Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Tom Knight
Date created: 2009-05-05 11:00:00
Date modified: 2015-05-08 01:08:20
constitutive cat cassette for Mesoplasma florum
| Types | DnaRegion |
| Roles | engineered_region Generator |
| Sequences | BBa_J70304_sequence (Version 1) |
Description
This part is a version of J70026, the cat cassette from pC194. The original cassette is inducible, with a 5' UTR containing a hairpin structure which sequesters the Shine-Dalgarno sequence. In this mutation, we eliminate the hairpin structure from the 5' UTR to attempt a constitutive construct.Notes
Test for KpnI sites in the source plasmid.Source
PCR from pSB1T3-J70026 with primersGTTTCTTCggtacctttcttcattagaatcaatcc,cat-mut-R
GTTTCTTCggtacctttaggaggcatatcaaatg,cat-mut-F
These primers bind to the source plasmid and remove the left side of the hairpin and add a pair of KpnI sites, which will be cut and religated. I hope to select on Cm plates, since the construct should be Cm resistant.
| Sequence Annotation | Location | Component / Role(s) |
| UP sequence -35 -10 rbs cat terminator KpnI site | 157,179 192,197 215,220 237,241 251,901 914,936 227,233 | feature/misc sequence_feature feature/promoter promoter feature/promoter promoter ribosome_entry_site feature/rbs feature/cds CDS feature/stem_loop stem_loop sequence_feature feature/misc |