LOCUS       sequence-75626-e       13814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas based plant genome editing and gene regulation; expresses
            dCas9-VP64, gRNA scaffold for insertion of target sequence (OsU3 
            promoter), Bar resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13814)
  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 13814)
  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: pBUN6A11
FEATURES             Location/Qualifiers
     source          1..13814
                     /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"
     promoter        164..543
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     CDS             823..1614
                     /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        1766..1841
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2151..4141
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             4187..4252
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4259..4279
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4304..4327,4328..4330,4331..6817,6818..6820,
                     6821..8404)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             8317..8325
                     /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             8405..8452
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8471..8491
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8507..8527
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8534..8683
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             8690..8710
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8717..8743
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      8791..9043
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(9052..9069)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9052..9068)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9061..9083)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        9268..9945
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             9990..10541
                     /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    10548..10722
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10872..10896
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(11144..11167)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      complement(11175..11202)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(11294..11380)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      11549..11984
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      complement(12099..12687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12179..12198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12777..13586)
                     /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     13500..13519
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     13578..13597
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc taagctcacg tgacggaatt aagcttagta attcatccag
      121 gtcaccaagt tctaggattt tcagaactgc aacttatttt atcaaggaat ctttaaacat
      181 acgaacagat cacttaaagt tcttctgaag caacttaaag ttatcaggca tgcatggatc
      241 ttggaggaat cagatgtgca gtcagggacc atagcacaag acaggcgtct tctactggtg
      301 ctaccagcaa atgctggaag ccgggaacac tgggtacgtt ggaaaccacg tgatgtgaag
      361 aagtaagata aactgtagga gaaaagcatt tcgtagtggg ccatgaagcc tttcaggaca
      421 tgtattgcag tatgggccgg cccattacgc aattggacga caacaaagac tagtattagt
      481 accacctcgg ctatccacat agatcaaagc tgatttaaaa gagttgtgca gatgatccgt
      541 ggcgtgagac caacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
      601 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
      661 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
      721 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
      781 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatggggga agcggtgatc
      841 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
