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pTNT-B18R vector (V000975)

Price Information

Cat No. Plasmid Name Availability Add to cart
V000975 pTNT-B18R In stock, 1 week for quality controls

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

Basic Vector Information

Vector Name:
pTNT-B18R
Antibiotic Resistance:
Ampicillin
Length:
3907 bp
Type:
Bacterial Expression
Replication origin:
ori
Promoter:
SP6
Cloning Method:
Restriction Enzyme
5' Primer:
SP6

pTNT-B18R vector Map

Copy Sequence View Map
pTNT-B18R3907 bp60012001800240030003600T7 promoterSP6 promoterKozak sequenceSoluble interferon alpha/beta receptor OPG204T7 terminatorf1 oripRS-markerpGEX 3'pBRforEcoAmpR promoterAmpRori

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.

pTNT-B18R vector Sequence

LOCUS       V000975                 3907 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000975
VERSION     V000975
KEYWORDS    pTNT-B18R
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 3907)
  AUTHORS   Yoshioka N, Gros E, Li HR, Kumar S, Deacon DC, Maron C, Muotri AR,
            Chi NC, Fu XD, Yu BD, Dowdy SF
  TITLE     Efficient generation of human iPSCs by a synthetic self-replicative
            RNA.
  JOURNAL   Cell Stem Cell. 2013 Aug 1;13(2):246-54. doi:
            10.1016/j.stem.2013.06.001.
   PUBMED   23910086
REFERENCE   2  (bases 1 to 3907)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3907)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi:
            "10.1016/j.stem.2013.06"; journalName: "Cell Stem Cell"; date:
            "2013-08-1- 1"; volume: "13"; issue: "2"; pages: "246-54"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..34
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        40..58
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      94..103
                     /label="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             106..1155
                     /gene="OPG204"
                     /label="Soluble interferon alpha/beta receptor OPG204"
                     /note="Soluble interferon alpha/beta receptor OPG204 from
                     Vaccinia virus (strain Western Reserve). Accession#:
                     P25213"
     terminator      1197..1244
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     rep_origin      1323..1778
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1845..1864)
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     1964..1986
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2024..2042)
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        2110..2214
                     /label="AmpR promoter"
     CDS             2215..3072
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      3246..3834
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"