Basic Vector Information
- Vector Name:
- pNT1.1 for cKO
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7104 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Tokuhiro K, Ikawa M, Benham AM, Okabe M.
- Promoter:
- mPGK
pNT1.1 for cKO 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 ℃.
pNT1.1 for cKO vector Sequence
LOCUS 40924_33542 7104 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pNT1.1 for cKO, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7104)
AUTHORS Tokuhiro K, Ikawa M, Benham AM, Okabe M.
TITLE Protein disulfide isomerase homolog PDILT is required for quality
control of sperm membrane protein ADAM3 and male fertility
[corrected]
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 109 (10), 3850-3855 (2012)
PUBMED 22357757
REFERENCE 2 (bases 1 to 7104)
AUTHORS Ikawa M.
TITLE Direct Submission
JOURNAL Submitted (26-OCT-2011) Rimd, Osaka University, 3-1 Yamada-Oka,
Suita, Osaka 5650871, Japan
REFERENCE 3 (bases 1 to 7104)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7104)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
Acad. Sci. U.S.A."; date: "2012"; volume: "109"; issue: "10"; pages:
"3850-3855"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-OCT-2011) Rimd, Osaka University, 3-1 Yamada-Oka, Suita, Osaka
5650871, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7104
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 7..61
/note="multi-cloning site 1 (PmeI, NotI, SalI, NsiI, AsiSI,
XhoI, SfiI)"
misc_feature 91..124
/label=LoxP site 1
/note="LoxP site 1"
protein_bind 91..124
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 137..142
/label=BamHI
/note="BamHI"
misc_feature 146..193
/label=FRT site 1
/note="FRT site 1"
protein_bind complement(146..193)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
polyA_signal complement(203..251)
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(261..1061)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
promoter complement(1075..1574)
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
misc_feature 1601..1606
/label=HindIII
/note="HindIII"
protein_bind complement(1610..1657)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_feature 1661..1686
/label=multi-cloning site 2 (EcoRI, NheI, AflII, AscI)
/note="multi-cloning site 2 (EcoRI, NheI, AflII, AscI)"
protein_bind complement(1687..1720)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 1721..1750
/label=multi-cloning site 3 (PacI, KpnI, MfeI, AvrII)
/note="multi-cloning site 3 (PacI, KpnI, MfeI, AvrII)"
promoter 1765..2264
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
5'UTR 2306..2345
/label=HSV TK 5'-UTR
/note="5' untranslated region from the herpes simplex virus
thymidine kinase gene"
CDS 2363..3490
/label=HSV TK
/note="herpes simplex virus thymidine kinase"
polyA_signal 3535..3583
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
polyA_signal 4100..4554
/label=PGK poly(A) signal
/note="mouse phosphoglycerate kinase 1 polyadenylation
signal"
misc_feature 4558..4563
/label=ClaI
/note="ClaI"
promoter 4927..5031
/label=AmpR promoter
CDS 5032..5889
/label=AmpR
/note="beta-lactamase"
rep_origin 6063..6651
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 6939..6960
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6975..7005
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7013..7029
/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)."
primer_bind 7037..7053
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 7074..7092
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
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