Positive and negative feedback regulated T7 RNAP expression system
Based on and developed from a system built by Bill Studier at Brookhaven National Labs
This part is an intermediate version of a biological virtual machine. T7 RNAP expression is controlled by a T7 promoter, forming a positive feedback loop or autogene. LacI expression is controlled by the same promoter. The promoter is repressible by LacI leading to negative feedback. The combination of positive and negative feedback should lead to a moderate, stable level of T7 RNAP in the cell.
Double protein generator (T7 RNAP and LacI)
LacI and T7 RNAP are transcribed polycistronically and translation of both is regulated by B0032. A T7 terminator is downstream. This part is being used in the construction of VM2.0
This sequence is based on the T7lac promoter described by Dubendorff and Studier (JMB 1991, 219, 45-59). The only modification is the mutation at -16.
A mutant version of the T7lac promoter designed by Dubendorff and Studier. The promoter is based on the strong ??10 T7 promoter but incorporates an A->C mutation at -16. The LacI operator site is a partially symmetric 25bp sequence.
The -16 mutation was included to weaken the consensus promoter as described by Imburgio and co-workers (see BBa_R0183 for more information)
This protein generator is composed of an E. coli RBS, a lacI coding region and a T7 terminator. It is designed for use with a T7 promoter but should be functional with a E. coli promoter also. Ultimately, it will be used in an early version of a virtual machine.
B0032 is a very reliable RBS and B0016 is the T7 terminator
RBS.3 (medium) -- derivative of BBa_0030
Weak1 RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-2" in figure 4-14 of thesis). Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
LacI repressor protein, without SsrA degradation tag. No Barcode.
Bacteriophage T7
Gene 1 coding region from bacteriophage T7 encoding a RNAP polymerase (RNAP). This single-protein RNAP recognizes a 21 bp consensus promoter sequence and is orthogonal to E. coli RNAP.
T7 gene 1 specific transcriptional terminator
Conversion of the iGEM parts registry to SBOL2.1