LOCUS       sequence-75642-e       13407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas based plant genome editing and gene regulation; expresses
            zCas9D10A, gRNA scaffold for insertion of target sequence (AtU6-26 
            promoter), Bar resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13407)
  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 13407)
  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: pBUN501
FEATURES             Location/Qualifiers
     source          1..13407
                     /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        2038..4028
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             4074..4139
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4146..4166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4191..4214,4215..4217,4218..8291)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             8204..8212
                     /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             8292..8339
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8384..8636
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(8645..8662)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8645..8661)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8654..8676)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        8861..9538
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             9583..10134
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    10141..10315
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10465..10489
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10737..10760)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      complement(10768..10795)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(10887..10973)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      11142..11577
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      complement(11692..12280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11772..11791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12370..13179)
                     /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     13093..13112
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     13171..13190
                     /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 agcttgatat cgaattcctg
     2041 cagtgcagcg tgacccggtc gtgcccctct ctagagataa tgagcattgc atgtctaagt
     2101 tataaaaaat taccacatat tttttttgtc acacttgttt gaagtgcagt ttatctatct
     2161 ttatacatat atttaaactt tactctacga ataatataat ctatagtact acaataatat
     2221 cagtgtttta gagaatcata taaatgaaca gttagacatg gtctaaagga caattgagta
     2281 ttttgacaac aggactctac agttttatct ttttagtgtg catgtgttct cctttttttt
     2341 tgcaaatagc ttcacctata taatacttca tccattttat tagtacatcc atttagggtt
     2401 tagggttaat ggtttttata gactaatttt tttagtacat ctattttatt ctattttagc
     2461 ctctaaatta agaaaactaa aactctattt tagttttttt atttaataat ttagatataa
     2521 aatagaataa aataaagtga ctaaaaatta aacaaatacc ctttaagaaa ttaaaaaaac
     2581 taaggaaaca tttttcttgt ttcgagtaga taatgccagc ctgttaaacg ccgtcgacga
     2641 gtctaacgga caccaaccag cgaaccagca gcgtcgcgtc gggccaagcg aagcagacgg
     2701 cacggcatct ctgtcgctgc ctctggaccc ctctcgagag ttccgctcca ccgttggact
     2761 tgctccgctg tcggcatcca gaaatgcgtg gcggagcggc agacgtgagc cggcacggca
     2821 ggcggcctcc tcctcctctc acggcacggc agctacgggg gattcctttc ccaccgctcc
     2881 ttcgctttcc cttcctcgcc cgccgtaata aatagacacc ccctccacac cctctttccc
     2941 caacctcgtg ttgttcggag cgcacacaca cacaaccaga tctcccccaa atccacccgt
     3001 cggcacctcc gcttcaaggt acgccgctcg tcctcccccc ccccccctct ctaccttctc
     3061 tagatcggcg ttccggtcca tggttagggc ccggtagttc tacttctgtt catgtttgtg
     3121 ttagatccgt gtttgtgtta gatccgtgct gctagcgttc gtacacggat gcgacctgta
     3181 cgtcagacac gttctgattg ctaacttgcc agtgtttctc tttggggaat cctgggatgg
     3241 ctctagccgt tccgcagacg ggatcgattt catgattttt tttgtttcgt tgcatagggt
     3301 ttggtttgcc cttttccttt atttcaatat atgccgtgca cttgtttgtc gggtcatctt
     3361 ttcatgcttt tttttgtctt ggttgtgatg atgtggtctg gttgggcggt cgttctagat
