pnlslacZ-ACN vector (V004298)

Basic Vector Information

Vector Name:
pnlslacZ-ACN
Antibiotic Resistance:
Ampicillin
Length:
9675 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Wu S, Ying G, Wu Q, Capecchi MR.
Promoter:
SP6

pnlslacZ-ACN vector Vector Map

pnlslacZ-ACN9675 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600T7 promoterSV40 NLSbeta-globin poly(A)loxPHSV TK poly(A) signalNeoR/KanRRNA polymerase II large subunit promoterHSV TK poly(A) signalsmall t intronSV40 NLSWELQut siteloxP siteSP6 promoterAmpR promoterAmpRori

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 ℃.

pnlslacZ-ACN vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_33402        9675 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pnlslacZ-ACN, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9675)
  AUTHORS   Wu S, Ying G, Wu Q, Capecchi MR.
  TITLE     A protocol for constructing gene targeting vectors: generating 
            knockout mice for the cadherin family and beyond
  JOURNAL   Nat Protoc 3 (6), 1056-1076 (2008)
  PUBMED    18546598
REFERENCE   2  (bases 1 to 9675)
  AUTHORS   Wu S, Capecchi MR.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-FEB-2008) Howard Hughes Medical Institute, University 
            of Utah, 15N 2030E, Salt Lake City, UT 84112, USA
REFERENCE   3  (bases 1 to 9675)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9675)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Protoc"; date: "2008"; volume: "3"; issue: "6"; pages: "1056-1076"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-FEB-2008) Howard Hughes Medical Institute, University of Utah, 
            15N 2030E, Salt Lake City, UT 84112, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..52
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             315..335
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    3184..3578
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     protein_bind    3590..3623
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3650..3698
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      complement(3650..3697)
                     /label=HSV thymidine kinase polyA signal
                     /note="HSV thymidine kinase polyA signal"
                     /regulatory_class="polyA_signal_sequence"
     CDS             complement(3708..4508)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     regulatory      complement(4525..5231)
                     /label=RNA polymerase II large subunit promoter
                     /note="RNA polymerase II large subunit promoter"
                     /regulatory_class="promoter"
     polyA_signal    complement(5247..5295)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     intron          complement(5645..5710)
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(6536..6556)
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             complement(6580..6591)
                     /label=WELQut site
                     /note="WELQut protease recognition and cleavage site"
     misc_recomb     7266..7299
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(7266..7299)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(7363..7381)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        7727..7831
                     /label=AmpR promoter
     CDS             7832..8689
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8863..9451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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