载体基本信息 | |||
载体名称 | pGEX-2TK | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 4969 bp | 载体类型 | pGEX Vectors (GE Healthcare) |
载体来源 | GE Healthcare | 拷贝数 | High copy number |
pGEX-2TK 载体载体图谱
pGEX-2TK 载体载体序列
LOCUS Exported 4969 bp ds-DNA circular SYN 09-1-2013 DEFINITION Bacterial vector for expressing GST fusions with a thrombin site and a cAMP kinase site. ACCESSION U13851 VERSION . KEYWORDS pGEX-2TK SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4969) AUTHORS GE Healthcare TITLE Direct Submission FEATURES Location/Qualifiers source 1..4969 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" misc_feature 183..211 /note="tac promoter" /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 219..235 /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)." CDS 258..911 /codon_start=1 /gene=" " /product="glutathione S-transferase from Schistosoma japonicum" /note="GST" /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" CDS 918..935 /codon_start=1 /product="thrombin recognition and cleavage site" /note="thrombin site" /translation="LVPRGS" CDS 936..950 /codon_start=1 /product="cAMP-dependent protein kinase recognition site" /note="kinase site" /note=" " /translation="RRASV" misc_feature 951..966 /note="MCS" /note="multiple cloning site" misc_feature 971..981 /note="stop codons" /note="stop codons in all three reading frames" promoter 1272..1376 /gene="bla" /note="AmpR promoter" CDS 1377..2237 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 2408..2996 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 3240..3317 /gene="lacI (mutant)" /note="lacIq promoter" /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 3318..4400 /codon_start=1 /gene="lacI" /product="lac repressor" /note="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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ"
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