This part came from two previous parts J119000 and J119009.
This part consists of both T7RNAP and the T7RNAP promoter. The gene produces a blue color when on a plate that contains X-gal. The gene produces T7RNAP which attaches to the T7RNAP promoter this produces LacZ and a blue color.
We used parts I732017 and R0085 to construct the pT7-rbs-lacZ reporter.
When inserted, cells will have a functioning beta-galactosidase gene. If placed onto a plate with X-Gal, cells with this functioning part will split apart the X-gal and colonies will appear blue.
pLacI-RBS-T7RNAP with sites for oligo insertion
J23100 promoter and B0030 RBS upstream of T7 RNAP gene. The following restriction sites do not occur in the promoter-RBS-T7 RNAP sequence. Some of them did not occur in the original sequence and others were removed while conserving the amino acid sequence. Both the forward and reverse complement were checked for type IIs enzyme sites. EcoRI, XbaI, SpeI, PstI, AscI, AsiSI, BsaI, BsmBI, BbsI
BsmBI site (cgtctcn/ and /nnnnngagacg) before RBS and after gene coding sequence that can be used to cut out RBS-gene for placement after other promoters. Sites for AsiSI (gcgat/cgc) and AscI (gg/cgcgcc) that can be used to insert existing oligos. Sites flanking these for BsaI (ggtctcn/ and /nnnnngagacc) that can be used to insert new oligos without the need for gel purification. A site for BbsI (gaagacnn/ and /nnnnnngtcttc) that produces a sticky end complementary to the first BsaI site, for use in generating a near wild type version of the gene.
The T7 promoter should only produce PoPS when the T7 polymerase is also being expressed.
RBS (Elowitz 1999) -- defines RBS efficiency
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
lacZ (encoding beta-galactosidase, full-length)
PCR amplified and standardized from BL21
LacZ encodes beta-galactosidase, an intracellular enzyme that cleaves the disaccharide lactose into glucose and galactose. Beta-galactosidase can also cleaves X-gal and ONPG into colorful products.
Conversion of the iGEM parts registry to SBOL2.1