载体基本信息 | |||
载体名称 | pEU-HSBC | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 6336 bp | 载体类型 | Structural Genomics Vectors |
载体来源 | Vinarov DA, Lytle BL, Peterson FC, Tyler EM, Volkman BF, Markley JL. | 拷贝数 | High copy number |
3'引物 | M13 rev |
pEU-HSBC 载体载体图谱
pEU-HSBC 载体载体序列
LOCUS Exported 6336 bp ds-DNA circular SYN 22-3-2014 DEFINITION Flexi(R) Vector with the sacB gene, for in vitro transcription and translation of an N-terminally 6xHis-tagged protein. ACCESSION . VERSION . KEYWORDS pEU-HSBC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6336) AUTHORS Vinarov DA, Lytle BL, Peterson FC, Tyler EM, Volkman BF, Markley JL. TITLE Cell-free protein production and labeling protocol for NMR-based structural proteomics. JOURNAL Nat. Methods 2004;1:149-53. PUBMED 15782178 REFERENCE 2 (bases 1 to 6336) AUTHORS Center for Eukaryotic Structural Genomics TITLE Direct Submission COMMENT To remove the sacB gene, insert an ORF between the SgfI site and the PmeI site or a downstream site. FEATURES Location/Qualifiers source 1..6336 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" promoter 373..391 /note="SP6 promoter" /note="promoter for bacteriophage SP6 RNA polymerase" CDS 521..523 /codon_start=1 /product="start codon" /note="ATG" /translation="M" CDS 527..544 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 790..2211 /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 ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" promoter 2297..2327 /note="lac UV5 promoter" /note="E. coli lac promoter with an ""up"" mutation" CDS 2381..3040 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(3795..3811) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 3819..3835 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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)." promoter complement(3843..3873) /note="lac promoter" /note="promoter for the E. coli lac operon" rep_origin complement(4197..4785) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4956..5816) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIRYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5817..5921) /gene="bla" /note="AmpR promoter"
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