LOCUS       sequence-75660-e       12729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas based plant genome editing and gene regulation; expresses
            3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence
            (AtU6-26 promoter), Hyg resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12729)
  AUTHORS   Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ
  TITLE     A CRISPR/Cas9 toolkit for multiplex genome editing in plants.
  JOURNAL   BMC Plant Biol. 2014 Nov 29;14(1):327.
  PUBMED    25432517
REFERENCE   2  (bases 1 to 12729)
  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: "BMC Plant 
            Biol."; date: "2014-11-29"; volume: "14(1)"; pages: "327"
COMMENT     Sequence Label: pHSN401
FEATURES             Location/Qualifiers
     source          1..12729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             810..1601
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_RNA        1753..1828
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2442..2786
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2969..3034
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3041..3061
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3086..7186
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3086..7186
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             7099..7107
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7187..7234
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      7285..7537
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        7653..8330
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             8398..9423
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    9463..9637
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    9787..9811
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10059..10082)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      complement(10090..10117)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(10209..10295)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      10464..10899
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      complement(11014..11602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11094..11113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11692..12501)
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     12415..12434
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     12493..12512
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc taagctcacg tgacggaatt aagcttcgac ttgccttccg
      121 cacaatacat catttcttct tagctttttt tcttcttctt cgttcataca gttttttttt
      181 gtttatcagc ttacattttc ttgaaccgta gctttcgttt tcttcttttt aactttccat
      241 tcggagtttt tgtatcttgt ttcatagttt gtcccaggat tagaatgatt aggcatcgaa
      301 ccttcaagaa tttgattgaa taaaacatct tcattcttaa gatatgaaga taatcttcaa
      361 aaggcccctg ggaatctgaa agaagagaag caggcccatt tatatgggaa agaacaatag
      421 tatttcttat ataggcccat ttaagttgaa aacaatcttc aaaagtccca catcgcttag
      481 ataagaaaac gaagctgagt ttatatacag ctagagtcga agtagtgatt gggagaccaa
      541 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
      601 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
      661 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
      721 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
      781 gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga
      841 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
      901 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
      961 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     1021 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     1081 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     1141 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     1201 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     1261 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     1321 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     1381 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     1441 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     1501 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     1561 actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct agaaattcgt
     1621 tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac taagcacata
     1681 attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag cgtgcataat