     3421 cggagtagaa ttctgtttca aactacctgg tggatttatt aattttggat ctgtatgtgt
     3481 gtgccataca tattcatagt tacgaattga agatgatgga tggaaatatc gatctaggat
     3541 aggtatacat gttgatgcgg gttttactga tgcatataca gagatgcttt ttgttcgctt
     3601 ggttgtgatg atgtggtgtg gttgggcggt cgttcattcg ttctagatcg gagtagaata
     3661 ctgtttcaaa ctacctggtg tatttattaa ttttggaact gtatgtgtgt gtcatacatc
     3721 ttcatagtta cgagtttaag atggatggaa atatcgatct aggataggta tacatgttga
     3781 tgtgggtttt actgatgcat atacatgatg gcatatgcag catctattca tatgctctaa
     3841 ccttgagtac ctatctatta taataaacaa gtatgtttta taattatttt gatcttgata
     3901 tacttggatg atggcatatg cagcagctat atgtggattt ttttagccct gccttcatac
     3961 gctatttatt tgcttggtac tgtttctttt gtcgatgctc accctgttgt ttggtgttac
     4021 ttctgcagcc cgggggatcc ccaatacttg tatggccgcg gccgctctag atggattaca
     4081 aggaccacga cggggattac aaggaccacg acattgatta caaggatgat gatgacaaga
     4141 tggctccgaa gaagaagagg aaggttggca tccacggggt gccagctgct gacaagaagt
     4201 actcgatcgg cctcgccatt gggactaact ctgttggctg ggccgtgatc accgacgagt
     4261 acaaggtgcc ctcaaagaag ttcaaggtcc tgggcaacac cgatcggcat tccatcaaga
     4321 agaatctcat tggcgctctc ctgttcgaca gcggcgagac ggctgaggct acgcggctca
     4381 agcgcaccgc ccgcaggcgg tacacgcgca ggaagaatcg catctgctac ctgcaggaga
     4441 ttttctccaa cgagatggcg aaggttgacg attctttctt ccacaggctg gaggagtcat
     4501 tcctcgtgga ggaggataag aagcacgagc ggcatccaat cttcggcaac attgtcgacg
     4561 aggttgccta ccacgagaag taccctacga tctaccatct gcggaagaag ctcgtggact
     4621 ccacagataa ggcggacctc cgcctgatct acctcgctct ggcccacatg attaagttca
     4681 ggggccattt cctgatcgag ggggatctca acccggacaa tagcgatgtt gacaagctgt
     4741 tcatccagct cgtgcagacg tacaaccagc tcttcgagga gaaccccatt aatgcgtcag
     4801 gcgtcgacgc gaaggctatc ctgtccgcta ggctctcgaa gtctcggcgc ctcgagaacc
     4861 tgatcgccca gctgccgggc gagaagaaga acggcctgtt cgggaatctc attgcgctca
     4921 gcctggggct cacgcccaac ttcaagtcga atttcgatct cgctgaggac gccaagctgc
     4981 agctctccaa ggacacatac gacgatgacc tggataacct cctggcccag atcggcgatc
     5041 agtacgcgga cctgttcctc gctgccaaga atctgtcgga cgccatcctc ctgtctgata
     5101 ttctcagggt gaacaccgag attacgaagg ctccgctctc agcctccatg atcaagcgct
     5161 acgacgagca ccatcaggat ctgaccctcc tgaaggcgct ggtcaggcag cagctccccg
     5221 agaagtacaa ggagatcttc ttcgatcagt cgaagaacgg ctacgctggg tacattgacg
     5281 gcggggcctc tcaggaggag ttctacaagt tcatcaagcc gattctggag aagatggacg
     5341 gcacggagga gctgctggtg aagctcaatc gcgaggacct cctgaggaag cagcggacat
     5401 tcgataacgg cagcatccca caccagattc atctcgggga gctgcacgct atcctgagga
     5461 ggcaggagga cttctaccct ttcctcaagg ataaccgcga gaagatcgag aagattctga
     5521 ctttcaggat cccgtactac gtcggcccac tcgctagggg caactcccgc ttcgcttgga
     5581 tgacccgcaa gtcagaggag acgatcacgc cgtggaactt cgaggaggtg gtcgacaagg
     5641 gcgctagcgc tcagtcgttc atcgagagga tgacgaattt cgacaagaac ctgccaaatg
     5701 agaaggtgct ccctaagcac tcgctcctgt acgagtactt cacagtctac aacgagctga
     5761 ctaaggtgaa gtatgtgacc gagggcatga ggaagccggc tttcctgtct ggggagcaga
     5821 agaaggccat cgtggacctc ctgttcaaga ccaaccggaa ggtcacggtt aagcagctca
     5881 aggaggacta cttcaagaag attgagtgct tcgattcggt cgagatctct ggcgttgagg
     5941 accgcttcaa cgcctccctg gggacctacc acgatctcct gaagatcatt aaggataagg
     6001 acttcctgga caacgaggag aatgaggata tcctcgagga cattgtgctg acactcactc
     6061 tgttcgagga ccgggagatg atcgaggagc gcctgaagac ttacgcccat ctcttcgatg
     6121 acaaggtcat gaagcagctc aagaggagga ggtacaccgg ctgggggagg ctgagcagga
     6181 agctcatcaa cggcattcgg gacaagcagt ccgggaagac gatcctcgac ttcctgaaga
     6241 gcgatggctt cgcgaaccgc aatttcatgc agctgattca cgatgacagc ctcacattca
     6301 aggaggatat ccagaaggct caggtgagcg gccaggggga ctcgctgcac gagcatatcg
     6361 cgaacctcgc tggctcgcca gctatcaaga aggggattct gcagaccgtg aaggttgtgg
     6421 acgagctggt gaaggtcatg ggcaggcaca agcctgagaa catcgtcatt gagatggccc
     6481 gggagaatca gaccacgcag aagggccaga agaactcacg cgagaggatg aagaggatcg
     6541 aggagggcat taaggagctg gggtcccaga tcctcaagga gcacccggtg gagaacacgc
     6601 agctgcagaa tgagaagctc tacctgtact acctccagaa tggccgcgat atgtatgtgg