      901 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
      961 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     1021 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     1081 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     1141 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     1201 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     1261 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     1321 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     1381 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     1441 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     1501 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     1561 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta
     1621 gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa gcgttagatg
     1681 cactaagcac ataattgctc acagccaaac tatcaggtca agtctgcttt tattattttt
     1741 aagcgtgcat aataagccgg tctcggtttt agagctagaa atagcaagtt aaaataaggc
     1801 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttttt tcgttttgca
     1861 ttgagttttc tccgtcgcat gtttgcagtt ttattttccg ttttgcattg aaatttctcc
     1921 gtctcatgtt tgcagcgtgt tcaaaaagta cgcagctgta tttcacttat ttacggcgcc
     1981 acattttcat gccgtttgtg ccaactatcc cgagctagtg aatacagctt ggcttcacac
     2041 aacactggtg acccgctgac ctgctcgtac ctcgtaccgt cgtacggcac agcatttgga
     2101 attaaagggt gtgatcgata ctgcttgctg ctaagcttga tatcgaattc ctgcagtgca
     2161 gcgtgacccg gtcgtgcccc tctctagaga taatgagcat tgcatgtcta agttataaaa
     2221 aattaccaca tatttttttt gtcacacttg tttgaagtgc agtttatcta tctttataca
     2281 tatatttaaa ctttactcta cgaataatat aatctatagt actacaataa tatcagtgtt
     2341 ttagagaatc atataaatga acagttagac atggtctaaa ggacaattga gtattttgac
     2401 aacaggactc tacagtttta tctttttagt gtgcatgtgt tctccttttt ttttgcaaat
     2461 agcttcacct atataatact tcatccattt tattagtaca tccatttagg gtttagggtt
     2521 aatggttttt atagactaat ttttttagta catctatttt attctatttt agcctctaaa
     2581 ttaagaaaac taaaactcta ttttagtttt tttatttaat aatttagata taaaatagaa
     2641 taaaataaag tgactaaaaa ttaaacaaat accctttaag aaattaaaaa aactaaggaa
     2701 acatttttct tgtttcgagt agataatgcc agcctgttaa acgccgtcga cgagtctaac
     2761 ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa gcgaagcaga cggcacggca
     2821 tctctgtcgc tgcctctgga cccctctcga gagttccgct ccaccgttgg acttgctccg
     2881 ctgtcggcat ccagaaatgc gtggcggagc ggcagacgtg agccggcacg gcaggcggcc
     2941 tcctcctcct ctcacggcac ggcagctacg ggggattcct ttcccaccgc tccttcgctt
     3001 tcccttcctc gcccgccgta ataaatagac accccctcca caccctcttt ccccaacctc
     3061 gtgttgttcg gagcgcacac acacacaacc agatctcccc caaatccacc cgtcggcacc
     3121 tccgcttcaa ggtacgccgc tcgtcctccc cccccccccc tctctacctt ctctagatcg
     3181 gcgttccggt ccatggttag ggcccggtag ttctacttct gttcatgttt gtgttagatc
     3241 cgtgtttgtg ttagatccgt gctgctagcg ttcgtacacg gatgcgacct gtacgtcaga
     3301 cacgttctga ttgctaactt gccagtgttt ctctttgggg aatcctggga tggctctagc
     3361 cgttccgcag acgggatcga tttcatgatt ttttttgttt cgttgcatag ggtttggttt
     3421 gcccttttcc tttatttcaa tatatgccgt gcacttgttt gtcgggtcat cttttcatgc
     3481 ttttttttgt cttggttgtg atgatgtggt ctggttgggc ggtcgttcta gatcggagta
     3541 gaattctgtt tcaaactacc tggtggattt attaattttg gatctgtatg tgtgtgccat
     3601 acatattcat agttacgaat tgaagatgat ggatggaaat atcgatctag gataggtata
     3661 catgttgatg cgggttttac tgatgcatat acagagatgc tttttgttcg cttggttgtg
     3721 atgatgtggt gtggttgggc ggtcgttcat tcgttctaga tcggagtaga atactgtttc
     3781 aaactacctg gtgtatttat taattttgga actgtatgtg tgtgtcatac atcttcatag
     3841 ttacgagttt aagatggatg gaaatatcga tctaggatag gtatacatgt tgatgtgggt
     3901 tttactgatg catatacatg atggcatatg cagcatctat tcatatgctc taaccttgag
     3961 tacctatcta ttataataaa caagtatgtt ttataattat tttgatcttg atatacttgg
     4021 atgatggcat atgcagcagc tatatgtgga tttttttagc cctgccttca tacgctattt
     4081 atttgcttgg tactgtttct tttgtcgatg ctcaccctgt tgtttggtgt tacttctgca
