pET-NLS-Cas9-6xHis

价格:1000元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Addgene
载体名称: pET-NLS-Cas9-6xHis
质粒类型: CRISPR/Cas9载体;细菌基因敲除载体
高拷贝/低拷贝: 高拷贝
插入位点: 限制性内切酶,多克隆位点
启动子: lac/T7
载体大小: 9289 bp
5' 测序引物及序列: T7 forward:5’TAATACGACTCACTATAGGG 3’
3' 测序引物及序列: T7 Terminal:5’GCTAGTTATTGCTCAGCGG 3’
载体标签: NLS (N ter); His tag (C-ter)
载体抗性: Ampicillin
克隆菌株: Mach1
宿主细胞(系): 细菌,如大肠杆菌
备注:  
产品目录号: #62934
稳定性: 稳表达
组成型/诱导型: 诱导型
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pET-NLS-Cas9-6xHis 载体图谱



载体简介

载体序列

LOCUS       pET-NLS-Cas9-6xHis	9289 bp 	DNA	SYN
DEFINITION  pET-NLS-Cas9-6xHis
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..9289
                     /organism="pET-NLS-Cas9-6xHis"
                     /mol_type="other DNA"
     misc_feature    2..29
                     /label="lacO"
     misc_feature    47..63
                     /label="T7_transl_en_RBS"
     CDS             71..4219
                     /label="ORF frame 2"
                     /translation="MPKKKRKVMDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLG
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLK
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDHH
                     HHHH*"
     misc_feature    74..94
                     /label="SV40 NLS"
                     /translation="MPKKKRKVMDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLG
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLK
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDHH
                     HHHH*"
     misc_feature    80..94
                     /label="NLS"
                     /translation="MPKKKRKVMDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLG
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLK
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDHH
                     HHHH*"
     misc_feature    4199..4216
                     /label="6xHIS"
                     /translation="MPKKKRKVMDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLG
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLK
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDHH
                     HHHH*"
     misc_feature    complement(4317..4335)
                     /label="T7_Terminal_primer"
                     /translation="MPKKKRKVMDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLG
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLK
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDHH
                     HHHH*"
     gene            complement(4985..5845)
                     /label="Ampicillin"
                     /gene="Ampicillin"
                     /translation="MPKKKRKVMDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLG
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLK
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDHH
                     HHHH*"
     CDS             complement(4985..5845)
                     /label="ORF frame 1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        complement(5888..5916)
                     /label="AmpR_promoter"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     rep_origin      6003..6622
                     /label="pBR322_origin"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    7021..7043
                     /label="pGEX_3_primer"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    complement(7037..7228)
                     /label="ROP"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    complement(7800..8891)
                     /label="lacI"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     CDS             complement(7800..8759)
                     /label="ORF frame 3"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             complement(8632..9132)
                     /label="ORF frame 2"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     gene            complement(8983..9234)
                     /label="tet (887 - 636)"
                     /gene="tet (887 - 636)"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     misc_feature    9166..9185
                     /label="pBRrevBam_primer"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     promoter        9273..2
                     /label="T7_promoter"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
ORIGIN
    1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
   61 gagatatacc atgcccaaga agaagaggaa ggtgatggat aagaaatact caataggctt
  121 agatatcggc acaaatagcg tcggatgggc ggtgatcact gatgaatata aggttccgtc
  181 taaaaagttc aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa atcttatagg
  241 ggctctttta tttgacagtg gagagacagc ggaagcgact cgtctcaaac ggacagctcg
  301 tagaaggtat acacgtcgga agaatcgtat ttgttatcta caggagattt tttcaaatga
  361 gatggcgaaa gtagatgata gtttctttca tcgacttgaa gagtcttttt tggtggaaga
  421 agacaagaag catgaacgtc atcctatttt tggaaatata gtagatgaag ttgcttatca
