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pUK21 vector (V002289) Gene synthesis in pUK21 backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V002289 pUK21 In stock, instant shipping

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

Basic Vector Information

Vector Name:
pUK21
Antibiotic Resistance:
Kanamycin
Length:
3090 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Vieira J, Messing J.
Growth Strain(s):
Top10
Growth Temperature:
37℃

pUK21 vector Map

Copy Sequence View Map
pUK213090 bp6001200180024003000CAP binding sitelac operatorM13 revSK primerT7 promoterM13 fwdKanRori

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.

pUK21 vector Sequence

LOCUS       40924_45708        3090 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUK21, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3090)
  AUTHORS   Vieira J, Messing J.
  TITLE     New pUC-derived cloning vectors with different selectable markers 
            and DNA replication origins
  JOURNAL   Gene 100, 189-194 (1991)
  PUBMED    1905257
REFERENCE   2  (bases 1 to 3090)
  AUTHORS   Vieira J, Messing J.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-JAN-2000) Waksman Institute, Rutgers University, 190 
            Frelinghuysen Road, Piscataway, NJ 08854, USA
REFERENCE   3  (bases 1 to 3090)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3090)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene 100, 
            189-194 (1991)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-JAN-2000) Waksman Institute, Rutgers University, 190 
            Frelinghuysen Road, Piscataway, NJ 08854, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     protein_bind    183..199
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     207..223
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     351..367
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(382..400)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(407..423)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1375..2175
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="RETSCSTRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDA
                     PELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQ
                     VLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFD
                     DERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRY
                     QDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2321..2909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"