     6661 accaggagct ggatattaac aggctcagcg attacgacgt cgatcatatc gttccacagt
     6721 cattcctgaa ggatgactcc attgacaaca aggtcctcac caggtcggac aagaaccggg
     6781 gcaagtctga taatgttcct tcagaggagg tcgttaagaa gatgaagaac tactggcgcc
     6841 agctcctgaa tgccaagctg atcacgcagc ggaagttcga taacctcaca aaggctgaga
     6901 ggggcgggct ctctgagctg gacaaggcgg gcttcatcaa gaggcagctg gtcgagacac
     6961 ggcagatcac taagcacgtt gcgcagattc tcgactcacg gatgaacact aagtacgatg
     7021 agaatgacaa gctgatccgc gaggtgaagg tcatcaccct gaagtcaaag ctcgtctccg
     7081 acttcaggaa ggatttccag ttctacaagg ttcgggagat caacaattac caccatgccc
     7141 atgacgcgta cctgaacgcg gtggtcggca cagctctgat caagaagtac ccaaagctcg
     7201 agagcgagtt cgtgtacggg gactacaagg tttacgatgt gaggaagatg atcgccaagt
     7261 cggagcagga gattggcaag gctaccgcca agtacttctt ctactctaac attatgaatt
     7321 tcttcaagac agagatcact ctggccaatg gcgagatccg gaagcgcccc ctcatcgaga
     7381 cgaacggcga gacgggggag atcgtgtggg acaagggcag ggatttcgcg accgtcagga
     7441 aggttctctc catgccacaa gtgaatatcg tcaagaagac agaggtccag actggcgggt
     7501 tctctaagga gtcaattctg cctaagcgga acagcgacaa gctcatcgcc cgcaagaagg
     7561 actgggatcc gaagaagtac ggcgggttcg acagccccac tgtggcctac tcggtcctgg
     7621 ttgtggcgaa ggttgagaag ggcaagtcca agaagctcaa gagcgtgaag gagctgctgg
     7681 ggatcacgat tatggagcgc tccagcttcg agaagaaccc gatcgatttc ctggaggcga
     7741 agggctacaa ggaggtgaag aaggacctga tcattaagct ccccaagtac tcactcttcg
     7801 agctggagaa cggcaggaag cggatgctgg cttccgctgg cgagctgcag aaggggaacg
     7861 agctggctct gccgtccaag tatgtgaact tcctctacct ggcctcccac tacgagaagc
     7921 tcaagggcag ccccgaggac aacgagcaga agcagctgtt cgtcgagcag cacaagcatt
     7981 acctcgacga gatcattgag cagatttccg agttctccaa gcgcgtgatc ctggccgacg
     8041 cgaatctgga taaggtcctc tccgcgtaca acaagcaccg cgacaagcca atcagggagc
     8101 aggctgagaa tatcattcat ctcttcaccc tgacgaacct cggcgcccct gctgctttca
     8161 agtacttcga cacaactatc gatcgcaaga ggtacacaag cactaaggag gtcctggacg
     8221 cgaccctcat ccaccagtcg attaccggcc tctacgagac gcgcatcgac ctgtctcagc
     8281 tcgggggcga caagcggcca gcggcgacga agaaggcggg gcaggcgaag aagaagaagt
     8341 gagctcaggc ctccatacaa gtattgggga tccgaatttc cccgatcgtt caaacatttg
     8401 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     8461 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     8521 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     8581 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga
     8641 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     8701 atcgccttgc agcacatccc cctttcgcca ggggtaccga gctctagaat tcgaggcggt
     8761 ttgcgtattg gctagagcag cttgccaaca tggtggagca cgacactctc gtctactcca
     8821 agaatatcaa agatacagtc tcagaagacc aaagggctat tgagactttt caacaaaggg
     8881 taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcaaaagga
     8941 cagtagaaaa ggaaggtggc acctacaaat gccatcattg cgataaagga aaggctatcg
     9001 ttcaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
     9061 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gataacatgg
     9121 tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaaa
     9181 gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc
     9241 cagctatctg tcacttcatc aaaaggacag tagaaaagga aggtggcacc tacaaatgcc
     9301 atcattgcga taaaggaaag gctatcgttc aagatgcctc tgccgacagt ggtcccaaag
     9361 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
     9421 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
     9481 cttcgcaaga ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctgaaa
     9541 tcaccagtct ctctctacaa atctatctct ctcgagtcta ccatgagccc agaacgacgc
     9601 ccggccgaca tccgccgtgc caccgaggcg gacatgccgg cggtctgcac catcgtcaac
     9661 cactacatcg agacaagcac ggtcaacttc cgtaccgagc cgcaggaacc gcaggagtgg
     9721 acggacgacc tcgtccgtct gcgggagcgc tatccctggc tcgtcgccga ggtggacggc
     9781 gaggtcgccg gcatcgccta cgcgggcccc tggaaggcac gcaacgccta cgactggacg
     9841 gccgagtcga ccgtgtacgt ctccccccgc caccagcgga cgggactggg ctccacgctc
     9901 tacacccacc tgctgaagtc cctggaggca cagggcttca agagcgtggt cgctgtcatc
