pPha-DUAL-[2xNR] vector (V004112)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V004112	pPha-DUAL-[2xNR]	In stock, 1 week for quality controls	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pPha-DUAL-[2xNR]

Antibiotic Resistance:: Ampicillin

Length:: 4635 bp

Type:: Shuttle vector

Replication origin:: ori

Source/Author:: Zauner S, Hempel F.

pPha-DUAL-[2xNR] 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.

pPha-DUAL-[2xNR] vector Sequence

LOCUS       40924_34436        4635 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pPha-DUAL-[2xNR], complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4635)
  AUTHORS   Zauner S, Hempel F.
  TITLE     Inducible shuttle vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4635)
  AUTHORS   Zauner S, Hempel F.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JUN-2011) Cell Biology, Philipps-University Marburg, 
            Karl-von-Frisch Str. 8, Marburg 35032, Germany
REFERENCE   3  (bases 1 to 4635)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4635)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-JUN-2011) Cell Biology, Philipps-University Marburg, 
            Karl-von-Frisch Str. 8, Marburg 35032, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      36..451
                     /note="nitrate reductase; derived from Phaeodactylum 
                     tricornutum"
                     /regulatory_class="promoter"
     misc_feature    452..472
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      473..737
                     /note="nitrate reductase; derived from Phaeodactylum 
                     tricornutum"
                     /regulatory_class="terminator"
     regulatory      782..1197
                     /note="nitrate reductase; derived from Phaeodactylum 
                     tricornutum"
                     /regulatory_class="promoter"
     misc_feature    1216..1272
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      1273..1537
                     /note="nitrate reductase; derived from Phaeodactylum 
                     tricornutum"
                     /regulatory_class="terminator"
     regulatory      1546..1731
                     /note="fcpB; derived from Phaeodactylum tricornutum"
                     /regulatory_class="promoter"
     CDS             1732..2103
                     /label=BleoR
                     /note="antibiotic-binding protein"
     regulatory      2107..2348
                     /label=fcpA
                     /note="fcpA"
                     /regulatory_class="terminator"
     promoter        complement(2358..2376)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2634..3222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3396..4253)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4254..4358)
                     /label=AmpR promoter