Basic Vector Information
- Vector Name:
- pKLD116
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6626 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Rocco CJ, Dennison KL, Klenchin VA, Rayment I, Escalante-Semerena JC.
pKLD116 vector Vector Map
pKLD116 vector Sequence
LOCUS 40924_26859 6626 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector pKLD116, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6626) AUTHORS Rocco CJ, Dennison KL, Klenchin VA, Rayment I, Escalante-Semerena JC. TITLE Construction and use of new cloning vectors for the rapid isolation of recombinant proteins from Escherichia coli JOURNAL Plasmid 59 (3), 231-237 (2008) PUBMED 18295882 REFERENCE 2 (bases 1 to 6626) AUTHORS Rocco CJ, Dennison KL, Klenchin VA, Rayment I, Escalante-Semerena JC. TITLE Direct Submission JOURNAL Submitted (10-DEC-2007) Bacteriology, University of Wisconsin - Madison, 1550 Linden Dr., Madison, WI 53706, USA REFERENCE 3 (bases 1 to 6626) TITLE Direct Submission REFERENCE 4 (bases 1 to 6626) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; date: "2008"; volume: "59"; issue: "3"; pages: "231-237" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (10-DEC-2007) Bacteriology, University of Wisconsin - Madison, 1550 Linden Dr., Madison, WI 53706, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6626 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 13..30 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" CDS 37..1134 /codon_start=1 /label=MBP /note="maltose binding protein from E. coli" /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW YAVRTAVINAASGRQTVDEALKDAQT" CDS 1180..1200 /codon_start=1 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" /translation="ENLYFQG" CDS 1269..1286 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" terminator 1353..1400 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1437..1892 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 1919..2023 /label=AmpR promoter CDS 2024..2881 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3055..3643 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(3829..3971) /label=bom /note="basis of mobility region from pBR322" CDS complement(4076..4264) /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" protein_bind complement(5039..5060) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(5076..6155) /codon_start=1 /label=lacI /note="lac repressor" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(6156..6233) /label=lacI promoter promoter 6542..6560 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 6561..6585 /label=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)." RBS 6600..6622 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
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