载体基本信息 | |||
载体名称 | pET-28a(+) | 载体抗性 | Kanamycin |
---|---|---|---|
载体长度 | 5369 bp | 载体类型 | pET & Duet Vectors (Novagen) |
载体来源 | Novagen (EMD Millipore) | 拷贝数 | High copy number |
pET-28a(+) 载体载体图谱
pET-28a(+) 载体载体序列
LOCUS Exported 5369 bp ds-DNA circular SYN 14-OCT-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Untitled SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5369) TITLE Direct Submission FEATURES Location/Qualifiers source 1..5369 /organism="synthetic DNA construct" /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(207..239) /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /note="T7 tag (gene 10 leader)" /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS complement(243..260) /codon_start=1 /product="thrombin recognition and cleavage site" /note="thrombin site" /translation="LVPRGS" CDS complement(270..287) /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" RBS 306..328 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 343..367 /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(368..386) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" promoter 695..772 /gene="lacI" /note="lacI promoter" CDS 773..1855 /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 2664..2855 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" misc_feature 2957..3099 /note="bom" /note="basis of mobility region from pBR322" rep_origin complement(3285..3873) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS 3995..4810 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /note="KanR" /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin complement(4903..5358) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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