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pML2424 vector (V017139)

Basic Vector Information

Vector Name:
pML2424
Length:
9513 bp
Type:
Gene knockout
Replication origin:
ori
Host:
Mycobacterium
Promoter:
sacB
Growth Strain(s):
DH5a
Growth Temperature:
30℃

pML2424 vector Vector Map

pML24249513 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200sacB promoterSacBorihygmgfp5loxPdTomato

Plasmid Resuspension 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 ℃.

pML2424 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       62056_17130        9513 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9513)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        194..638
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             639..2057
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     rep_origin      complement(2227..2815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6198..7193)
                     /gene="hyg"
                     /label=hyg
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces 
                     hygroscopicus. Accession#: P09979"
     CDS             complement(7308..8021)
                     /label=mgfp5
                     /note="GFP with folding enhancement mutations"
     protein_bind    complement(8378..8411)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             8796..9497
                     /label=dTomato
                     /note="dimeric variant of DsRed fluorescent protein (Shaner
                     et al., 2004)"

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