     4141 gcccggggga tccccaatac ttgtatggcc gcggccgctc tagatggatt acaaggacca
     4201 cgacggggat tacaaggacc acgacattga ttacaaggat gatgatgaca agatggctcc
     4261 gaagaagaag aggaaggttg gcatccacgg ggtgccagct gctgacaaga agtactcgat
     4321 cggcctcgcc attgggacta actctgttgg ctgggccgtg atcaccgacg agtacaaggt
     4381 gccctcaaag aagttcaagg tcctgggcaa caccgatcgg cattccatca agaagaatct
     4441 cattggcgct ctcctgttcg acagcggcga gacggctgag gctacgcggc tcaagcgcac
     4501 cgcccgcagg cggtacacgc gcaggaagaa tcgcatctgc tacctgcagg agattttctc
     4561 caacgagatg gcgaaggttg acgattcttt cttccacagg ctggaggagt cattcctcgt
     4621 ggaggaggat aagaagcacg agcggcatcc aatcttcggc aacattgtcg acgaggttgc
     4681 ctaccacgag aagtacccta cgatctacca tctgcggaag aagctcgtgg actccacaga
     4741 taaggcggac ctccgcctga tctacctcgc tctggcccac atgattaagt tcaggggcca
     4801 tttcctgatc gagggggatc tcaacccgga caatagcgat gttgacaagc tgttcatcca
     4861 gctcgtgcag acgtacaacc agctcttcga ggagaacccc attaatgcgt caggcgtcga
     4921 cgcgaaggct atcctgtccg ctaggctctc gaagtctcgg cgcctcgaga acctgatcgc
     4981 ccagctgccg ggcgagaaga agaacggcct gttcgggaat ctcattgcgc tcagcctggg
     5041 gctcacgccc aacttcaagt cgaatttcga tctcgctgag gacgccaagc tgcagctctc
     5101 caaggacaca tacgacgatg acctggataa cctcctggcc cagatcggcg atcagtacgc
     5161 ggacctgttc ctcgctgcca agaatctgtc ggacgccatc ctcctgtctg atattctcag
     5221 ggtgaacacc gagattacga aggctccgct ctcagcctcc atgatcaagc gctacgacga
     5281 gcaccatcag gatctgaccc tcctgaaggc gctggtcagg cagcagctcc ccgagaagta
     5341 caaggagatc ttcttcgatc agtcgaagaa cggctacgct gggtacattg acggcggggc
     5401 ctctcaggag gagttctaca agttcatcaa gccgattctg gagaagatgg acggcacgga
     5461 ggagctgctg gtgaagctca atcgcgagga cctcctgagg aagcagcgga cattcgataa
     5521 cggcagcatc ccacaccaga ttcatctcgg ggagctgcac gctatcctga ggaggcagga
     5581 ggacttctac cctttcctca aggataaccg cgagaagatc gagaagattc tgactttcag
     5641 gatcccgtac tacgtcggcc cactcgctag gggcaactcc cgcttcgctt ggatgacccg
     5701 caagtcagag gagacgatca cgccgtggaa cttcgaggag gtggtcgaca agggcgctag
     5761 cgctcagtcg ttcatcgaga ggatgacgaa tttcgacaag aacctgccaa atgagaaggt
     5821 gctccctaag cactcgctcc tgtacgagta cttcacagtc tacaacgagc tgactaaggt
     5881 gaagtatgtg accgagggca tgaggaagcc ggctttcctg tctggggagc agaagaaggc
     5941 catcgtggac ctcctgttca agaccaaccg gaaggtcacg gttaagcagc tcaaggagga
     6001 ctacttcaag aagattgagt gcttcgattc ggtcgagatc tctggcgttg aggaccgctt
     6061 caacgcctcc ctggggacct accacgatct cctgaagatc attaaggata aggacttcct
     6121 ggacaacgag gagaatgagg atatcctcga ggacattgtg ctgacactca ctctgttcga
     6181 ggaccgggag atgatcgagg agcgcctgaa gacttacgcc catctcttcg atgacaaggt
     6241 catgaagcag ctcaagagga ggaggtacac cggctggggg aggctgagca ggaagctcat
     6301 caacggcatt cgggacaagc agtccgggaa gacgatcctc gacttcctga agagcgatgg
     6361 cttcgcgaac cgcaatttca tgcagctgat tcacgatgac agcctcacat tcaaggagga
     6421 tatccagaag gctcaggtga gcggccaggg ggactcgctg cacgagcata tcgcgaacct
     6481 cgctggctcg ccagctatca agaaggggat tctgcagacc gtgaaggttg tggacgagct
     6541 ggtgaaggtc atgggcaggc acaagcctga gaacatcgtc attgagatgg cccgggagaa
     6601 tcagaccacg cagaagggcc agaagaactc acgcgagagg atgaagagga tcgaggaggg
     6661 cattaaggag ctggggtccc agatcctcaa ggagcacccg gtggagaaca cgcagctgca
     6721 gaatgagaag ctctacctgt actacctcca gaatggccgc gatatgtatg tggaccagga
     6781 gctggatatt aacaggctca gcgattacga cgtcgatgcc atcgttccac agtcattcct
     6841 gaaggatgac tccattgaca acaaggtcct caccaggtcg gacaagaacc ggggcaagtc
     6901 tgataatgtt ccttcagagg aggtcgttaa gaagatgaag aactactggc gccagctcct
     6961 gaatgccaag ctgatcacgc agcggaagtt cgataacctc acaaaggctg agaggggcgg
     7021 gctctctgag ctggacaagg cgggcttcat caagaggcag ctggtcgaga cacggcagat
     7081 cactaagcac gttgcgcaga ttctcgactc acggatgaac actaagtacg atgagaatga
     7141 caagctgatc cgcgaggtga aggtcatcac cctgaagtca aagctcgtct ccgacttcag
     7201 gaaggatttc cagttctaca aggttcggga gatcaacaat taccaccatg cccatgacgc
