载体基本信息 | |||
载体名称 | pBEN1-SGC | 载体抗性 | Kanamycin |
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载体长度 | 7515 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 |
pBEN1-SGC 载体载体图谱
pBEN1-SGC 载体载体序列
LOCUS Exported 7515 bp ds-DNA circular SYN 17-DEC-2018 DEFINITION Bacterial expression vector pBEN1-SGC, complete sequence. ACCESSION GU725055 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7515) AUTHORS Savitsky P, Bray J, Cooper CDO., Marsden BD, Mahajan P, Burgess-Brown NA, Gileadi O. TITLE High-throughput production of human proteins for crystallization: the SGC experience JOURNAL Unpublished REFERENCE 2 (bases 1 to 7515) AUTHORS Gileadi O, Savitsky P, Burgess-Brown NA, Oppermann U, Ng S. TITLE Direct Submission JOURNAL Submitted (08-FEB-2010) SGC, University of Oxford, ORCRB, Roosevelt Drive, Oxford OX3 7DQ, UK REFERENCE 3 (bases 1 to 7515) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..7515 /organism="Bacterial expression vector pBEN1-SGC" /mol_type="other DNA" /db_xref="taxon:747347" CDS 8..307 /codon_start=1 /product="pBEN-SGC tag" /label=pBEN-SGC tag /note="N-terminal tag, includes SET and SBP regions and TEV protease cleavage site" /protein_id="ADE27957.1" /translation="MDPEEASVTSTEETLTPAQEAARTRAANKARKEAELAAATAEQTS DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREPSGGCKLGLGTENLYFQSM" misc_feature 8..136 /note="SET; derived from plasmid pBEn-SBP-SET1A (Stratagene); Region: solubility enhancement tag" CDS 11..136 /codon_start=1 /product="solubility-enhancement tag derived from T7 phage gene 10B" /label=SET1 /translation="DPEEASVTSTEETLTPAQEAARTRAANKARKEAELAAATAEQ" misc_feature 143..274 /note="streptavidin-binding peptide; derived from plasmid pBEn-SBP-SET1A (Stratagene); Region: SBP" CDS 143..253 /codon_start=1 /product="streptavidin-binding peptide" /label=SBP /note="selected from a peptide library; binds streptavidin with nanomolar affinity (Keefe et al., 2001)" /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP" misc_feature 284..304 /note="protein cleaved after Q; Region: TEV protease cleavage site" CDS 284..304 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" promoter 320..765 /gene="sacR" /label=sacB promoter /note="sacB promoter and control region" CDS 766..2187 /codon_start=1 /product="SacB" /label=SacB /note="levansucrase; negative selection marker; prevents growth on media containing 5% sucrose" /protein_id="ADE27955.1" /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" CDS 2295..2312 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 2379..2426 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 2463..2918 /label=F1 origin /note="F1 origin" CDS complement(3011..3826) /codon_start=1 /product="aminoglycoside 3'-phosphotransferase" /label=aminoglycoside 3'-phosphotransferase /note="kanamycin-resistance protein" /protein_id="ADE27954.1" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 3948..4536 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 4722..4864 /label=bom /note="basis of mobility region from pBR322" CDS complement(4966..5157) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5966..7048) /codon_start=1 /product="LacI" /label=LacI /note="lac repressor" /protein_id="ADE27956.1" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(7049..7126) /gene="lacI" /label=lacI promoter regulatory 7435..7478 /regulatory_class="promoter" /note="regulatory region: T7 promoter and Lac operator" promoter 7435..7453 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 7454..7478 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 7493..7515 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
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