载体基本信息 | |||
载体名称 | pET-42b(+) | 载体抗性 | Kanamycin |
---|---|---|---|
载体长度 | 5929 bp | 载体类型 | pET & Duet Vectors (Novagen) |
载体来源 | Novagen (EMD Millipore) | 拷贝数 | High copy number |
pET-42b(+) 载体载体图谱
pET-42b(+) 载体载体序列
LOCUS Exported 5929 bp ds-DNA circular SYN 02-12-2013 DEFINITION Bacterial vector for expressing GST fusion proteins with a Factor Xa site. For other reading frames, use pET-42a(+) or pET-42c(+). ACCESSION . VERSION . KEYWORDS pET-42b(+) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5929) AUTHORS Novagen (EMD Millipore) TITLE Direct Submission FEATURES Location/Qualifiers source 1..5929 /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(150..173) /codon_start=1 /product="8xHis affinity tag" /note="8xHis" /translation="HHHHHHHH" misc_feature 174..263 /note="MCS" /note="multiple cloning site" CDS complement(264..275) /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" CDS complement(306..350) /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /note="S-Tag" /translation="KETAAAKFERQHMDS" CDS complement(366..383) /codon_start=1 /product="thrombin recognition and cleavage site" /note="thrombin site" /translation="LVPRGS" CDS complement(393..410) /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS complement(438..1091) /codon_start=1 /gene=" " /product="glutathione S-transferase from Schistosoma japonicum" /note="GST" /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" RBS 1100..1105 /note="ribosome binding site" protein_bind 1136..1160 /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(1161..1179) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" promoter 1492..1569 /gene="lacI" /note="lacI promoter" /note=" " CDS 1570..2652 /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 3224..3415 /codon_start=1 /gene="rop" /product="Rop protein" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" rep_origin complement(3845..4433) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS 4555..5370 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /note="KanR" /note="confers resistance to kanamycin" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin complement(5463..5918) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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