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pOTENT-1 vector (V001455)

Price Information

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V001455 pOTENT-1 In stock, instant shipping

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

Basic Vector Information

Adenoviral pOTENT-1 shuttle vector with kanamycin selection and CMV promoter and C-terminal FLAG and His tags.

Vector Name:
pOTENT-1
Antibiotic Resistance:
Kanamycin
Length:
7509 bp
Type:
Mammalian Expression Vectors
Replication origin:
ori
Selection Marker:
Puromycin
Copy Number:
Low copy number
Promoter:
CMV
Cloning Method:
Enzyme Cut
5' Primer:
T7 promoter
3' Primer:
pOT-R
Fusion Tag:
FLAG/His
Expression Method:
Constiutive, Stable / Transient

pOTENT-1 vector Map

Copy Sequence View Map
pOTENT-17509 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500ITRAd5 PsiCMV enhancerCMV promoterT7 promoterFLAG6xHisSV40 poly(A) signalHexon-interlacing proteinSV40 promoterPuroRbGH poly(A) signalEarly 4 ORF1 proteinITRoriNeoR/KanRM13 ori

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.

pOTENT-1 vector Sequence

LOCUS       V001455                 7509 bp    DNA     circular SYN 08-SEP-2020
DEFINITION  Exported.
ACCESSION   V001455
VERSION     V001455
KEYWORDS    pOTENT-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7509)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7509)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   21..123
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     misc_signal     211..361
                     /label="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        419..722
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        954..972
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1073..1096
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1106..1123
                     /label="6xHis"
                     /note="6xHis affinity tag"
     polyA_signal    1294..1415
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     CDS             1509..1928
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     promoter        2342..2671
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             2729..3325
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     polyA_signal    3329..3553
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(3771..4154)
                     /note="Early 4 ORF1 protein from Human adenovirus C
                     serotype 2. Accession#: P03242"
                     /label="Early 4 ORF1 protein"
     repeat_region   4464..4566
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     rep_origin      complement(4838..5426)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             6258..7049
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(7100..7480)
                     /direction=LEFT
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"