Library of synthetic transcriptional AND gates built with split T7 RNA polymerase mutants.

Shis DL, Bennett MR
Proc Natl Acad Sci U S A. 2013 Mar 26;110(13):5028-33. doi: 10.1073/pnas.1220157110. Epub 2013 Mar 11. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
60717	pTara:500	Expresses T7 RNAP from the arabinose inducible promoter PBAD with a RBS strength of 500 from the Salis RBS calculator. Also includes a constitutive araC ORF
60720	pTSara	Expresses both fragments of T7 RNAP split at position 179. Both fragments are driven by the arabinose inducible promoter PBAD. Also contains a constitutive araC ORF
60721	pTSara-N	Expresses the N-terminal fragment of split T7 RNAP split a position 179. Insert is driven by PBAD. Contains a constitutive araC ORF
60722	pTSara-C	Expresses the C-terminal fragment of split T7 RNAP split a position 179. Insert is driven by the arabinose inducible promoter PBAD. Contains a constitutive araC ORF
60724	pTSara-Q758C	Expresses both fragments of T7 RNAP split at position 179. Both fragments are driven by PBAD. The C-terminal fragment has the point mutation Q758C. Contains a constitutive araC ORF
60725	pTSara-R756S	Expresses both fragments of T7 RNAP split at position 179. Both fragments are driven by PBAD. The C-terminal fragment has the point mutation R756S. Contains a constitutive araC ORF
60726	pTSara-N748D	Expresses both fragments of T7 RNAP split at position 179. Both fragments are driven by PBAD. The C-terminal fragment has the point mutation N748D. Contains a constitutive araC ORF
60727	pTSara-R756K	Expresses both fragments of T7 RNAP split at position 179. Both fragments are driven by PBAD. The C-terminal fragment has the point mutation R756K. Contains a constitutive araC ORF
60728	pTSlb-wt	Expresses both fragments of T7 RNAP split at position 179. The N-terminal fragment is driven by PLac while the C-terminal fragment is driven by PBAD.
60729	pTSlb-Q758C	Expresses both fragments of T7 RNAP split at position 179. The N-terminal fragment is driven by PLac while the C-terminal fragment is driven by PBAD. C-terminal fragment has the point mutations Q758C
60730	pTSlb-R756S	Expresses both fragments of T7 RNAP split at position 179. The N-terminal fragment is driven by PLac while the C-terminal fragment is driven by PBAD. C-terminal fragment has the point mutations R756S
60731	pTSlb-R756K	Expresses both fragments of T7 RNAP split at position 179. The N-terminal fragment is driven by PLac while the C-terminal fragment is driven by PBAD. C-terminal fragment has the point mutations R756S
60732	pTSlb-N748D	Expresses both fragments of T7 RNAP split at position 179. The N-terminal fragment is driven by PLac while the C-terminal fragment is driven by PBAD. C-terminal fragment has the point mutations N748D.
60733	pET28:GFP	pET28 driving expression of GFP, biobrick part Bba_E0040
60734	pET:GACT:GFP	A modified pET28 driving expression of GFP, biobrick part Bba_E0040. No LacO site downstream of PT7. T7 promoter has base pairs 'GACT' between -11 and -8.
60735	pET:GACG:GFP	A modified pET28 driving expression of GFP, biobrick part Bba_E0040. No LacO site downstream of PT7. T7 promoter has base pairs 'GACG' between -11 and -8.
60736	pET:GCAT:GFP	A modified pET28 driving expression of GFP, biobrick part Bba_E0040. No LacO site downstream of PT7. T7 promoter has base pairs 'GCAT' between -11 and -8.
60737	pET:CACT:GFP	A modified pET28 driving expression of GFP, biobrick part Bba_E0040. No LacO site downstream of PT7. T7 promoter has base pairs 'CACT' between -11 and -8.
60738	pET:CCCT:GFP	A modified pET28 driving expression of GFP, biobrick part Bba_E0040. No LacO site downstream of PT7. T7 promoter has base pairs 'CCCT' between -11 and -8.
60740	pET:ACAT:GFP	A modified pET28 driving expression of GFP, biobrick part Bba_E0040. No LacO site downstream of PT7. T7 promoter has base pairs 'ACAT' between -11 and -8.
61151	DH10B-ALT	DH10B modified to constitutively express araC, lacI, and tetR, integrated at the attB site on the E Coli genome. This is DH10B-ALT-Tet with the tetracycline resistance cassette deleted.