     7261 gtacctgaac gcggtggtcg gcacagctct gatcaagaag tacccaaagc tcgagagcga
     7321 gttcgtgtac ggggactaca aggtttacga tgtgaggaag atgatcgcca agtcggagca
     7381 ggagattggc aaggctaccg ccaagtactt cttctactct aacattatga atttcttcaa
     7441 gacagagatc actctggcca atggcgagat ccggaagcgc cccctcatcg agacgaacgg
     7501 cgagacgggg gagatcgtgt gggacaaggg cagggatttc gcgaccgtca ggaaggttct
     7561 ctccatgcca caagtgaata tcgtcaagaa gacagaggtc cagactggcg ggttctctaa
     7621 ggagtcaatt ctgcctaagc ggaacagcga caagctcatc gcccgcaaga aggactggga
     7681 tccgaagaag tacggcgggt tcgacagccc cactgtggcc tactcggtcc tggttgtggc
     7741 gaaggttgag aagggcaagt ccaagaagct caagagcgtg aaggagctgc tggggatcac
     7801 gattatggag cgctccagct tcgagaagaa cccgatcgat ttcctggagg cgaagggcta
     7861 caaggaggtg aagaaggacc tgatcattaa gctccccaag tactcactct tcgagctgga
     7921 gaacggcagg aagcggatgc tggcttccgc tggcgagctg cagaagggga acgagctggc
     7981 tctgccgtcc aagtatgtga acttcctcta cctggcctcc cactacgaga agctcaaggg
     8041 cagccccgag gacaacgagc agaagcagct gttcgtcgag cagcacaagc attacctcga
     8101 cgagatcatt gagcagattt ccgagttctc caagcgcgtg atcctggccg acgcgaatct
     8161 ggataaggtc ctctccgcgt acaacaagca ccgcgacaag ccaatcaggg agcaggctga
     8221 gaatatcatt catctcttca ccctgacgaa cctcggcgcc cctgctgctt tcaagtactt
     8281 cgacacaact atcgatcgca agaggtacac aagcactaag gaggtcctgg acgcgaccct
     8341 catccaccag tcgattaccg gcctctacga gacgcgcatc gacctgtctc agctcggggg
     8401 cgacaagcgg ccagcggcga cgaagaaggc ggggcaggcg aagaagaaga agtccgggtc
     8461 atcaggcagc cctaagaaga agaggaaggt cggcagcagc ggcagcccca agaagaagag
     8521 gaaggtcggg tcggacgccc tcgacgattt cgacctcgat atgctgggct ccgacgccct
     8581 ggacgacttc gacctcgata tgctcgggag cgacgcgctc gacgactttg acctcgatat
     8641 gctcgggtcg gacgctctgg acgattttga cctcgatatg ctggggtcgc ctaagaagaa
     8701 gcggaaggtt ggctcatacc catacgatgt gcccgactac gcctcctgag ctcaggcctc
     8761 catacaagta ttggggatcc gaatttcccc gatcgttcaa acatttggca ataaagtttc
     8821 ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac
     8881 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
     8941 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
     9001 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgggaatt cactggccgt
     9061 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     9121 acatccccct ttcgccaggg gtaccgagct ctagaattcg aggcggtttg cgtattggct
     9181 agagcagctt gccaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga
     9241 tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa tatcgggaaa
     9301 cctcctcgga ttccattgcc cagctatctg tcacttcatc aaaaggacag tagaaaagga
     9361 aggtggcacc tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc
     9421 tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga
     9481 cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat aacatggtgg agcacgacac
     9541 tctcgtctac tccaagaata tcaaagatac agtctcagaa gaccaaaggg ctattgagac
     9601 ttttcaacaa agggtaatat cgggaaacct cctcggattc cattgcccag ctatctgtca
     9661 cttcatcaaa aggacagtag aaaaggaagg tggcacctac aaatgccatc attgcgataa
     9721 aggaaaggct atcgttcaag atgcctctgc cgacagtggt cccaaagatg gacccccacc
     9781 cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg
     9841 atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagacct
     9901 tcctctatat aaggaagttc atttcatttg gagaggacac gctgaaatca ccagtctctc
     9961 tctacaaatc tatctctctc gagtctacca tgagcccaga acgacgcccg gccgacatcc
    10021 gccgtgccac cgaggcggac atgccggcgg tctgcaccat cgtcaaccac tacatcgaga
    10081 caagcacggt caacttccgt accgagccgc aggaaccgca ggagtggacg gacgacctcg
    10141 tccgtctgcg ggagcgctat ccctggctcg tcgccgaggt ggacggcgag gtcgccggca
    10201 tcgcctacgc gggcccctgg aaggcacgca acgcctacga ctggacggcc gagtcgaccg
    10261 tgtacgtctc cccccgccac cagcggacgg gactgggctc cacgctctac acccacctgc
    10321 tgaagtccct ggaggcacag ggcttcaaga gcgtggtcgc tgtcatcggg ctgcccaacg
    10381 acccgagcgt gcgcatgcac gaggcgctcg gatatgcccc ccgcggcatg ctgcgggcgg
