Basic Vector Information
- Vector Name:
- pdcDNA-FLAG
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7034 bp
- Type:
- Mammalian vector
- Replication origin:
- ori
- Source/Author:
- De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
- Promoter:
- lac UV5
pdcDNA-FLAG vector Map
pdcDNA-FLAG vector Sequence
LOCUS 40924_14260 7034 bp DNA circular SYN 18-DEC-2018
DEFINITION Mammalian vector pdcDNA-FLAG, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7034)
AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
Fonseca M, Vanhoucke M, Beyaert R.
TITLE BCCM/LMBP Plasmid collection
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7034)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 3 (bases 1 to 7034)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7034)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
9052, BELGIUM"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7034
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 27..406
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 407..610
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 655..673
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 705..719
/codon_start=1
/label=enterokinase site
/note="enterokinase recognition and cleavage site"
/translation="DDDDK"
protein_bind 729..853
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 878..908
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 962..1618
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1963..2265
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2309..2433)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(2451..2469)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 2495..2719
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2765..3193
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3246..3456
/label=SV40 promoter
/note="SV40 early promoter"
CDS 3531..4322
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 4499..4632
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4669..4685)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4693..4709)
/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)."
promoter complement(4717..4747)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4762..4783)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5071..5659)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5833..6690)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6691..6795)
/label=AmpR promoter
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