pAP1-Luc vector (V009489)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V009489	pAP1-Luc	In stock, instant shipping	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

The pAP1-Luc is a reporter plasmid for detection of the activator protein 1 transduction pathway.

Vector Name:: pAP1-Luc

Antibiotic Resistance:: Ampicillin

Length:: 5675 bp

Type:: Reporter vector

Replication origin:: ori

Source/Author:: Zheng C-F.

Promoter:: minP

Growth Strain(s):: DH5a

Growth Temperature:: 37℃

pAP1-Luc vector Map

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Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

References

Woodgate A, Walton M, MacGibbon GA, Dragunow M. Inducible transcription factor expression in a cell culture model of apoptosis. Brain Res Mol Brain Res. 1999 Mar 20;66(1-2):211-6.

pAP1-Luc vector Sequence

LOCUS       40924_4574        5675 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pAP1-Luc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  AUTHORS   Zheng C-F.
  TITLE     pAP1-Luc reporter plasmid for detection of the activator protein 1 
            transduction pathway
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5675)
  AUTHORS   Zheng C-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAR-1998) Technical Services, Stratagene, 11011 N. 
            Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 5675)
  AUTHORS   Zheng C-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-OCT-1998) Technical Services, Stratagene, 11011 N. 
            Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE   4  (bases 1 to 5675)
  AUTHORS   Zheng C-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-JAN-1999) Technical Services, Stratagene, 11011 N. 
            Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE   5  (bases 1 to 5675)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 5675)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-MAR-1998) Technical Services, Stratagene, 11011 N. Torrey Pines 
            Rd., La Jolla, CA 92037, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (29-OCT-1998) Technical Services, Stratagene, 11011 N. Torrey Pines 
            Rd., La Jolla, CA 92037, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "Submitted 
            (26-JAN-1999) Technical Services, Stratagene, 11011 N. Torrey Pines 
            Rd., La Jolla, CA 92037, USA"
COMMENT     SGRef: number: 5; type: "Journal Article"
COMMENT     On Jan 26, 1999 this sequence version replaced gi:3808230.
FEATURES             Location/Qualifiers
     source          1..5675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..136
                     /label=AmpR promoter
     CDS             137..994
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1168..1755
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1934..2074)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     polyA_signal    2313..2445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2539..2671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2737..2768
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             2795..4444
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     intron          4578..4643
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    5213..5347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"