Plasmid Modules

Antibiotic marker

This module includes the structural gene for the antibiotic of choice with its own native promoter. This segment is flanked by unique SwaI and PshAI restriction sites.

Available antibiotic markers:

  • Ampicillin
  • Kanamycin
  • Chloramphenicol
  • Streptomycin/Spectinomycin
  • Tetracycline
  • Gentamicin

Origin of replication

This module comprises the oriV sequence and the replication proteins, and it is flanked by the FseI and AscI restriction sites.

Available replication origins:

  • R6K
  • RK2
  • pBBR1
  • pRO1600/ColE1
  • RFS1010
  • p15A
  • pSC101
  • pUC
  • pBR322/ROP

Cargo

This DNA portion confers functionality to the vectors. This region is formatted as a DNA segment flanked by PacI and SpeI sites.

Available cargo types:

  • Cloning (MCS, lacZ-alpha and lacZ-alpha-I-SceI)
  • Expression systems (lacIq-Ptrc and xylS-Pm)
  • Promoter-probe (lacZ, luxCDABE, YFP, CFP, DsRed2, and mCherry)

 

 
 

Gadgets

Gadgets are DNA sequences that endow a new functionality or property to the basic frame of the SEVA plasmids. The extant structure of the vectors allow addition of up to 6 new gadgets of the kind, as this is the number of unusual (and unique) restriction sites that punctuate the intervening regions between the 3 primary modules. Such sites are in principle permissive for insertions of such functional sequences, although only one of them (SanDI-SwaI) has been used thus far.