Basic Vector Information
FLAG.PKCepsilon vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
FLAG.PKCepsilon vector Sequence
LOCUS Exported 7616 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS FLAG.PKCepsilon
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7616)
AUTHORS Cenni V, Doppler H, Sonnenburg ED, Maraldi N, Newton AC, Toker A
TITLE Regulation of novel protein kinase C epsilon by phosphorylation.
JOURNAL Biochem J. 2002 May 1. 363(Pt 3):537-45.
PUBMED 11964154
REFERENCE 2 (bases 1 to 7616)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biochem J.
2002 May 1. 363(Pt 3):537-45."
FEATURES Location/Qualifiers
source 1..7616
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(42..61)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 233..612
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 613..816
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 767..787
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 861..880
/label=T7
/note="T7 promoter, forward primer"
promoter 861..879
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 917..940
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1585..1596
/codon_start=1
/gene="cspA"
/product="translation enhancing element for E. coli (Qing
et al., 2004)"
/label=TEE
/translation="NHKV"
promoter complement(3167..3185)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3168..3185)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3205..3222)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3211..3435
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3481..3909
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3568..3587)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3778..3799
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3918..3938)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3923..4252
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4103..4238
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4165..4184
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4319..5113
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4373..4392)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4983..5002
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5287..5408
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5324..5343)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5378..5397
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5457..5473)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5457..5473)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5470..5492)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5481..5497
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
promoter complement(5505..5535)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5550..5571
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5688..5705)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5859..6447)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5939..5958)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6618..7478)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7241..7260
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7479..7583)
/gene="bla"
/label=AmpR promoter
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