Vmax has a low-level natural resistance to kanamycin but is more susceptible to other antibiotics. When selecting on Kan plates, Vmax with KanR will simply grow faster than Vmax without KanR. Larger colonies=KanR. Other antibiotics select against Vmax but must have a dosage change compared to E. coli.

Wash the culture in fresh LB before adding glycerol, to avoid problems with reviving the culture later

ecovery media (5 BHI + v2 + 68 mM Sucrose )

Four inducible promoters were found to function in V. natriegens

Vmax@SGIDNA.com

To prepare V. natriegens cells for −80 °C storage, an overnight culture of V. natriegens was washed in fresh medium before storage in glycerol

For best results, use only LB-Miller agar plates. Do not use LB-Lennox or LB-Luriaplates (Vmax™ growth will be suboptimal).

Vibrio natriegens strain containing a major extracellular nuclease knockout and insertion of an IPTG-inducible T7 RNA polymerase cassette for expression of genes under a T7 promoter.

(a)colE1(b)SC101(c)RSF1010

we successfully demonstrated challenging clonings such as Gibson Assembly with 5 and Aquacloning with 3 fragments as well as 8 part golden-gate reactions.

all downstream manipulations with V. natriegens ATCC14048 Δdns cells were performed with antibiotic selection (kanamycin (200 μg/mL), chloramphenicol (2 μg/mL), carbenicillin (100 μg/mL))

V. natriegens grows rapidly in BHIN complex medium with a μ of up to 4.43 h−1 (doubling time of 9.4 min) as well as in minimal medium supplemented with various industrially relevant substrates.

Antibiotic concentrations used for plasmid selection in V. natriegens: ampicillin–carbenicillin 100 µg ml−1, kanamycin 75 µg ml−1, chloramphenicol 5 µg ml−1, spectinomycin 100 µg ml−1. E. coli experiments were performed in standard LB and M9 media.