Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jara Radeck
Date created: 2012-09-22 11:00:00
Date modified: 2015-05-08 01:13:30
Sporovector
| Types | DnaRegion |
| Roles | Plasmid plasmid |
| Sequences | BBa_K823054_sequence (Version 1) |
Description
For N-terminal fusion proteins with cgeA (display on B. subtilis spores).==Sporovector==Our Sporovector is specially designed to give you the opportunity to easily create Sporobeads with any proteins you can think of. It is available as BioBrick in pSB1C3 and can be cloned in any of our empty B. subtilis vectors. We also offer it ???precloned??? in pSBBs4S which in this version lacks NgoMIV restriction sites.
The Sporovector was created by ligation of three cut PCR-products. It is designed to allow N-terminal fusion of genes in Freiburg standard (RCF25). Therefore, the gene to be inserted must be cut with XbaI and AgeI and the vector must be digested with XbaI and NgoMIV. By this digest, the RFP cassette will be removed from this vector to allow easy selection for the integration of the new insert. After ligation, there is a scar of six nucleotides between both genes but no restriction site left.
The gene fusion is controlled by the PcotYZ-promoter which was the strongest one in our promoter evaluations. The C-terminal gene part is cgeA, one of the two known spore crust proteins.
The vector is cloned but has not been tested with an insert, yet.
We are working on further Sporovectors to offer also fusion with cotZ as well as C-terminal fusions.
Notes
For N-terminal fusion proteins with cgeA (display on B. subtilis spores)Source
Assembly of several PCRs.| Sequence Annotation | Location | Component / Role(s) |
| hom thrC EcoRI PcotYZ XbaI RFP NgoMIV cgeA AgeI B0014 PstI spec thrC thrB bla hom | 1,247 247,756 766,772 773,958 959,964 1192,1872 2035,2040 2038,2439 2431,2436 2448,2542 2543,2548 2738,3412 3555,3983 3980,4390 5243,6103 6258,6313 | feature/cds CDS feature/cds CDS sequence_feature feature/dna feature/promoter promoter sequence_feature feature/dna feature/cds CDS sequence_feature feature/dna CDS feature/cds sequence_feature feature/dna stem_loop feature/stem_loop sequence_feature feature/dna feature/cds CDS feature/cds CDS feature/cds CDS feature/cds CDS CDS feature/cds |