载体基本信息 | |||
载体名称 | pET-43.1b(+) | 载体抗性 | Ampicillin |
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载体长度 | 7274 bp | 载体类型 | pET & Duet Vectors (Novagen) |
载体来源 | Novagen (EMD Millipore) | 拷贝数 | High copy number |
pET-43.1b(+) 载体载体图谱
pET-43.1b(+) 载体载体序列
LOCUS Exported 7274 bp ds-DNA circular SYN 03-12-2013 DEFINITION Bacterial vector for expressing NusA fusion proteins. For other reading frames, use pET-43.1a(+) or pET-43.1c(+). ACCESSION . VERSION . KEYWORDS pET-43.1b(+) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7274) AUTHORS Novagen (EMD Millipore) TITLE Direct Submission FEATURES Location/Qualifiers source 1..7274 /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" terminator 260..346 /gene="Escherichia coli rrnB" /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(512..529) /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" misc_feature 530..680 /note="MCS" /note="multiple cloning site" CDS complement(536..568) /codon_start=1 /product="HSV (herpes simplex virus) epitope tag" /note="HSV tag" /translation="QPELAPEDPED" CDS complement(680..694) /codon_start=1 /product="enterokinase recognition and cleavage site" /note="enterokinase site" /translation="DDDDK" CDS complement(713..730) /codon_start=1 /product="thrombin recognition and cleavage site" /note="thrombin site" /translation="LVPRGS" CDS complement(746..790) /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /note="S-Tag" /translation="KETAAAKFERQHMDS" CDS complement(800..817) /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS complement(833..2317) /codon_start=1 /gene="nusA" /product="transcription elongation protein (N utilization substance protein A)" /note="NusA" /note="highly soluble in E. coli" /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA" RBS 2326..2331 /note="ribosome binding site" protein_bind 2362..2386 /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(2387..2405) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" promoter 2718..2795 /gene="lacI" /note="lacI promoter" /note=" " CDS 2796..3878 /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 4450..4641 /codon_start=1 /gene="rop" /product="Rop protein" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" rep_origin complement(5071..5659) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS 5833..6693 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin complement(6808..7263) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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