pMG36e vector (V012803) Gene synthesis in pMG36e backbone

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Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V012803	pMG36e	In stock, instant shipping

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

Basic Vector Information

pMG36e plasmid (pWV01 ori) is based on protease gene transcription and translation signals from Lactococcus lactis cremoris, is a typical artificial constitutive expression vector as a main expression system in Lactococcus.It contains a strong promoter and can express foreign proteins in a variety of bacteria. It has been used to study the action mechanism of bacteriocin, the transformation of lactic acid bacteria genetic engineering strains and the development of oral vaccines. It has been widely used in many fields and has become one of the important tools for lactic acid bacteria genetic engineering research.

Please note:
1.The competent cell should be MC1061. MC1061/P3 Chemically Competent E. coli are used for highly efficient transformations that require the P3 episome for selection and maintenance of vectors encoding the tyrosine tRNA suppressor (synthetic supF gene, such as pCDM8, pcDNA1.1, or any other supF-containing vector). TOP10 is not a MC1061 derivative;
2. The Erythromycin selection should be 200 ug/mL. Erythromycin is very tricky, and 40-50ug/mL will not suppress non-target bacteria.
3. The culture temperature should be 30℃.

Vector Name:: pMG36e

Antibiotic Resistance:: Erythromycin

Length:: 3600 bp

Type:: Lactococcus lactis expression plasmid

Replication origin:: pWV01

Copy Number:: High copy number

Promoter:: P32

Cloning Method:: Enzyme Cut

Growth Strain(s):: MC1061

Growth Temperature:: 30℃

pMG36e vector Map

Copy Sequence View Map

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.

References

Yue M, Wei J, Chen W, Hong D, Chen T, Fang X. Neurotrophic Role of the Next-Generation Probiotic Strain L. lactis MG1363-pMG36e-GLP-1 on Parkinson's Disease via Inhibiting Ferroptosis. Nutrients. 2022;14(22):4886. Published 2022 Nov 18. doi:10.3390/nu14224886

pMG36e vector Sequence

LOCUS       Exported                3600 bp DNA     circular SYN 03-DEC-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pMG36e
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3600)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3600)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3600)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3600)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             221..919
                     /codon_start=1
                     /label=repA
                     /note="repA"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDCYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     CDS             complement(1454..2188)
                     /codon_start=1
                     /product="23S rRNA (adenine(2058)-N(6))-methyltransferase, 
                     also known as Erm methyltransferase, is an enzyme that 
                     confers resistance to erythromycin and other macrolide, 
                     lincosamide, and streptogramin B (MLSB) antibiotics. It 
                     works by methylating the adenine at position 2058 of the 
                     23S ribosomal RNA, which prevents erythromycin from binding
                     to the ribosome and inhibiting protein synthesis. "
                     /label=Emr
                     /note="Emr"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
                     NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     promoter        2465..2643
                     /note="P32"
     repeat_region   3065..3195
                     /label=IRI repeat region
                     /note="A cluster of identical repeat sequences (IRI) that
                     likely functions in genome organization, structural 
                     maintenance, or regulatory processes through repeated DNA 
                     elements.; id: 4274850"
     CDS             complement(3204..3215)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   3242..3264
                     /label=IRII Inverted Repeat Region
                     /note="A cluster of identical inverted repeat sequences
                     (IRII) that likely function in DNA replication initiation, 
                     recombination, or regulatory processes by forming stem-loop
                     structures.; id: 4274849"
     repeat_region   3348..3396
                     /label=IRIII Repeat Region
                     /note="A cluster of inverted repeat sequences (IRIII) that 
                     likely function in DNA replication initiation, 
                     recombination regulation, or structural organization of 
                     genetic elements.; id: 4274848"
     CDS             3349..3558
                     /codon_start=1
                     /label=repC
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITKR"

.cls { stroke-width: 0px; fill: url(#c6); } pMG36e vector (V012803) Gene synthesis in pMG36e backbone