    10441 ccggcttcaa gcacgggaac tggcatgacg tgggtttctg gcagctggac ttcagcctgc
    10501 cggtaccgcc ccgtccggtc ctgcccgtca ccgagatttg actcgagttt ctccataata
    10561 atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg ctcatgtgtt
    10621 gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat
    10681 ttctaattcc taaaaccaaa atccagtact aaaatccaga tcccccgaat taattcggcg
    10741 ttaattcagt acattaaaaa cgtccgcaat gtgttattaa gttgtcacta gtcaggttaa
    10801 ctcaattcgg cgttaattca gtacattaaa aacgtccgca atgtgttatt aagttgtcta
    10861 agcgtcaatt tgtttacacc acaatatatc ctgccaccag ccagccaaca gctccccgac
    10921 cggcagctcg gcacaaaatc accactcgat acaggcagcc catcagtccg ggacggcgtc
    10981 agcgggagag ccgttgtaag gcggcagact ttgctcatgt taccgatgct attcggaaga
    11041 acggcaacta agctgccggg tttgaaacac ggatgatctc gcggagggta gcatgttgat
    11101 tgtaacgatg acagagcgtt gctgcctgtg atcacttaag taactaacta acaggaagag
    11161 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
    11221 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
    11281 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
    11341 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
    11401 ctcgcttagt agttagacgt ccccgagatc catgctagac catgaatcca gaagcccgag
    11461 aggttgccgc ctttcgggct ttttcttttt caaaaaaaaa aatttataaa acgatctgtt
    11521 gcggccggcc gccgggttgt gggcaaaggc gctcgacggt gggcaaccgc ttgcggttgt
    11581 ccacgggcgg agccggtgcg cgtagcgcat tgtccacaag ccaagggcga ccaataattg
    11641 atatatatat tcataattga aaagctaatt gaacatacta cttgctgtaa ctacttgccg
    11701 gagcgagggg tgtttgcaag ctgttgatct gaaagggcta ttagcgttct cacgtgcctt
    11761 tttgattagc gatttcacgt gaccttatta gcgatttcac gtactccgat tagcgatttc
    11821 acgtaccctg attagcgatt tcacgtggat agtttttgga gcgggccgga aagccccgtg
    11881 aatcaaggct ttgcggggca ttagcggttt cacgtggata actaccctct atccacaggc
    11941 ttccggggat aaaaaagccc gctcgacggc gggctgttgg atggggatct agcggtaata
    12001 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    12061 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    12121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    12181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    12241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    12301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    12361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    12421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    12481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    12541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    12601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    12661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac cgggtctgac
    12721 gctcagtgga acggggccca atctgaataa tgttacaacc aattaaccaa ttctgattag
    12781 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
    12841 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
    12901 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
    12961 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
    13021 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
    13081 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
    13141 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
    13201 aaccggcgca gggacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
    13261 tctaatacct ggaatgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca
    13321 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
    13381 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
    13441 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta
    13501 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
    13561 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
    13621 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
    13681 gacacgggcc agagctgcag tttgatcccg aggggaaccc tgtggttgac atgcacatac
    13741 aaatggacga acggataaac cttttcacgc ccttttaaat atccgttatt ctaataaacg
    13801 ctcttttctc ttag
//
