载体基本信息 | |||
载体名称 | pFB-LIC-Bse | 载体抗性 | Ampicillin |
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载体长度 | 6816 bp | 载体类型 | Structural Genomics Vectors |
载体来源 | Savitsky P, Bray J, Cooper CD, Marsden BD, Mahajan P, Burgess-Brown | 拷贝数 | High copy number |
pFB-LIC-Bse 载体载体图谱
pFB-LIC-Bse 载体载体序列
LOCUS Exported 6816 bp ds-DNA circular SYN 20-3-2014 DEFINITION Baculovirus vector encoding an N-terminal 6xHis-TEV cassette and ampicillin resistance plus a SacB negative selection marker, for purification of recombinant proteins. ACCESSION EF199842 VERSION . KEYWORDS pFB-LIC-Bse SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6816) AUTHORS Savitsky P, Bray J, Cooper CD, Marsden BD, Mahajan P, Burgess-Brown NA, Gileadi O. TITLE High-throughput production of human proteins for crystallization: the SGC experience. JOURNAL J. Struct. Biol. 2010;172:3-13. PUBMED 20541610 REFERENCE 2 (bases 1 to 6816) AUTHORS Structural Genomics Consortium TITLE Direct Submission COMMENT For ligation-independent cloning (LIC), linearize with BseRI to remove the SacB cassette, and treat with T4 DNA polymerase plus dGTP. FEATURES Location/Qualifiers source 1..6816 /organism="synthetic DNA construct" /lab_host="Insect Cells" /mol_type="other DNA" promoter 1..92 /gene="polh from Autographa californica" /note="polyhedrin promoter" /note="promoter for the baculovirus polyhedrin gene" CDS 148..150 /codon_start=1 /product="start codon" /note="ATG" /translation="M" CDS 154..171 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 196..216 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /note="TEV site" /translation="ENLYFQS" protein_bind 246..279 /bound_moiety="Cre recombinase" /note="loxP" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 294..739 /gene="sacR" /note="sacB promoter" /note="sacB promoter and control region" CDS 740..2161 /codon_start=1 /gene="Bacillus subtilis sacB" /product="secreted levansucrase that renders bacterial growth sensitive to sucrose" /note="SacB" /note="negative selection marker" /protein_id=" " /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" polyA_signal 2403..2537 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" mobile_element 2566..2731 /mobile_element_type="transposon:Tn7" /note="Tn7L" /note="mini-Tn7 element (left end of the Tn7 transposon)" rep_origin 2915..3370 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3397..3501 /gene="bla" /note="AmpR promoter" CDS 3502..4362 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4533..5121 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" mobile_element 5424..5648 /mobile_element_type="transposon:Tn7" /note="Tn7R" /note="mini-Tn7 element (right end of the Tn7 transposon)" CDS complement(5715..6248) /codon_start=1 /gene="aacC1" /allele=" " /product="gentamycin acetyltransferase" /note="GmR" /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(6437..6465) /gene="intI1 (promoter lies within the coding sequence)" /note="Pc promoter" /note="class 1 integron promoter"
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