LOCUS       sequence-167042-       14556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate Level 2 construct encoding Cas9-YFP and 2 
            urease-targeting gRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14556)
  AUTHORS   Hopes A, Nekrasov V, Kamoun S, Mock T
  TITLE     Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira
            pseudonana.
  JOURNAL   Plant Methods. 2016 Nov 24;12:49. doi: 10.1186/s13007-016-0148-0. 
            eCollection 2016.
  PUBMED    27904648
REFERENCE   2  (bases 1 to 14556)
  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: "Plant 
            Methods."; date: "2016-11-24"; volume: "12:49. doi"; pages: " 
            10.1186/s13007-016-0148-0. eCollection 2016"
COMMENT     Sequence Label: pAGM4723:TpCC_Urease
FEATURES             Location/Qualifiers
     source          1..14556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    36..60
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             340..969
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1403..2470
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      2536..2730
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        2938..3042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3043..3837
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      4008..4596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4497..4516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4732..4756
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     4846..4862
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4847..4863
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5847..6416
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     primer_bind     complement(6934..6950)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6934..6950)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6991..7011)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6991..7009)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             8017..12120
                     /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             8017..12120
                     /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             8017..12120
                     /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             12133..12153
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(12154..12156,12157..12159,12160..12873)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(12199..12220)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(12460..12479)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     12807..12828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_RNA        13872..13947
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        14447..14522
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gggacacgaa gtgatccgtt taaactatca gtgtttgaca ggatatattg gcgggtaaac
       61 ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta
      121 tccgttcgtc catttgtatg tgccagccgt gcggctgcat gaaatcctgg ccggtttgtc
      181 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
      241 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
      301 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
      361 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
      421 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
      481 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
      541 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
      601 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
      661 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
      721 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
      781 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
      841 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
      901 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
      961 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     1021 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     1081 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     1141 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
     1201 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     1261 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     1321 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc
     1381 aagcccgagg aatcggcgtg acggtcgcaa accatccggc ccggtacaaa tcggcgcggc
     1441 gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat
     1501 cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga
     1561 atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga
     1621 gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat
     1681 catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg
     1741 ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt
     1801 gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata
     1861 ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact
     1921 caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat
     1981 tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct
     2041 ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac
     2101 cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga
     2161 aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat
     2221 cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt
     2281 gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac
     2341 cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg
     2401 gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc
     2461 cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg
     2521 aaaaagcttc tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa
     2581 ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt
     2641 gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa
     2701 aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaacagct
     2761 gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc
     2821 tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg
     2881 cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca catcaaggct ccgagtgcgc
     2941 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3001 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatggctaa aatgagaata
     3061 tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg
     3121 tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg
     3181 gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg
     3241 ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc
     3301 aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa
     3361 agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca ctccatcgac
     3421 atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt ggattactta
     3481 ctgaataacg atctggccga tgtggattgc gaaaactggg aagaggacac tccatttaaa
     3541 gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga acttgtcttt
     3601 tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt aagtggcttt
     3661 attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt ctgcgtccgg
     3721 tcgctcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg
     3781 atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt gttttagctg
     3841 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3901 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3961 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4021 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4081 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4141 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4201 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4261 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4321 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4381 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4441 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4501 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4561 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4621 cggttcctgc tcggatctgt tggaccggac agtagtcatg gttgatgggc tgcctgtatc
     4681 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat
     4741 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
     4801 aatgtactgg ggttgaacac tctgtgccga attcggatcc ggagctcgag gtcgacggta
     4861 tcgataagct tgcgcttttt ccgagaactc cccataagtc aacggctcca atcaagaatg
     4921 tatccgacaa cggcgagcat agcaacacgt ccgtctttgg agtagaatca tcatgttgtg
     4981 gatgaataca cagatgaatg acattaaaag catgaacatg ttagagagta ggaggtagag
     5041 attgatatgg tagcattgcg atgtttgttt ttggtcagca tatgatgagt ggataccaat
     5101 atgatgaaag ttgaatctcg cgtttgagct cagcggtacg ttattgatcg aaagtagcct
     5161 gatcaaaatc cttggagagt acaagaggat caaagaatcc agtgggggcg ataactccaa
     5221 gctcgttctc aaagaggcaa tggaggtaga aactcatccc agttgagaag aagtgaaggc
     5281 agtggcggtg gcgaaagcag aggcaacgag gacagacttc ctgtgggttg atgcaacgaa
     5341 tatttccaga aggagaagtt tagagagttg aaccgctacc tacaatgaca aagtatcgta
     5401 tcgattttga tgttggttgg ttatgaattc aaactgtaag ttggattgtg agaagatcag
     5461 aagttgaacg aacacatctt tccgatcatt cacctccaca ctgcaacaac acggtacttc
     5521 ttccgcggca gatctctgtc gccattctct tgtcctgttg ttggctgtga gacgaggaaa
     5581 gcaacgacaa gtttcacaaa agggagttcc tttaacgaga tatgtttttt ataaagagtc
     5641 ccaatagaaa gacaaattga ttcctccgtg caaacgcgca aataaacacc acgtccatta
     5701 tatccatatc tttcagagta tccaacaagt gttgaaggac aggtagttga agtaacgtat
     5761 cttccccctc gactggatcc atcaacaagg cgaacaaatc cattcaacct ctcataaatt
     5821 atctgattta ccaaaccgat accaaaatga ccactcttga cgacacggct taccggtacc
     5881 gcaccagtgt cccgggggac gccgaggcca tcgaggcact ggatgggtcc ttcaccaccg
     5941 acaccgtatt ccgcgtcacc gccaccgggg acggcttcac cctgcgggag gtgccggtgg
     6001 acccgcccct gaccaaggtg ttccccgacg acgaatcgga cgacgaatcg gacgccgggg
     6061 aggacggcga cccggactcc cggacgttcg tcgcgtacgg ggacgacggc gacctggcgg
     6121 gcttcgtggt cgtctcgtac tccggctgga accgccggct gaccgtcgag gacatcgagg
     6181 tcgccccgga gcaccggggg cacggggtcg ggcgcgcgtt gatggggctc gcgacggagt
     6241 tcgcccgcga gcggggcgcc gggcacctct ggctggaggt caccaacgtc aacgcaccgg
     6301 cgatccacgc gtaccggcgg atggggttca ccctctgcgg cctggacacc gccctgtacg
     6361 acggcaccgc ctcggacggc gagcaggcgc tctacatgag catgccctgc ccctgagcgc