     1741 aagccggtct cggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     1801 acttgaaaaa gtggcaccga gtcggtgctt ttttttgcaa aattttccag atcgatttct
     1861 tcttcctctg ttcttcggcg ttcaatttct ggggttttct cttcgttttc tgtaactgaa
     1921 acctaaaatt tgacctaaaa aaaatctcaa ataatatgat tcagtggttt tgtacttttc
     1981 agttagttga gttttgcagt tccgatgaga taaaccaata agcttgcatg cctgcaggtc
     2041 aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa
     2101 gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc
     2161 cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac
     2221 aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt
     2281 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg
     2341 tcttcaaagc aagtggattg atgtgataac atggtggagc acgacacact tgtctactcc
     2401 aaaaatatca aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg
     2461 gtaatatccg gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag
     2521 atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc
     2581 gttgaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
     2641 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc
     2701 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
     2761 ggaagttcat ttcatttgga gaggacctcg acctcaacac aacatataca aaacaaacga
     2821 atctcaagca atcaagcatt ctacttctat tgcagcaatt taaatcattt cttttaaagc
     2881 aaaagcaatt ttctgaaaat tttcaccatt tacgaacgat actcgagggg gatccccaat
     2941 acttgtatgg ccgcggccgc tctagatgga ttacaaggac cacgacgggg attacaagga
     3001 ccacgacatt gattacaagg atgatgatga caagatggct ccgaagaaga agaggaaggt
     3061 tggcatccac ggggtgccag ctgctgacaa gaagtactcg atcggcctcg atattgggac
     3121 taactctgtt ggctgggccg tgatcaccga cgagtacaag gtgccctcaa agaagttcaa
     3181 ggtcctgggc aacaccgatc ggcattccat caagaagaat ctcattggcg ctctcctgtt
     3241 cgacagcggc gagacggctg aggctacgcg gctcaagcgc accgcccgca ggcggtacac
     3301 gcgcaggaag aatcgcatct gctacctgca ggagattttc tccaacgaga tggcgaaggt
     3361 tgacgattct ttcttccaca ggctggagga gtcattcctc gtggaggagg ataagaagca
     3421 cgagcggcat ccaatcttcg gcaacattgt cgacgaggtt gcctaccacg agaagtaccc
     3481 tacgatctac catctgcgga agaagctcgt ggactccaca gataaggcgg acctccgcct
     3541 gatctacctc gctctggccc acatgattaa gttcaggggc catttcctga tcgaggggga
     3601 tctcaacccg gacaatagcg atgttgacaa gctgttcatc cagctcgtgc agacgtacaa
     3661 ccagctcttc gaggagaacc ccattaatgc gtcaggcgtc gacgcgaagg ctatcctgtc
     3721 cgctaggctc tcgaagtctc ggcgcctcga gaacctgatc gcccagctgc cgggcgagaa
     3781 gaagaacggc ctgttcggga atctcattgc gctcagcctg gggctcacgc ccaacttcaa
     3841 gtcgaatttc gatctcgctg aggacgccaa gctgcagctc tccaaggaca catacgacga
     3901 tgacctggat aacctcctgg cccagatcgg cgatcagtac gcggacctgt tcctcgctgc
     3961 caagaatctg tcggacgcca tcctcctgtc tgatattctc agggtgaaca ccgagattac
     4021 gaaggctccg ctctcagcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac
     4081 cctcctgaag gcgctggtca ggcagcagct ccccgagaag tacaaggaga tcttcttcga
     4141 tcagtcgaag aacggctacg ctgggtacat tgacggcggg gcctctcagg aggagttcta
     4201 caagttcatc aagccgattc tggagaagat ggacggcacg gaggagctgc tggtgaagct
     4261 caatcgcgag gacctcctga ggaagcagcg gacattcgat aacggcagca tcccacacca
     4321 gattcatctc ggggagctgc acgctatcct gaggaggcag gaggacttct accctttcct
     4381 caaggataac cgcgagaaga tcgagaagat tctgactttc aggatcccgt actacgtcgg
     4441 cccactcgct aggggcaact cccgcttcgc ttggatgacc cgcaagtcag aggagacgat
     4501 cacgccgtgg aacttcgagg aggtggtcga caagggcgct agcgctcagt cgttcatcga
     4561 gaggatgacg aatttcgaca agaacctgcc aaatgagaag gtgctcccta agcactcgct
     4621 cctgtacgag tacttcacag tctacaacga gctgactaag gtgaagtatg tgaccgaggg
     4681 catgaggaag ccggctttcc tgtctgggga gcagaagaag gccatcgtgg acctcctgtt
     4741 caagaccaac cggaaggtca cggttaagca gctcaaggag gactacttca agaagattga
     4801 gtgcttcgat tcggtcgaga tctctggcgt tgaggaccgc ttcaacgcct ccctggggac
     4861 ctaccacgat ctcctgaaga tcattaagga taaggacttc ctggacaacg aggagaatga
     4921 ggatatcctc gaggacattg tgctgacact cactctgttc gaggaccggg agatgatcga
     4981 ggagcgcctg aagacttacg cccatctctt cgatgacaag gtcatgaagc agctcaagag
     5041 gaggaggtac accggctggg ggaggctgag caggaagctc atcaacggca ttcgggacaa
     5101 gcagtccggg aagacgatcc tcgacttcct gaagagcgat ggcttcgcga accgcaattt
     5161 catgcagctg attcacgatg acagcctcac attcaaggag gatatccaga aggctcaggt
     5221 gagcggccag ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cgccagctat
