LOCUS       sequence-322016-       13499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module for the expression of the human codon optimized Cas9 and 
            kanamycin resistance (NptII)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13499)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 13499)
  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"
COMMENT     Sequence Label: pDGB3 alpha1 Tnos:nptII:Pnos - SF - P35S:hCas9...
FEATURES             Location/Qualifiers
     source          1..13499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             729..1802
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      1868..2062
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(2298..2320)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2406..2546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2461..2480
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2561..2578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2732..3320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2812..2831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3408..4223)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3408..4223)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4131..4150
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    4656..4680
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(4725..4977)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(5017..5811)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5128..5147)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5738..5757
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5845..6024)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        6771..7115
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     7059..7079
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             7189..11292
                     /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             7189..11292
                     /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             7189..11292
                     /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             11305..11325
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      11352..11604
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(11639..11657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    11845..11869
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             join(13170..13499,1..300)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gacatatggg ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat
       61 ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt
      121 gaggttgccg aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag
      181 cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag
      241 ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga
      301 gttaatgagg taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga
      361 gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc
      421 gggtcaactt tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc
      481 aaggcaagac cattaccgag ctgctatctg aatacatcgc gcagctacca gagtaaatga
      541 gcaaatgaat aaatgagtag atgaatttta gcggctaaag gaggcggcat ggaaaatcaa
      601 gaacaaccag gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg cggttggcca
      661 ggcgtaagcg gctgggttgc ctgccggccc tgcaatggca ctggaacccc caagcccgag
      721 gaatcggcgt gagcggtcgc aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg
      781 atgacctggt ggagaagttg aaggccgcgc aggccgccca gcggcaacgc atcgaggcag
      841 aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc
      901 aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc caagggcgac gagcaaccag
      961 attttttcgt tccgatgctc tatgacgtgg gcacccgcga tagtcgcagc atcatggacg
     1021 tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc
     1081 ttccagacgg gcacgtagag gtttccgcag ggccggccgg catggccagt gtgtgggatt
     1141 acgacctggt actgatggcg gtttcccatc taaccgaatc catgaaccga taccgggaag
     1201 ggaagggaga caagcccggc cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct
     1261 gccggcgagc cgatggcgga aagcagaaag acgacctggt agaaacctgc attcggttaa
     1321 acaccacgca cgttgccatg cagcgtacga agaaggccaa gaacggccgc ctggtgacgg
     1381 tatccgaggg tgaagccttg attagccgct acaagatcgt aaagagcgaa accgggcggc
     1441 cggagtacat cgagatcgag ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga
     1501 acccggacgt gctgacggtt caccccgatt actttttgat cgatcccggc atcggccgtt
     1561 ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga
     1621 cgatctacga acgcagtggc agcgccggag agttcaagaa gttctgtttc accgtgcgca
     1681 agctgatcgg gtcaaatgac ctgccggagt acgatttgaa ggaggaggcg gggcaggctg
     1741 gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg cgaagcatcc gccggttcct
     1801 aatgtacgga gcagatgcta gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggac
     1861 tctttcctgt ggatagcacg tacattggga acccaaagcc gtacattggg aaccggaacc
     1921 cgtacattgg gaacccaaag ccgtacattg ggaaccggtc acacatgtaa gtgactgata
     1981 taaaagagaa aaaaggcgat ttttccgcct aaaactcttt aaaacttatt aaaactctta
     2041 aaacccgcct ggcctgtgca taactgtctg gccagcgcac agccgaagag ctgcaaaaag
     2101 cgcctaccct tcggtcgctg cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg
     2161 ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa tctaccaggg cgcggacaag
     2221 ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc
     2281 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg tgacggtcac agcttgtctg
     2341 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     2401 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     2461 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     2521 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     2581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3361 ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc taggtgatta gaaaaactca