     6421 ggccgcatac tggattggtg aatcaatgag ccgtagcaca atggttacat tcggctagct
     6481 aagatccaat ggcaaggacc aagtgctgga acttgttttg ctttagcaga tcttagcgtg
     6541 agaggtattt gtcctctgtc aggagtagat agtagatgtt ctttttaaac taaaatgcta
     6601 actgttccga attcctcatc gcagctaatc cgtacatcaa aagacaaaat gctaggtatg
     6661 tgtactacat ctcctgttgc tagataagac atatgatagg aaacacacca tcaatagtca
     6721 ttgtagcttt acttatacta cgcatttgca ctttcccctg agtggcagag gcgcattgag
     6781 aaaatcgatc tcaacatagt ttatgtagca tcccctagat ccattacttt aagtctcctt
     6841 cgtctttggt gtaggcatgt tggacacaac gaggtaaaac acaacacaaa caatgtgtcc
     6901 agcaaagtag tagctgctcc agttctcccg ggggatccac tagttctaga gcggccggcc
     6961 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg cgcgctgcaa
     7021 gaattcaagc ttggagagct tgcgcttttt ccgagaactc cccataagtc aacggctcca
     7081 atcaagaatg tatccgacaa cggcgagcat agcaacacgt ccgtctttgg agtagaatca
     7141 tcatgttgtg gatgaataca cagatgaatg acattaaaag catgaacatg ttagagagta
     7201 ggaggtagag attgatatgg tagcattgcg atgtttgttt ttggtcagca tatgatgagt
     7261 ggataccaat atgatgaaag ttgaatctcg cgtttgagct cagcggtacg ttattgatcg
     7321 aaagtagcct gatcaaaatc cttggagagt acaagaggat caaagaatcc agtgggggcg
     7381 ataactccaa gctcgttctc aaagaggcaa tggaggtaga aactcatccc agttgagaag
     7441 aagtgaaggc agtggcggtg gcgaaagcag aggcaacgag gacagacttc ctgtgggttg
     7501 atgcaacgaa tatttccaga aggagaagtt tagagagttg aaccgctacc tacaatgaca
     7561 aagtatcgta tcgattttga tgttggttgg ttatgaattc aaactgtaag ttggattgtg
     7621 agaagatcag aagttgaacg aacacatctt tccgatcatt cacctccaca ctgcaacaac
     7681 acggtacttc ttccgcggca gatctctgtc gccattctct tgtcctgttg ttggctgtga
     7741 gacgaggaaa gcaacgacaa gtttcacaaa agggagttcc tttaacgaga tatgtttttt
     7801 ataaagagtc ccaatagaaa gacaaattga ttcctccgtg caaacgcgca aataaacacc
     7861 acgtccatta tatccatatc tttcagagta tccaacaagt gttgaaggac aggtagttga
     7921 agtaacgtat cttccccctc gactggatcc atcaacaagg cgaacaaatc cattcaacct
     7981 ctcataaatt atctgattta ccaaaccaat accaaaatgg acaagaagta ctccattggg
     8041 ctcgatatcg gcacaaacag cgtcggctgg gccgtcatta cggacgagta caaggtgccg
     8101 agcaaaaaat tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt
     8161 ggcgccctcc tgttcgactc cggggagacg gccgaagcca cgcggctcaa aagaacagca
     8221 cggcgcagat atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat
     8281 gagatggcta aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag
     8341 gaggataaaa agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac
     8401 catgaaaagt acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag
     8461 gctgacttgc ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc
     8521 ctcatcgagg gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg
     8581 gttcagactt acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc
     8641 aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag
     8701 ctccctgggg agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg
     8761 acccccaact ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa
     8821 gacacctacg atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac
     8881 ctttttttgg cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg
     8941 aacacggaga tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac
     9001 caccaagact tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag
     9061 gaaattttct tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc
     9121 caggaggaat tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag
     9181 ctgctggtaa agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga
     9241 agcatccccc accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat
     9301 ttctacccct ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata
     9361 ccctactatg taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa
     9421 tcagaagaga ctatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc
     9481 cagtccttca tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt
     9541 cctaaacact ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa
     9601 tacgtcacag aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc
     9661 gtggacctcc tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaagattat
     9721 ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac
     9781 gcatccctgg gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac
     9841 aatgaggaga acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat
     9901 agggagatga ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg
     9961 aaacagctca agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat
    10021 gggatccgag acaagcagag tggaaagaca atcctggatt ttcttaagtc cgatggattt
    10081 gccaaccgga acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc
    10141 cagaaagcac aagtttctgg ccagggggac agtctccacg agcacatcgc taatcttgca
    10201 ggtagcccag ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc
    10261 aaagtaatgg gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa
    10321 actacccaga agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata
    10381 aaagaactgg ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat
