| 质粒名称: | pBUN6I11 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 50580 |
| 存储实验室: | Qi-Jun Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pGreen-like binary vector |
| 载体出品公司: | Mullineaux Lab, see http://www.pgreen.ac.uk/ |
| 载体抗性: | 卡那霉素和大观霉素 |
| 载体类型: | CRISPR,植物表达 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dCas9-KRAB,Cas9 |
| 物种来源: | Synthetic |
| 基因突变: | dCas9在DNA断裂中有缺陷；构建了玉米密码子优化的dCas9-KRAB（kruppel-associated-box）载体，用于靶向敲除感兴趣基因。 |
| 启动子 | Ubi1p |
| 标签/融合蛋白: |
3xFLAG（插入件上的N端子）
NLS（NLS N）
NLS（插入件上的C端子）
HA（插入件上的C端子）
|
| 5' 测序引物 | hSpCas9-R1, CGCTCGTGCTTCTTATCCTC |
|
文献引用方法 |
|
| 材料和方法部分: | pBUN6I11 was a gift from Qi-Jun Chen (Addgene plasmid # 50580 ; http://n2t.net/addgene:50580 ; RRID:Addgene_50580) |
| 参考文献部分: | A CRISPR/Cas9 toolkit for multiplex genome editing in plants. Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ. BMC Plant Biol. 2014 Nov 29;14(1):327. 10.1186/s12870-014-0327-y PubMed 25432517 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pBUN6I11 13880 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION CRISPR/Cas based plant genome editing and gene regulation; expresses
dCas9-KRAB, gRNA scaffold for insertion of target sequence (OsU3
promoter), Bar resistance.
ACCESSION .
VERSION .
KEYWORDS pBUN6I11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13880)
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 13880)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..13880
/organism="synthetic DNA construct"
/mol_type="other DNA"
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 4304..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 8564..8749
/codon_start=1
/gene="ZNF10"
/gene_synonym="KOX1"
/product="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
/label=KRAB
/translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
QLTKPDVILRLEKGEEP"
CDS 8756..8776
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8783..8809
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
terminator 8857..9109
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(9118..9135)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(9118..9134)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9127..9149)
/label=M13/pUC Forward
/note="In lacZ gene"
promoter 9334..10011
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 10056..10607
/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 10614..10788
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 10938..10962
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
primer_bind complement(11210..11233)
/label=pENTR-F
/note="pENTR vectors, forward primer"
terminator 11241..11268
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 11360..11446
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin 11615..12050
/label=pSa ori
/note="origin of replication from bacterial plasmid pSa"
rep_origin complement(12165..12753)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(12245..12264)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(12843..13652)
/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 13566..13585
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 13644..13663
/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 ctcggggtcg ccaaagaaga agcggaaggt cggctcaagc gggtcaccaa agaagaagcg
8521 gaaggtcggc tcgatggacg ctaagtccct caccgcttgg agccgcaccc tggtgacgtt
8581 caaggacgtg ttcgtcgatt tcacaaggga ggagtggaag ctcctggaca ctgctcagca
8641 gatcgtttac cggaacgtga tgctggagaa ctacaagaac ctcgtgtccc tcggctacca
8701 gctgacgaag ccggatgtta ttctccgcct ggagaagggc gaggagcccg ggtcccctaa
8761 gaagaagcgg aaggtcggct cctacccata cgatgtccct gactacgcct catgagctca
8821 ggcctccata caagtattgg ggatccgaat ttccccgatc gttcaaacat ttggcaataa
8881 agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg
8941 aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt
9001 tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc
9061 gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg ggaattcact
9121 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
9181 tgcagcacat ccccctttcg ccaggggtac cgagctctag aattcgaggc ggtttgcgta
9241 ttggctagag cagcttgcca acatggtgga gcacgacact ctcgtctact ccaagaatat
9301 caaagataca gtctcagaag accaaagggc tattgagact tttcaacaaa gggtaatatc
9361 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcaaaa ggacagtaga
9421 aaaggaaggt ggcacctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga
9481 tgcctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa
9541 agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgataaca tggtggagca
9601 cgacactctc gtctactcca agaatatcaa agatacagtc tcagaagacc aaagggctat
9661 tgagactttt caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat
9721 ctgtcacttc atcaaaagga cagtagaaaa ggaaggtggc acctacaaat gccatcattg
9781 cgataaagga aaggctatcg ttcaagatgc ctctgccgac agtggtccca aagatggacc
9841 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
9901 ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
9961 agaccttcct ctatataagg aagttcattt catttggaga ggacacgctg aaatcaccag
10021 tctctctcta caaatctatc tctctcgagt ctaccatgag cccagaacga cgcccggccg
10081 acatccgccg tgccaccgag gcggacatgc cggcggtctg caccatcgtc aaccactaca
10141 tcgagacaag cacggtcaac ttccgtaccg agccgcagga accgcaggag tggacggacg
10201 acctcgtccg tctgcgggag cgctatccct ggctcgtcgc cgaggtggac ggcgaggtcg
10261 ccggcatcgc ctacgcgggc ccctggaagg cacgcaacgc ctacgactgg acggccgagt
10321 cgaccgtgta cgtctccccc cgccaccagc ggacgggact gggctccacg ctctacaccc
10381 acctgctgaa gtccctggag gcacagggct tcaagagcgt ggtcgctgtc atcgggctgc
10441 ccaacgaccc gagcgtgcgc atgcacgagg cgctcggata tgccccccgc ggcatgctgc
10501 gggcggccgg cttcaagcac gggaactggc atgacgtggg tttctggcag ctggacttca
10561 gcctgccggt accgccccgt ccggtcctgc ccgtcaccga gatttgactc gagtttctcc
10621 ataataatgt gtgagtagtt cccagataag ggaattaggg ttcctatagg gtttcgctca
10681 tgtgttgagc atataagaaa cccttagtat gtatttgtat ttgtaaaata cttctatcaa
10741 taaaatttct aattcctaaa accaaaatcc agtactaaaa tccagatccc ccgaattaat
10801 tcggcgttaa ttcagtacat taaaaacgtc cgcaatgtgt tattaagttg tcactagtca
10861 ggttaactca attcggcgtt aattcagtac attaaaaacg tccgcaatgt gttattaagt
10921 tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag ccaacagctc
10981 cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc agtccgggac
11041 ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc gatgctattc
11101 ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg agggtagcat
11161 gttgattgta acgatgacag agcgttgctg cctgtgatca cttaagtaac taactaacag
11221 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
11281 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
11341 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
11401 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
11461 cctactctcg cttagtagtt agacgtcccc gagatccatg ctagaccatg aatccagaag
11521 cccgagaggt tgccgccttt cgggcttttt ctttttcaaa aaaaaaaatt tataaaacga
11581 tctgttgcgg ccggccgccg ggttgtgggc aaaggcgctc gacggtgggc aaccgcttgc
11641 ggttgtccac gggcggagcc ggtgcgcgta gcgcattgtc cacaagccaa gggcgaccaa
11701 taattgatat atatattcat aattgaaaag ctaattgaac atactacttg ctgtaactac
11761 ttgccggagc gaggggtgtt tgcaagctgt tgatctgaaa gggctattag cgttctcacg
11821 tgcctttttg attagcgatt tcacgtgacc ttattagcga tttcacgtac tccgattagc
11881 gatttcacgt accctgatta gcgatttcac gtggatagtt tttggagcgg gccggaaagc
11941 cccgtgaatc aaggctttgc ggggcattag cggtttcacg tggataacta ccctctatcc
12001 acaggcttcc ggggataaaa aagcccgctc gacggcgggc tgttggatgg ggatctagcg
12061 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
12121 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
12181 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
12241 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
12301 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
12361 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
12421 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
12481 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
12541 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
12601 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
12661 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
12721 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctaccggg
12781 tctgacgctc agtggaacgg ggcccaatct gaataatgtt acaaccaatt aaccaattct
12841 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
12901 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
12961 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
13021 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
13081 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
13141 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
13201 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
13261 gaatgcaacc ggcgcaggga cactgccagc gcatcaacaa tattttcacc tgaatcagga
13321 tattcttcta atacctggaa tgctgttttt ccggggatcg cagtggtgag taaccatgca
13381 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
13441 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
13501 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
13561 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
13621 ggcctcgacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
13681 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
13741 ttttgagaca cgggccagag ctgcagtttg atcccgaggg gaaccctgtg gttgacatgc
13801 acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gttattctaa
13861 taaacgctct tttctcttag
//