  481 tgagaaatat ccaactatct atcatctgcg aaaaaaattg gtagattcta ctgataaagc
  541 ggatttgcgc ttaatctatt tggccttagc gcatatgatt aagtttcgtg gtcatttttt
  601 gattgaggga gatttaaatc ctgataatag tgatgtggac aaactattta tccagttggt
  661 acaaacctac aatcaattat ttgaagaaaa ccctattaac gcaagtggag tagatgctaa
  721 agcgattctt tctgcacgat tgagtaaatc aagacgatta gaaaatctca ttgctcagct
  781 ccccggtgag aagaaaaatg gcttatttgg gaatctcatt gctttgtcat tgggtttgac
  841 ccctaatttt aaatcaaatt ttgatttggc agaagatgct aaattacagc tttcaaaaga
  901 tacttacgat gatgatttag ataatttatt ggcgcaaatt ggagatcaat atgctgattt
  961 gtttttggca gctaagaatt tatcagatgc tattttactt tcagatatcc taagagtaaa
 1021 tactgaaata actaaggctc ccctatcagc ttcaatgatt aaacgctacg atgaacatca
 1081 tcaagacttg actcttttaa aagctttagt tcgacaacaa cttccagaaa agtataaaga
 1141 aatctttttt gatcaatcaa aaaacggata tgcaggttat attgatgggg gagctagcca
 1201 agaagaattt tataaattta tcaaaccaat tttagaaaaa atggatggta ctgaggaatt
 1261 attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg acaacggctc
 1321 tattccccat caaattcact tgggtgagct gcatgctatt ttgagaagac aagaagactt
 1381 ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc
 1441 ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc
 1501 tgaagaaaca attaccccat ggaattttga agaagttgtc gataaaggtg cttcagctca
 1561 atcatttatt gaacgcatga caaactttga taaaaatctt ccaaatgaaa aagtactacc
 1621 aaaacatagt ttgctttatg agtattttac ggtttataac gaattgacaa aggtcaaata
 1681 tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga aagccattgt
 1741 tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag aagattattt
 1801 caaaaaaata gaatgttttg atagtgttga aatttcagga gttgaagata gatttaatgc
 1861 ttcattaggt acctaccatg atttgctaaa aattattaaa gataaagatt ttttggataa
 1921 tgaagaaaat gaagatatct tagaggatat tgttttaaca ttgaccttat ttgaagatag
 1981 ggagatgatt gaggaaagac ttaaaacata tgctcacctc tttgatgata aggtgatgaa
 2041 acagcttaaa cgtcgccgtt atactggttg gggacgtttg tctcgaaaat tgattaatgg
 2101 tattagggat aagcaatctg gcaaaacaat attagatttt ttgaaatcag atggttttgc
 2161 caatcgcaat tttatgcagc tgatccatga tgatagtttg acatttaaag aagacattca
 2221 aaaagcacaa gtgtctggac aaggcgatag tttacatgaa catattgcaa atttagctgg
 2281 tagccctgct attaaaaaag gtattttaca gactgtaaaa gttgttgatg aattggtcaa
 2341 agtaatgggg cggcataagc cagaaaatat cgttattgaa atggcacgtg aaaatcagac
 2401 aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa
 2461 agaattagga agtcagattc ttaaagagca tcctgttgaa aatactcaat tgcaaaatga
 2521 aaagctctat ctctattatc tccaaaatgg aagagacatg tatgtggacc aagaattaga
 2581 tattaatcgt ttaagtgatt atgatgtcga tcacattgtt ccacaaagtt tccttaaaga
 2641 cgattcaata gacaataagg tcttaacgcg ttctgataaa aatcgtggta aatcggataa
 2701 cgttccaagt gaagaagtag tcaaaaagat gaaaaactat tggagacaac ttctaaacgc
 2761 caagttaatc actcaacgta agtttgataa tttaacgaaa gctgaacgtg gaggtttgag
 2821 tgaacttgat aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa
 2881 gcatgtggca caaattttgg atagtcgcat gaatactaaa tacgatgaaa atgataaact
 2941 tattcgagag gttaaagtga ttaccttaaa atctaaatta gtttctgact tccgaaaaga
 3001 tttccaattc tataaagtac gtgagattaa caattaccat catgcccatg atgcgtatct
 3061 aaatgccgtc gttggaactg ctttgattaa gaaatatcca aaacttgaat cggagtttgt
 3121 ctatggtgat tataaagttt atgatgttcg taaaatgatt gctaagtctg agcaagaaat
 3181 aggcaaagca accgcaaaat atttctttta ctctaatatc atgaacttct tcaaaacaga
 3241 aattacactt gcaaatggag agattcgcaa acgccctcta atcgaaacta atggggaaac
 3301 tggagaaatt gtctgggata aagggcgaga ttttgccaca gtgcgcaaag tattgtccat
 3361 gccccaagtc aatattgtca agaaaacaga agtacagaca ggcggattct ccaaggagtc
 3421 aattttacca aaaagaaatt cggacaagct tattgctcgt aaaaaagact gggatccaaa
 3481 aaaatatggt ggttttgata gtccaacggt agcttattca gtcctagtgg ttgctaaggt
 3541 ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag ttactaggga tcacaattat
 3601 ggaaagaagt tcctttgaaa aaaatccgat tgacttttta gaagctaaag gatataagga
 3661 agttaaaaaa gacttaatca ttaaactacc taaatatagt ctttttgagt tagaaaacgg
 3721 tcgtaaacgg atgctggcta gtgccggaga attacaaaaa ggaaatgagc tggctctgcc
 3781 aagcaaatat gtgaattttt tatatttagc tagtcattat gaaaagttga agggtagtcc
 3841 agaagataac gaacaaaaac aattgtttgt Tgagcagcat aagcattatt tagatgagat
 3901 tattgagcaa atcagtgaat tttctaagcg tgttatttta gcagatgcca atttagataa
 3961 agttcttagt gcatataaca aacatagaga caaaccaata cgtgaacaag cagaaaatat
 4021 tattcattta tttacgttga cgaatcttgg agctcccgct gcttttaaat attttgatac
 4081 aacaattgat cgtaaacgat atacgtctac aaaagaagtt ttagatgcca ctcttatcca
 4141 tcaatccatc actggtcttt atgaaacacg cattgatttg agtcagctag gaggtgacca
 4201 tcaccaccac catcactaat gactcgagtc tggtaaagaa accgctgctg cgaaatttga
 4261 acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc
 4321 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
 4381 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
 4441 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
 4501 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