    10441 gagaagctct acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg
    10501 gacatcaatc ggctctccga ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa
    10561 gatgattcta ttgataataa agtgttgaca agatccgata aaaatagagg gaagagtgat
    10621 aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac
    10681 gccaaactga tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg
    10741 tctgagttgg ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc
    10801 aagcacgtgg cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa
    10861 ctgattcgag aggtgaaagt tattactctg aagtctaagc tggtttcaga tttcagaaag
    10921 gactttcagt tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac
    10981 ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt
    11041 gtttacggag actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa
    11101 ataggcaagg ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc
    11161 gagattacac tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa
    11221 acaggagaaa tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc
    11281 atgccgcagg tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa
    11341 agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc
    11401 aagaaatacg gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa
    11461 gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc
    11521 atggagcgat caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa
    11581 gaggtcaaaa aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac
    11641 ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg
    11701 ccctctaaat acgttaattt cttgtatctg gccagccact atgaaaagct caaaggatct
    11761 cccgaagata atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag
    11821 atcatcgagc aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat
    11881 aaggtgcttt ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac
    11941 attatccact tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac
    12001 accaccatag acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt
    12061 catcagtcaa ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac
    12121 agcagggctg accccaagaa gaagaggaag gtgatggtga gcaagggcga ggagctgttc
    12181 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
    12241 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
    12301 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
    12361 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
    12421 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
    12481 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
    12541 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
    12601 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
    12661 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
    12721 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
    12781 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
    12841 atcactctcg gcatggacga gctgtacaag tgagcttata ctggattggt gaatcaatga
    12901 gccgtagcac aatggttaca ttcggctagc taagatccaa tggcaaggac caagtgctgg
    12961 aacttgtttt gctttagcag atcttagcgt gagaggtatt tgtcctctgt caggagtaga
    13021 tagtagatgt tctttttaaa ctaaaatgct aactgttccg aattcctcat cgcagctaat
    13081 ccgtacatca aaagacaaaa tgctaggtat gtgtactaca tctcctgttg ctagataaga
    13141 catatgatag gaaacacacc atcaatagtc attgtagctt tacttatact acgcatttgc
    13201 actttcccct gagtggcaga ggcgcattga gaaaatcgat ctcaacatag tttatgtagc
    13261 atcccctaga tccattactt taagtctcct tcgtctttgg tgtaggcatg ttggacacaa
    13321 cgaggtaaaa cacaacacaa acaatgtgtc cagcaaagta gtagctgctc cagttctccg
    13381 ctactagaat tcgagctcgg agcttcatca agagagcaac caacaaagaa gaggcgaaca
    13441 cggtgcacag caccgtctgc atacttccga tttgcaattc acaacatatg atacatcatc
    13501 catctctctg agaaaactgg gcactgttag aatctgggat ttatcaacag ccaccaacta
    13561 tctactcaac gcaattgggg cacttgataa tgtgacaatc ctccttgctc ctttccttgt
    13621 cgcctcaaac aacgtcggga actgcacaga aacatcgcta cttattcacg cctcacctct
    13681 ctttcctcct ttactcactt attcgcgcct cacctctctt tcctccttta ctcaagtcct
    13741 ctgcttgccg ggtttattct tccgtttgtg ttgttgcggt ctgataggat ttgtttggag
    13801 acgaaccttt ttttttgatt gctttataaa ttaacgtttt gccgaaattg tgtcgtaatc
    13861 aagtattgcc ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
    13921 cttgaaaaag tggcaccgag tcggtgcttt tttcgcttta cgaattccca tggggagctt
    13981 catcaagaga gcaaccaaca aagaagaggc gaacacggtg cacagcaccg tctgcatact
    14041 tccgatttgc aattcacaac atatgataca tcatccatct ctctgagaaa actgggcact
    14101 gttagaatct gggatttatc aacagccacc aactatctac tcaacgcaat tggggcactt
    14161 gataatgtga caatcctcct tgctcctttc cttgtcgcct caaacaacgt cgggaactgc
    14221 acagaaacat cgctacttat tcacgcctca cctctctttc ctcctttact cacttattcg
    14281 cgcctcacct ctctttcctc ctttactcaa gtcctctgct tgccgggttt attcttccgt
    14341 ttgtgttgtt gcggtctgat aggatttgtt tggagacgaa cctttttttt tgattgcttt
    14401 ataaattaac gttttgccga aattgtgttt ccgatctaat gtccatgttt tagagctaga
    14461 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
    14521 gcttttttcg ctcagagagg atgcacatgt gaccga
//
