494
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2789..2812
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2851..6954
                     /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             2851..6954
                     /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             2851..6954
                     /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             6967..6987
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7019..7037)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      7049..7238
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7277..7297)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7277..7295)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7316..7332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7316..7332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7329..7351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7340..7356
                     /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(7364..7370,7371..7388,7389..7394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7409..7430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7547..7564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7718..8306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7798..7817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8477..9268,9269..9337))
                     /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     9100..9119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9338..9442)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9783..9801
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     9961..9980
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        10127..10314
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             join(10315..10936,1..41)
                     /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"
ORIGIN
        1 gtaccaacga tgttccctcc accaaaggtg ttcttatgta gtgacaccga ttatttaaag
       61 ctgcagcata cgatatatat acatgtgtat atatgtatac ctatgaatgt cagtaagtat
      121 gtatacgaac agtatgatac tgaagatgac aaggtaatgc atcattctat acgtgtcatt
      181 ctgaacgagg cgcgctttcc ttttttcttt ttgctttttc tttttttttc tcttgaactc
      241 gacggatcta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
      301 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
      361 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      421 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      481 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      541 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      601 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      661 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      721 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa
      781 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      841 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      901 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgc atatctttcc
      961 atagattttc aatttttgat gtctccatgg tggtacagag aacttgtaaa caattcggtc
     1021 cctacatgtg aggaaattcg ctgtgacact tttatcactg aactccaaat ttaaaaaata
     1081 gcataaaatt cgttatacag caaatctatg tgttgcaatt aagaactaaa agatatagag
     1141 tgcatatttt caagaaggat agtaagctgg caaatcaagt cttagccttg ttcaatgggt
     1201 tcttaccgta cttcttttcc aacttagaga tgtaggtaat caatcttctg tgggacttag
     1261 aatcaacgga caagatgaaa gacaaccaac ctctaacatg ttcgatttcg gaggatttac
     1321 cgcagaaaat atggtggatc ttggctctga tttctttgta cttttctcta ccaataccaa
     1381 ccttttcttg ggagataacc aaaccggtaa cttttctttg ggatcttgga ccagaaatac
     1441 aggtcttttt ggagttgatg accaaacctt ctgatggaat aatggagaac aagaagtctc
     1501 tggccttaac aactttcttc atagattgag cagacaaggt caagtcatca gcgtatctag
     1561 tgtatatcaa acctctagaa ccagcataac cttgaattct gtagtccaac ttggagcaaa
     1621 tcaagttagc caattttgga gaagatggag caccttgtgg caacaagttc ttgtagcagc
     1681 aaatcttggt caaaacagag gagatcaatc tgttgtaacc caaagaatgg aaaacaccga
     1741 aaaccttgtt agcagtcaaa gatgggaaga aatcttccaa gtcgatgttc aaaatgaagt
     1801 tggcaccaat atgtggagtg gcattgttca agatggattg gtgcttttca aaaccgatag
     1861 aaaatgggga ggaggataac ttgtccaaga tgtttctcaa aacccaacct tgtaaagcct
     1921 tcaattctct ggatggttgg tagatagttc tcattctctt ttctggaccc ttcttttcaa
     1981 cagtgtagat tctgtatctg aaatcggcgg tgtagatcaa taatctcaag gtttcaacag
     2041 aaattctggt agccttagac atatcgtgca agttgttcat gactggcaaa cccaaatttc
     2101 tcaatctgaa ggtgttcaag tattcagcgg acttgacctt tcttttcttc tttggtggca
     2161 tttatattga attttcaaaa attcttactt tttttttgga tggacgcaaa gaagtttaat
     2221 aatcatatta catggcatta ccaccatata catatccata tctaatctta cttatatgtt
     2281 gtggaaatgt aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca
     2341 gtaatacgct taactgctca ttgctatatt gaagtacgga ttagaagccg ccgagcgggc
     2401 gacagccctc cgacggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct
     2461 gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc
     2521 ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaattaa
     2581 cgaatcaaat taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg
     2641 gtaattaatc agcgaagcga tgatttttga tctattaaca gatatataaa tggaaaagct
     2701 gcataaccac tttaactaat actttcaaca ttttcagttt gtattacttc ttattcaaat
     2761 gtcataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa
     2821 aaaacaacta gtggatcccc cgggaaaaaa atggacaaga agtactccat tgggctcgat
     2881 atcggcacaa acagcgtcgg ttgggccgtc attacggacg agtacaaggt gccgagcaaa
     2941 aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct cattggcgcc
     3001 ctcctgttcg actccgggga gacggccgaa gccacgcggc tcaaaagaac agcacggcgc
     3061 agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag taatgagatg
     3121 gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt ggaggaggat
     3181 aaaaagcacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc gtaccatgaa
     3241 aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga taaggctgac
     3301 ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca cttcctcatc
     3361 gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca actggttcag
     3421 acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga cgccaaagca
     3481 atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc acagctccct
     3541 ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg gctgaccccc
     3601 aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag caaagacacc
     3661 tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc agaccttttt
     3721 ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg agtgaacacg
     3781 gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga gcaccaccaa