 4561 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
 4621 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
 4681 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
 4741 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
 4801 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
 4861 tttacaattt ctggcggcac gatggcatga gattatcaaa aaggatcttc acctagatcc
 4921 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
 4981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
 5041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
 5101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
 5161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
 5221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
 5281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
 5341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
 5401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
 5461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
 5521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
 5581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
 5641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
 5701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
 5761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
 5821 cgacacggaa atgttgaata ctcatactct tcctttttca atcatgattg aagcatttat
 5881 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
 5941 ggtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
 6001 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
 6061 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
 6121 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
 6181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
 6241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
 6301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
 6361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
 6421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
 6481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
 6541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
 6601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
 6661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
 6721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
 6781 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
 6841 catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
 6901 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
 6961 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
 7021 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
 7081 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
 7141 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
 7201 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
 7261 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
 7321 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
 7381 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
 7441 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
 7501 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
 7561 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
 7621 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
 7681 tcatgctagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
 7741 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
 7801 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
 7861 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
 7921 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
 7981 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
 8041 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
 8101 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
 8161 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
 8221 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
 8281 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
 8341 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
 8401 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
 8461 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
 8521 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
 8581 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
 8641 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
 8701 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
 8761 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
 8821 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
 8881 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
 8941 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
 9001 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
 9061 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
 9121 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
 9181 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
 9241 gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactata
//

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