     9961 gggctgccca acgacccgag cgtgcgcatg cacgaggcgc tcggatatgc cccccgcggc
    10021 atgctgcggg cggccggctt caagcacggg aactggcatg acgtgggttt ctggcagctg
    10081 gacttcagcc tgccggtacc gccccgtccg gtcctgcccg tcaccgagat ttgactcgag
    10141 tttctccata ataatgtgtg agtagttccc agataaggga attagggttc ctatagggtt
    10201 tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt
    10261 ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcccccg
    10321 aattaattcg gcgttaattc agtacattaa aaacgtccgc aatgtgttat taagttgtca
    10381 ctagtcaggt taactcaatt cggcgttaat tcagtacatt aaaaacgtcc gcaatgtgtt
    10441 attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca
    10501 acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca gcccatcagt
    10561 ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca tgttaccgat
    10621 gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat ctcgcggagg
    10681 gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcactt aagtaactaa
    10741 ctaacaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
    10801 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
    10861 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
    10921 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
    10981 gcagttccct actctcgctt agtagttaga cgtccccgag atccatgcta gaccatgaat
    11041 ccagaagccc gagaggttgc cgcctttcgg gctttttctt tttcaaaaaa aaaaatttat
    11101 aaaacgatct gttgcggccg gccgccgggt tgtgggcaaa ggcgctcgac ggtgggcaac
    11161 cgcttgcggt tgtccacggg cggagccggt gcgcgtagcg cattgtccac aagccaaggg
    11221 cgaccaataa ttgatatata tattcataat tgaaaagcta attgaacata ctacttgctg
    11281 taactacttg ccggagcgag gggtgtttgc aagctgttga tctgaaaggg ctattagcgt
    11341 tctcacgtgc ctttttgatt agcgatttca cgtgacctta ttagcgattt cacgtactcc
    11401 gattagcgat ttcacgtacc ctgattagcg atttcacgtg gatagttttt ggagcgggcc
    11461 ggaaagcccc gtgaatcaag gctttgcggg gcattagcgg tttcacgtgg ataactaccc
    11521 tctatccaca ggcttccggg gataaaaaag cccgctcgac ggcgggctgt tggatgggga
    11581 tctagcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
    11641 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    11701 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    11761 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    11821 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    11881 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    11941 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    12001 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    12061 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    12121 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    12181 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    12241 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    12301 taccgggtct gacgctcagt ggaacggggc ccaatctgaa taatgttaca accaattaac
    12361 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
    12421 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
    12481 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
    12541 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
    12601 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
    12661 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
    12721 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
    12781 aggaatcgaa tgcaaccggc gcagggacac tgccagcgca tcaacaatat tttcacctga
    12841 atcaggatat tcttctaata cctggaatgc tgtttttccg gggatcgcag tggtgagtaa
    12901 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
    12961 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
    13021 tttcagaaac aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga
    13081 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
    13141 taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg tattactgtt
    13201 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
    13261 tcagagattt tgagacacgg gccagagctg cagtttgatc ccgaggggaa ccctgtggtt
    13321 gacatgcaca tacaaatgga cgaacggata aaccttttca cgccctttta aatatccgtt
    13381 attctaataa acgctctttt ctcttag
//
