Basic Vector Information
- Vector Name:
- pFN19K HaloTag T7 SP6
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4421 bp
- Type:
- Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- Promega
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- SP6
pFN19K HaloTag T7 SP6 vector Vector Map
pFN19K HaloTag T7 SP6 vector Sequence
LOCUS 40924_20301 4421 bp DNA circular SYN 01-JAN-1980 DEFINITION Flexi(R) vector with a kanamycin resistance marker, for cell-free expression of a protein with a cleavable N-terminal HaloTag(R). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4421) AUTHORS Promega TITLE Direct Submission REFERENCE 2 (bases 1 to 4421) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT Subclone a coding sequence between the SgfI/AsiSI and PmeI sites to remove the lethal barnase gene. FEATURES Location/Qualifiers source 1..4421 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 21..39 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 45..63 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" CDS 80..970 /codon_start=1 /label=HaloTag(R) /note="modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives" /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL IGSEIARWLSTLEISG" CDS 983..1003 /codon_start=1 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" /translation="EDLYFQS" CDS 1041..1373 /codon_start=1 /label=barnase /note="ribonuclease from Bacillus amyloliquefaciens" /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY QTFTKIR" misc_feature 1386..1441 /label=MCS /note="pUC18/19 multiple cloning site" terminator 1544..1591 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS 1972..2763 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase from Tn5" /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" rep_origin complement(2938..3526) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 3642..3925 /label=cer region /note="ColE1-derived recombination site that helps to maintain plasmids as monomers" terminator 4155..4241 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4333..4360 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene"
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