     3841 gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta caaggaaatt
     3901 ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc aagccaggag
     3961 gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga ggagctgctg
     4021 gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc
     4081 ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga ggatttctac
     4141 ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg gataccctac
     4201 tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg caaatcagaa
     4261 gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc tgcccagtcc
     4321 ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt gcttcctaaa
     4381 cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt caaatacgtc
     4441 acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc tatcgtggac
     4501 ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa
     4561 aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt caacgcatcc
     4621 ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct ggacaatgag
     4681 gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga agatagggag
     4741 atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt catgaaacag
     4801 ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat caatgggatc
     4861 cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg atttgccaac
     4921 cggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga catccagaaa
     4981 gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct tgcaggtagc
     5041 ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact cgtcaaagta
     5101 atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa ccaaactacc
     5161 cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg tataaaagaa
     5221 ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca gaatgagaag
     5281 ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga actggacatc
     5341 aatcggctct ccgactacga cgtggatcat atcgtgcccc agtcttttct caaagatgat
     5401 tctattgata ataaagtgtt gacaagatcc gataaaaata gagggaagag tgataacgtc
     5461 ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct gaacgccaaa
     5521 ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg cctgtctgag
     5581 ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat caccaagcac
     5641 gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga caaactgatt
     5701 cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag aaaggacttt
     5761 cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc ctacctgaat
     5821 gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga atttgtttac
     5881 ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca ggaaataggc
     5941 aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa gaccgagatt
     6001 acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg agaaacagga
     6061 gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct gtccatgccg
     6121 caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa ggaaagtatc
     6181 ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga ccccaagaaa
     6241 tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc caaagtggag
     6301 aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac aatcatggag
     6361 cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata taaagaggtc
     6421 aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga aaacggccgg
     6481 aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc actgccctct
     6541 aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg gtctcccgaa
     6601 gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga tgagatcatc
     6661 gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct cgataaggtg
     6721 ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga aaacattatc
     6781 cacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt cgacaccacc
     6841 atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact gattcatcag
     6901 tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg agacagcagg
     6961 gctgacccca agaagaagag gaaggtgtga tctcttctcg agtcatgtaa ttagttatgt
     7021 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     7081 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     7141 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     7201 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttca ccgcggtgga
     7261 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     7321 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     8041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     8101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     8161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     8281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     9121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtacctctga
     9481 cacatgcagc tcctaacact cacagtgaaa gaaattacag tcgaaacgta tcttaaacca
     9541 tacctgcatc agcaagtaat tcgagcccgt gaatcaccga cttgctgaaa ctaacggtca
     9601 gcaggacttg cggtcgacag gatgagcggt tgtgtcgcat acgagtgcta aattcccctc
     9661 actgttaccc agcgctacct caatagatta gcttataagc acaccttgtt tcacacacct
     9721 ccggagacgg tcacagcttg tacgtctaag aaaccattat tatcatgaca ttaacctata
     9781 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     9841 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     9901 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     9961 cggcatcaga gcagattgta ctgagagtgc accataaatt cccgttttaa gagcttggtg
    10021 agcgctagga gtcactgcca ggtatcgttt gaacacggca ttagtcaggg aagtcataac
    10081 acagtccttt cccgcaattt tctttttcta ttactcttgg cctcctctag tacactctat
    10141 atttttttat gcctcggtaa tgattttcat tttttttttt ccacctagcg gatgactctt
    10201 tttttttctt agcgattggc attatcacat aatgaattat acattatata aagtaatgtg
    10261 atttcttcga agaatatact aaaaaatgag caggcaagat aaacgaaggc aaagatgaca
    10321 gagcagaaag ccctagtaaa gcgtattaca aatgaaacca agattcagat tgcgatctct
    10381 ttaaagggtg gtcccctagc gatagagcac tcgatcttcc cagaaaaaga ggcagaagca
    10441 gtagcagaac aggccacaca atcgcaagtg attaacgtcc acacaggtat agggtttctg
    10501 gaccatatga tacatgctct ggccaagcat tccggctggt cgctaatcgt tgagtgcatt
    10561 ggtgacttac acatagacga ccatcacacc actgaagact gcgggattgc tctcggtcaa
    10621 gcttttaaag aggccctagg ggccgtgcgt ggagtaaaaa ggtttggatc aggatttgcg
    10681 cctttggatg aggcactttc cagagcggtg gtagatcttt cgaacaggcc gtacgcagtt
    10741 gtcgaacttg gtttgcaaag ggagaaagta ggagatctct cttgcgagat gatcccgcat
    10801 tttcttgaaa gctttgcaga ggctagcaga attaccctcc acgttgattg tctgcgaggc
    10861 aagaatgatc atcaccgtag tgagagtgcg ttcaaggctc ttgcggttgc cataagagaa
    10921 gccacctcgc ccaatg
//
