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pGM1190 vector (V013330)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013330 pGM1190 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:
pGM1190
Antibiotic Resistance:
Apramycin
Length:
7089 bp
Replication origin:
ori
Host:
Broad host
Selection Marker:
Tsr
Promoter:
TipA
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pGM1190 vector Map

Copy Sequence View Map
pGM11907089 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900oriTApmRoriCAP binding sitelac promoterlac operatorM13 revlambda t0 terminator6xHisthrombin siteT7 tag (gene 10 leader)6xHisfd terminator23S rRNA (adenosine(1067)-2'-O)-methyltransferasepSG5 Rep

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.

pGM1190 vector Sequence

LOCUS       V013330                 7089 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013330
VERSION     V013330
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7089)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(467..576)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             892..1692
                     /label="ApmR"
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     rep_origin      1994..2582
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2870..2891
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2906..2936
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2944..2960
                     /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     2968..2984
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     terminator      complement(3163..3197)
                     /label="lambda t0 terminator"
                     /note="minimal transcription terminator from phage lambda
                     (Scholtissek and Grosse, 1987)"
     CDS             3376..3393
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             3403..3420
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             3424..3456
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             3506..3523
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      complement(3688..3736)
                     /label="fd terminator"
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             4079..4885
                     /gene="tsnR"
                     /label="23S rRNA (adenosine(1067)-2'-O)-methyltransferase"
                     /note="23S rRNA (adenosine(1067)-2'-O)-methyltransferase
                     from Streptomyces azureus. Accession#: P18644"
     CDS             5451..6890
                     /label="pSG5 Rep"
                     /note="replication initiator protein from the Streptomyces
                     ghanaensis plasmid pSG5"