Part:BBa_K2033011
N-Acyl Homoserine Lactone (AHL) Modular Sender Vector
This is a modular sender vector separate from psB1C3 designed as a standard backbone for most AHL Senders. The part begins with a pTet promoter, followed by 2 ribosome binding sites, followed by mCherry and a terminator. The AHL synthase will insert between the two RBSs. Ampicillin Resistance (AmpR) is also on the plasmid.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Brief Background
N-acyl homoserine lactones (AHLs) are a category of chemical communication used by bacteria to conduct quorum sensing. AHL quorum sensing results under certain environmental conditions (usually cellular density), causing the AHL synthase to produce a specialized AHL molecule that is recognized by a transcription factor. The shared structure of AHLs consists of a lactone ring and an acyl tail, as shown below:
Synthetic AHL quorum sensing functions within two modules. The first module, the "Sender," must be induced by certain environmental conditions, usually population density of surrounding organisms. This will begin production of the AHL by an AHL synthase, which is then detected by the second module, the "Receiver." The Receiver will cause the expression or silencing of certain genes to achieve the desired purpose of the communication, whether it is the production of GFP or to increase growth rate.
Modular Sender Vector
Our team designed this plasmid backbone as a cassette for AHL Sender genes. This part is a non-standard vector that is to be used in place of psB1C3. The composite part shown on this page is not the complete plasmid-it simply depicts the portion of vector composed of registered parts. The backbone contains a constitutive Tet promoter, two B0034 ribosome binding sites, mCherry, and a B0015 terminator. An RFC BioBrick prefix is added between the two RBSs as the site of insertion for the AHL synthase, while ampicillin resistance is also located on the plasmid. The total plasmid size will be about 3800bp. A plasmid map is shown below:
A more detailed plasmid map can be seen below in this Benchling Link. We also submitted this to the DNASU plasmid repository, which can be found here DNASU.
This Modular Sender Vector, or MSV, permits the user to clone AHL Senders into a standard backbone which provides a visual indicator (mCherry), antibiotic resistance(ampR) and effective RBSs. The RFC prefix may be cut by EcoRI and XbaI in order to insert the AHL part, which should be digested with EcoRI and SpeI. Any potential inserts should not have EcoRI, XbaI, SpeI, or NotI cut sites. This vector has been shown to be compatible with a BL21 or DH5αT E. coli chassis, and is capable of constitutive AHL production for 10 AHL systems, as shown [http://2016.igem.org/Team:Arizona_State/Results here].
| None |