Basic Vector Information
phPK14H 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 ℃.
phPK14H vector Sequence
LOCUS 40924_24822 6815 bp DNA circular SYN 18-DEC-2018
DEFINITION Keratinocyte expression vector phPK14H, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6815)
AUTHORS Hosseini SJ, Zomorodipour A, Jalal R, Sabouni F.
TITLE A recombinant plasmid for specific expression of recombinant protein
in human kertinocytes
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6815)
AUTHORS Hosseini SJ, Zomorodipour A, Jalal R, Sabouni F.
TITLE Keratinocyte specific-expression vector
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 6815)
AUTHORS Hosseini SJ, Zomorodipour A, Jalal R, Sabouni F.
TITLE Direct Submission
JOURNAL Submitted (09-AUG-2006) Molecular Genetics, The National Institute
for Genetic Engineering and Biotechnology, Blvd. Pjoohesh. km-17
Tehran-karaj Highway, Tehran, Tehran 14155-6343, Iran
REFERENCE 4 (bases 1 to 6815)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6815)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(09-AUG-2006) Molecular Genetics, The National Institute for Genetic
Engineering and Biotechnology, Blvd. Pjoohesh. km-17 Tehran-karaj
Highway, Tehran, Tehran 14155-6343, Iran"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6815
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(260..277)
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
misc_feature 2258..2363
/label=multiple cloning site (MCS)
/note="multiple cloning site (MCS)"
promoter complement(2367..2385)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 2411..2635
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2681..3109
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3123..3453
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3520..4311
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 4488..4621
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4658..4674)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4682..4698)
/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(4706..4736)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4751..4772)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5060..5648)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5822..6679)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6680..6784)
/label=AmpR promoter
promoter 6815
/label=K14
/note="Human keratin 14 promoter"
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