载体基本信息 | |||
载体名称 | pET-32b(+) | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 5899 bp | 载体类型 | pET & Duet Vectors (Novagen) |
载体来源 | Novagen (EMD Millipore) | 拷贝数 | High copy number |
pET-32b(+) 载体载体图谱
pET-32b(+) 载体载体序列
LOCUS Exported 5899 bp ds-DNA circular SYN 02-12-2013 DEFINITION Bacterial vector for expressing thioredoxin fusion proteins with an enterokinase site. For other reading frames, use pET-32a(+) or pET-32c(+). ACCESSION . VERSION . KEYWORDS pET-32b(+) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5899) AUTHORS Novagen (EMD Millipore) TITLE Direct Submission FEATURES Location/Qualifiers source 1..5899 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" terminator 26..73 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(140..157) /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" misc_feature 158..216 /note="MCS" /note="multiple cloning site" CDS complement(218..232) /codon_start=1 /product="enterokinase recognition and cleavage site" /note="enterokinase site" /translation="DDDDK" CDS complement(248..292) /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /note="S-Tag" /translation="KETAAAKFERQHMDS" CDS complement(299..316) /codon_start=1 /product="thrombin recognition and cleavage site" /note="thrombin site" /translation="LVPRGS" CDS complement(326..343) /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS complement(365..691) /codon_start=1 /gene="trxA" /product="E. coli thioredoxin" /note="TrxA" /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL DANLA" RBS 700..705 /note="ribosome binding site" protein_bind 736..760 /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(761..779) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" promoter 1092..1169 /gene="lacI" /note="lacI promoter" /note=" " CDS 1170..2252 /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" CDS 3061..3252 /codon_start=1 /gene="rop" /product="Rop protein" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" rep_origin complement(3682..4270) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4441..5301) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5302..5406) /gene="bla" /note="AmpR promoter" rep_origin complement(5433..5888) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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