     3421 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     3481 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     3541 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     3601 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     3661 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     3721 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgagt
     3781 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     3841 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     3901 tggaatgctg ttttccctgg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     3961 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     4021 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     4081 tcgggcttcc catacaatcg gtagattgtc gcacctgatt gcccgacatt atcgcgagcc
     4141 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct tgagcaagac
     4201 gtttcccgtt gaatatggct cataacagaa cttattattt ccttcctctt ttctacagta
     4261 tttaaagata ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc
     4321 attctaaaac cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta
     4381 taacatagta tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg
     4441 tcatcgttac aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc
     4501 ttagttgccg ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg
     4561 gctctcccgc tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg
     4621 ccgagctgcc ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac
     4681 ataacgaatt cgtctcagga gagcgatcag cttgcatgcc ggtcgatcta gtaacataga
     4741 tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta
     4801 ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca
     4861 ttacatgtta attattacat gcttaacgta attcaacaga aattatatga taatcatggc
     4921 aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg aacgatctgc
     4981 ttgactctag ctagagtccg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     5041 atagaaggct atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     5101 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     5161 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     5221 catgatattc ggcaagcagg cgtcgccgtg ggtcacgacg agatcctcgc cgtcgggcat
     5281 ccgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     5341 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcggtgttt
     5401 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     5461 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     5521 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     5581 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt ggagttcatt
     5641 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     5701 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     5761 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcctcgatc
     5821 gagttgagag tgaatatgag actctaattg gataccgagg ggaatttatg gaacgtcagt
     5881 ggagcatttt tgacaagaaa tatttgctag ctgatagtga ccttaggcga cttttgaacg
     5941 cgcaataatg gtttctgacg tatgtgctta gctcattaaa ctccagaaac ccgcggctca
     6001 gtggctcctt caacgttgcg gttctgtcag ttccaaacgt aaaacggctt gtcccgcgtc
     6061 atcggcgggg gtcataccgt gactccctta attctcatgt atctccttct ttgttcttca
     6121 tccaccattg ttcttcatca atcatttcgc tgtcaggaga ctagagccaa gctgatctcc
     6181 tttgccccgg agatcaccat ggacgacttt ctctatctct acgatctagg aagaaagttc
     6241 gacggagaag gtgacgatac catgttcacc accgataatg agaagattag cctcttcaat
     6301 ttcagaaaga atgctgaccc acagatggtt agagaggcct acgcggcagg tctgatcaag
     6361 acgatctacc cgagtaataa tctccaggag atcaaatacc ttcccaagaa ggttaaagat
     6421 gcagtcaaaa gattcaggac taactgcatc aagaacacag agaaagatat atttctcaag
     6481 atcagaagta ctattccagt atggacgatt caaggcttgc ttcataaacc aaggcaagta
     6541 atagagattg gagtctctaa gaaagtagtt cctactgaat caaaggccat ggagtcaaaa
     6601 attcagatcg aggatctaac agaactcgcc gtgaagactg gcgaacagtt catacagagt
     6661 cttttacgac tcaatgacaa gaagaaaatc ttcgtcaaca tggtggagca cgacactctc
     6721 gtctactcca agaatatcaa agatacagtc tcagaagacc aaagggctat tgagactttt
     6781 caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc
     6841 atcaaaagga cagtagaaaa ggaaggtggc acctacaaat gccatcattg cgataaagga
     6901 aaggctatcg ttcaagatgc ccctgccgac agtggtccca aagatggacc cccacccacg
     6961 aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt
     7021 gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc
     7081 tctatataag gaagttcatt tcatttggag aggactccgg tatttttaca acaattacca
     7141 caacaaaaca aacaacaaac aacattacaa tttactattc tagtcgaaat ggacaagaag
     7201 tactccattg ggctcgatat cggcacaaac agcgtcggct gggccgtcat tacggacgag
     7261 tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca cagcataaag
     7321 aagaacctca ttggcgccct cctgttcgac tccggggaaa cggccgaagc cacgcggctc
     7381 aaaagaacag cacggcgcag atatacccgc agaaagaatc ggatctgcta cctgcaggag
     7441 atctttagta atgagatggc taaggtggat gactctttct tccataggct ggaggagtcc
     7501 tttttggtgg aggaggataa aaagcacgag cgccacccaa tctttggcaa tatcgtggac
     7561 gaggtggcgt accatgaaaa gtacccaacc atatatcatc tgaggaagaa gcttgtagac
     7621 agtactgata aggctgactt gcggttgatc tatctcgcgc tggcgcatat gatcaaattt
     7681 cggggacact tcctcatcga gggggacctg aacccagaca acagcgatgt cgacaaactc
     7741 tttatccaac tggttcagac ttacaatcag cttttcgaag agaacccgat caacgcatcc
     7801 ggagttgacg ccaaagcaat cctgagcgct aggctgtcca aatcccggcg gctcgaaaac
     7861 ctcatcgcac agctccctgg ggagaagaag aacggcctgt ttggtaatct tatcgccctg
     7921 tcactcgggc tgacccccaa ctttaaatct aacttcgacc tggccgaaga tgccaagctt
     7981 caactgagca aagacaccta cgatgatgat ctcgacaatc tgctggccca gatcggcgac
     8041 cagtacgcag accttttttt ggcggcaaag aacctgtcag acgccattct gctgagtgat
     8101 attctgcgag tgaacacgga gatcaccaaa gctccgctga gcgctagtat gatcaagcgc
     8161 tatgatgagc accaccaaga cttgactttg ctgaaggccc ttgtcagaca gcaactgcct
     8221 gagaagtaca aggaaatttt cttcgatcag tctaaaaatg gctacgccgg atacattgac
     8281 ggcggagcaa gccaggagga attttacaaa tttattaagc ccatcttgga aaaaatggac
     8341 ggcaccgagg agctgctggt aaagcttaac agagaagatc tgttgcgcaa acagcgcact
     8401 ttcgacaatg gaagcatccc ccaccagatt cacctgggcg aactgcacgc tatcctcagg
     8461 cggcaagagg atttctaccc ctttttgaaa gataacaggg aaaagattga gaaaatcctc
     8521 acatttcgga taccctacta tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg
     8581 atgactcgca aatcagaaga gactatcact ccctggaact tcgaggaagt cgtggataag
     8641 ggggcctctg cccagtcctt catcgaaagg atgactaact ttgataaaaa tctgcctaac
     8701 gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact tcacagttta taacgagctc
     8761 accaaggtca aatacgtcac agaagggatg agaaagccag cattcctgtc tggagagcag
     8821 aagaaagcta tcgtggacct cctcttcaag acgaaccgga aagttaccgt gaaacagctc
     8881 aaagaagatt atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag cggagtggag
     8941 gatcgcttca acgcatccct gggaacgtat cacgatctcc tgaaaatcat taaagacaag
     9001 gacttcctgg acaatgagga gaacgaggac attcttgagg acattgtcct cacccttacg
     9061 ttgtttgaag atagggagat gattgaagaa cgcttgaaaa cttacgctca tctcttcgac
     9121 gacaaagtca tgaaacagct caagaggcgc cgatatacag gatgggggcg gctgtcaaga
     9181 aaactgatca atgggatccg agacaagcag agtggaaaga caatcctgga ttttcttaag
     9241 tccgatggat ttgccaaccg gaacttcatg cagttgatcc atgatgactc tctcaccttt
     9301 aaggaggaca tccagaaagc acaagtttct ggccaggggg acagtctcca cgagcacatc
     9361 gctaatcttg caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg
     9421 gatgaactcg tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc
     9481 cgagagaacc aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt
     9541 gaagagggta taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc
     9601 cagcttcaga atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg
     9661 gatcaggaac tggacatcaa tcggctctcc gactacgacg tggatcatat cgtgccccag
     9721 tcttttctca aagatgattc tattgataat aaagtgttga caagatccga taaaaataga
     9781 gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg
     9841 cagctgctga acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa
     9901 cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca
     9961 cgccagatca ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat
    10021 gaaaatgaca aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtttca
    10081 gatttcagaa aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg
    10141 catgatgcct acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctt
    10201 gaatctgaat ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag
    10261 tctgagcagg aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat
    10321 tttttcaaga ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa
    10381 acaaacggag aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgg
    10441 aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc
    10501 ttctccaagg aaagtatcct cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa
    10561 gattgggacc ccaagaaata cggcggattc gattctccta cagtcgctta cagtgtactg
    10621 gttgtggcca aagtggagaa agggaagtct aaaaaactca aaagcgtcaa ggaactgctg
    10681 ggcatcacaa tcatggagcg atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg
    10741 aaaggatata aagaggtcaa aaaagacctc atcattaagc ttcccaagta ctctctcttt
    10801 gagcttgaaa acggccggaa acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac
    10861 gagctggcac tgccctctaa atacgttaat ttcttgtatc tggccagcca ctatgaaaag
    10921 ctcaaaggat ctcccgaaga taatgagcag aagcagctgt tcgtggaaca acacaaacac
    10981 taccttgatg agatcatcga gcaaataagc gaattctcca aaagagtgat cctcgccgac
    11041 gctaacctcg ataaggtgct ttctgcttac aataagcaca gggataagcc catcagggag
    11101 caggcagaaa acattatcca cttgtttact ctgaccaact tgggcgcgcc tgcagccttc
    11161 aagtacttcg acaccaccat agacagaaag cggtacacct ctacaaagga ggtcctggac
    11221 gccacactga ttcatcagtc aattacgggg ctctatgaaa caagaatcga cctctctcag
    11281 ctcggtggag acagcagggc tgaccccaag aagaagagga aggtgtgagc ttggaatgga
    11341 tcttcgatcc cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
    11401 ccggtcttgc gacgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
    11461 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
    11521 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
    11581 cggtgtcatc tatgttacta gatcgggaat tgccaagcta attcttgaag acgaaagggc
    11641 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
    11701 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
    11761 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atgggaccga
    11821 ctcgcgctgt catgagacga attctgacag gatatattgg cgggtaaacc taagagaaaa
    11881 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
    11941 atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt tgatccaacc
    12001 cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtcat ctgaaaacga
    12061 catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt
    12121 tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc ggagacatta
    12181 cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca gcaccgacga
    12241 ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca agctgttttc
    12301 cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct
    12361 acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga
    12421 cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga
    12481 gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat
    12541 tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa
    12601 ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc
    12661 ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt
    12721 gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc
    12781 caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc
    12841 cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg
    12901 tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg
    12961 cccgcgcacc tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag
    13021 ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg
    13081 tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga
    13141 gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag
    13201 gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg
    13261 ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc
    13321 gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga
    13381 aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg
    13441 ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttac
//