     5281 caagaagggg attctgcaga ccgtgaaggt tgtggacgag ctggtgaagg tcatgggcag
     5341 gcacaagcct gagaacatcg tcattgagat ggcccgggag aatcagacca cgcagaaggg
     5401 ccagaagaac tcacgcgaga ggatgaagag gatcgaggag ggcattaagg agctggggtc
     5461 ccagatcctc aaggagcacc cggtggagaa cacgcagctg cagaatgaga agctctacct
     5521 gtactacctc cagaatggcc gcgatatgta tgtggaccag gagctggata ttaacaggct
     5581 cagcgattac gacgtcgatc atatcgttcc acagtcattc ctgaaggatg actccattga
     5641 caacaaggtc ctcaccaggt cggacaagaa ccggggcaag tctgataatg ttccttcaga
     5701 ggaggtcgtt aagaagatga agaactactg gcgccagctc ctgaatgcca agctgatcac
     5761 gcagcggaag ttcgataacc tcacaaaggc tgagaggggc gggctctctg agctggacaa
     5821 ggcgggcttc atcaagaggc agctggtcga gacacggcag atcactaagc acgttgcgca
     5881 gattctcgac tcacggatga acactaagta cgatgagaat gacaagctga tccgcgaggt
     5941 gaaggtcatc accctgaagt caaagctcgt ctccgacttc aggaaggatt tccagttcta
     6001 caaggttcgg gagatcaaca attaccacca tgcccatgac gcgtacctga acgcggtggt
     6061 cggcacagct ctgatcaaga agtacccaaa gctcgagagc gagttcgtgt acggggacta
     6121 caaggtttac gatgtgagga agatgatcgc caagtcggag caggagattg gcaaggctac
     6181 cgccaagtac ttcttctact ctaacattat gaatttcttc aagacagaga tcactctggc
     6241 caatggcgag atccggaagc gccccctcat cgagacgaac ggcgagacgg gggagatcgt
     6301 gtgggacaag ggcagggatt tcgcgaccgt caggaaggtt ctctccatgc cacaagtgaa
     6361 tatcgtcaag aagacagagg tccagactgg cgggttctct aaggagtcaa ttctgcctaa
     6421 gcggaacagc gacaagctca tcgcccgcaa gaaggactgg gatccgaaga agtacggcgg
     6481 gttcgacagc cccactgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa
     6541 gtccaagaag ctcaagagcg tgaaggagct gctggggatc acgattatgg agcgctccag
     6601 cttcgagaag aacccgatcg atttcctgga ggcgaagggc tacaaggagg tgaagaagga
     6661 cctgatcatt aagctcccca agtactcact cttcgagctg gagaacggca ggaagcggat
     6721 gctggcttcc gctggcgagc tgcagaaggg gaacgagctg gctctgccgt ccaagtatgt
     6781 gaacttcctc tacctggcct cccactacga gaagctcaag ggcagccccg aggacaacga
     6841 gcagaagcag ctgttcgtcg agcagcacaa gcattacctc gacgagatca ttgagcagat
     6901 ttccgagttc tccaagcgcg tgatcctggc cgacgcgaat ctggataagg tcctctccgc
     6961 gtacaacaag caccgcgaca agccaatcag ggagcaggct gagaatatca ttcatctctt
     7021 caccctgacg aacctcggcg cccctgctgc tttcaagtac ttcgacacaa ctatcgatcg
     7081 caagaggtac acaagcacta aggaggtcct ggacgcgacc ctcatccacc agtcgattac
     7141 cggcctctac gagacgcgca tcgacctgtc tcagctcggg ggcgacaagc ggccagcggc
     7201 gacgaagaag gcggggcagg cgaagaagaa gaagtgagct caggcctcca tacaagtatt
     7261 ggggatccac tagcgaattt ccccgatcgt tcaaacattt ggcaataaag tttcttaaga
     7321 ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag
     7381 catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga
     7441 gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat
     7501 aaattatcgc gcgcggtgtc atctatgtta ctagatcgga attcgaggcg gtttgcgtat
     7561 tggctagagc agcttgccaa catggtggag cacgacactc tcgtctactc caagaatatc
     7621 aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag ggtaatatcg
     7681 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag gacagtagaa
     7741 aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat
     7801 gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     7861 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgataacat ggtggagcac
     7921 gacactctcg tctactccaa gaatatcaaa gatacagtct cagaagacca aagggctatt
     7981 gagacttttc aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc
     8041 tgtcacttca tcaaaaggac agtagaaaag gaaggtggca cctacaaatg ccatcattgc
     8101 gataaaggaa aggctatcgt tcaagatgcc tctgccgaca gtggtcccaa agatggaccc
     8161 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
     8221 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     8281 gaccttcctc tatataagga agttcatttc atttggagag gacacgctga aatcaccagt
     8341 ctctctctac aaatctatct ctctcgagct ttcgcagatc ccggggggca atgagatatg
     8401 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     8461 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     8521 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     8581 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg
     8641 gagtttagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     8701 gacctgcctg aaaccgaact gcccgctgtt ctacaaccgg tcgcggaggc tatggatgcg
