载体基本信息 | |||
载体名称 | pMCSG25 | 载体抗性 | Ampicillin |
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载体长度 | 5664 bp | 载体类型 | Structural Genomics Vectors |
载体来源 | Osipiuk J, Mulligan R, Bargassa M, Hamilton JE, Cunningham MA, | 拷贝数 | High copy number |
pMCSG25 载体载体图谱
pMCSG25 载体载体序列
LOCUS Exported 5664 bp ds-DNA circular SYN 10-2-2014 DEFINITION Bacterial expression vector encoding an aspartic endopeptidase fused to a 6xHis-TEV leader. ACCESSION . VERSION . KEYWORDS pMCSG25 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5664) AUTHORS Osipiuk J, Mulligan R, Bargassa M, Hamilton JE, Cunningham MA, Joachimiak A. TITLE Characterization of member of DUF1888 protein family, self-cleaving and self-assembling endopeptidase. JOURNAL J. Biol. Chem. 2012;287:19452-61. PUBMED 22493430 REFERENCE 2 (bases 1 to 5664) AUTHORS Midwest Center for Structural Genomics TITLE Direct Submission COMMENT For ligation-independent cloning (LIC), linearize with SspI and treat with T4 DNA polymerase plus dGTP. FEATURES Location/Qualifiers source 1..5664 /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" CDS complement(223..243) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /note="TEV site" /translation="ENLYFQS" CDS complement(268..285) /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS complement(292..666) /codon_start=1 /locus_tag=" " /product="self-cleaving aspartic endopeptidase SO1698 from Shewanella oneidensis" /note="aspartic endopeptidase" /protein_id=" " /translation="MFAPQGLAQFIKVNVTLENGEPVFIYTDANGQVCQGDITVTQAGT ITYLLNDQTLKGLKFVGVGFVTPFDGIIDAVTISSDGMLVQLVDLDKTPGTTKFQFVLS NTANTLLVLSPDPQIINRPQN" RBS 675..680 /note="ribosome binding site" protein_bind 711..735 /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(736..754) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" promoter 1067..1144 /gene="lacI" /note="lacI promoter" /note=" " CDS 1145..2227 /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 3036..3227 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" rep_origin complement(3657..4245) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4416..5276) /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(5277..5380) /gene="bla" /note="AmpR promoter"
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