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pUCDMIG vector (V002382) Gene synthesis in pUCDMIG backbone

Price Information

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V002382 pUCDMIG In stock, instant shipping

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

Basic Vector Information

Vector Name:
pUCDMIG
Antibiotic Resistance:
Chloramphenicol
Length:
4279 bp
Type:
Baculovirus expression vector
Replication origin:
R6K γ ori
Source/Author:
Yao L, Sun J.
Promoter:
p10
Growth Strain(s):
GT115
Growth Temperature:
37℃

pUCDMIG vector Map

Copy Sequence View Map
pUCDMIG4279 bp600120018002400300036004200R6K gamma oriloxPHSV TK poly(A) signalp10 promoterpolyhedrin promoter5'UTREGFPSV40 poly(A) signalT7 terminatorcat promoterCmR

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.

pUCDMIG vector Sequence

LOCUS       40924_45253        4279 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Baculovirus expression vector pUCDMIG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4279)
  AUTHORS   Yao L, Sun J.
  TITLE     Construction of a baculovirus-silkworm multigene expression system 
            and its application on producing virus-like particles
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4279)
  AUTHORS   Yao L, Sun J.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-AUG-2011) Hen Nan Provincial Key Laboratory of Funiu 
            Mountain Insect Biology, Nan Yang Normal University, 1638 Wolong 
            Road, Nan Yang, He Nan 473061, China
REFERENCE   3  (bases 1 to 4279)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4279)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-AUG-2011) Hen Nan Provincial Key Laboratory of Funiu Mountain 
            Insect Biology, Nan Yang Normal University, 1638 Wolong Road, Nan 
            Yang, He Nan 473061, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(6..394)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(415..448)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    complement(638..686)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(822..931)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        976..1067
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     5'UTR           1175..1754
                     /note="5'-UTR internal ribosome entry site (IRES) sequence
                     of Rhopalosiphum padi virus"
     CDS             1764..2480
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    2626..2760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      2846..2893
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3131..3233
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             3234..3890
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"