Reporter

Part:BBa_J04450

Designed by: Tamar Odle   Group: iGEM2005   (2005-06-09)


RFP Coding Device

LacI
R0010

B0034
mRFP1
E1010

B0015

The colonies are clearly red in color under natural light after about 18 hours. Smaller colonies are visibly red under UV. The RFP part does not contain a degradation tag and the RBS is strong.

  • LacI sensitive
  • CAP sensitive

This part is commonly used, but can fail if the system contains LacI or CAP protein.
(--Meagan 15:39, 23 July 2009 (UTC))

Team TU_Munich 2012 improved this part by making it compatible to RFC10 and RFC25 (see: BBa_K801100)
(--VolkerMorath 15:02, 21 October 2012 (UTC))

Team NRP-UEA 2013 improved this part by adding a NdeI restriction site before the RFP gene. (see: BBa_K1041000)
(--holusac 20:46, 14 August 2013 (UTC))

Pictures


Usage as a cloning tool

Team Groningen 2010 reports the usage of this part as a cloning tool. When ligating any part, or part assembly, into any standard backbone that contains this part, the non-restricted and single-restricted backbones that self-circularize will produce red colonies on rich media plates (we use TY). These undesired transformants can than be avoided in the screening for the correct construct. With this method, the backbone desired for a new construct does not need to be purified from agarose gel to decrease the amount of undesired tranformants caused by ligation of the original part present in the backbone. The amount of incorrect transformants depends, of course, on the ratio of backbone (mixed with J04450) vs. BioBrick insert, the size of the BioBrick insert, and whether the insert is an assembly of two BioBricks. The images below show two ligations with different efficiencies.

An inefficient assembly ligation of two BioBricks into the pSB1C3 backbone producing many red colonies.
A more efficient, single BioBrick ligation into the pSB1C3 backbone.
[edit]
Categories
//cds/reporter/chromoprotein
//classic/reporter/pret
//function/reporter/color
//function/reporter/pigment
Parameters
emissionRFP
excitation
tagNone