     8761 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     8821 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     8881 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     8941 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     9001 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     9061 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     9121 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccacga
     9181 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     9241 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     9301 gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt
     9361 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaagaaa
     9421 tagagtagat gccgaccgga tctgtcgatc gacaagctcg agtttctcca taataatgtg
     9481 tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat gtgttgagca
     9541 tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta
     9601 attcctaaaa ccaaaatcca gtactaaaat ccagatcccc cgaattaatt cggcgttaat
     9661 tcagtacatt aaaaacgtcc gcaatgtgtt attaagttgt cactagtcag gttaactcaa
     9721 ttcggcgtta attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt
     9781 caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca
     9841 gctcggcaca aaatcaccac tcgatacagg cagcccatca gtccgggacg gcgtcagcgg
     9901 gagagccgtt gtaaggcggc agactttgct catgttaccg atgctattcg gaagaacggc
     9961 aactaagctg ccgggtttga aacacggatg atctcgcgga gggtagcatg ttgattgtaa
    10021 cgatgacaga gcgttgctgc ctgtgatcac ttaagtaact aactaacagg aagagtttgt
    10081 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
    10141 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
    10201 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
    10261 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
    10321 ttagtagtta gacgtccccg agatccatgc tagaccatga atccagaagc ccgagaggtt
    10381 gccgcctttc gggctttttc tttttcaaaa aaaaaaattt ataaaacgat ctgttgcggc
    10441 cggccgccgg gttgtgggca aaggcgctcg acggtgggca accgcttgcg gttgtccacg
    10501 ggcggagccg gtgcgcgtag cgcattgtcc acaagccaag ggcgaccaat aattgatata
    10561 tatattcata attgaaaagc taattgaaca tactacttgc tgtaactact tgccggagcg
    10621 aggggtgttt gcaagctgtt gatctgaaag ggctattagc gttctcacgt gcctttttga
    10681 ttagcgattt cacgtgacct tattagcgat ttcacgtact ccgattagcg atttcacgta
    10741 ccctgattag cgatttcacg tggatagttt ttggagcggg ccggaaagcc ccgtgaatca
    10801 aggctttgcg gggcattagc ggtttcacgt ggataactac cctctatcca caggcttccg
    10861 gggataaaaa agcccgctcg acggcgggct gttggatggg gatctagcgg taatacggtt
    10921 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    10981 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    11041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    11101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    11161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    11221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    11281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    11341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    11401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    11461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    11521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    11581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctaccgggt ctgacgctca
    11641 gtggaacggg gcccaatctg aataatgtta caaccaatta accaattctg attagaaaaa
    11701 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
    11761 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
    11821 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
    11881 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
    11941 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
    12001 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
    12061 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
    12121 gcgcagggac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
    12181 tacctggaat gctgtttttc cggggatcgc agtggtgagt aaccatgcat catcaggagt
    12241 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
    12301 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
    12361 cgcatcgggc ttcccataca agcgatagat tgtcgcacct gattgcccga cattatcgcg
    12421 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgacgt
    12481 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
    12541 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
    12601 gggccagagc tgcagtttga tcccgagggg aaccctgtgg ttgacatgca catacaaatg
    12661 gacgaacgga taaacctttt cacgcccttt taaatatccg ttattctaat aaacgctctt
    12721 ttctcttag
//
