Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: William Rostain, Tristan Cerisy
Date created: 2012-09-17 11:00:00
Date modified: 2015-05-08 01:13:27
Biobricked pCS2+ plasmid for frogs and chicken
| Types | DnaRegion |
| Roles | Plasmid_Backbone plasmid_vector |
| Sequences | BBa_K812000_sequence (Version 1) |
Description
This plasmid is a biobricked version of the standard plasmid pSC2+. This plasmid is made of a bacterial ampicilin reststant backbone, with a universal CMV promoter. The Biobrick site is surrounded by a a 5'UTR expression enhancer, and a ploy-A tail with a SV40 3' UTR that tnhance the stabilisation of the mRNA and the expression of the protein.This plasmid is ment to be micro-injected in frog eggs or chicken eggs for a transcient expression of the protein at the early ages of development of the embryo. This plasmid does not replicate, so it gets diluted and degraded as the animal grow, leaving no chance for DNA propagation to the breeding.
To use this plasmid, you simply have to insert your protein between the EcoRI and PstI site. It is better to add a kozac sequence in front of your gene to enhance its expression.
This plasmid contains the standard registry insert J04450 as a negative cloning control.
Notes
The biobrick site has been inserted in order to obtimize the expression of the protein.Source
This part is the biobricked version of a standard plasmid (see http://sitemaker.umich.edu/dlturner.vectors/home). We obtained this version from Nicolas Pollet lab and it has been biobricked by our team.| Sequence Annotation | Location | Component / Role(s) |
| Biobrick suffix T7 prom (rev) SV40 Polyadenylation site Downstream polylinker (from pVZ1) T3_promoter M13 primer ColE1 rep origin Ampicillin resistance Bla gene AmpR promoter E. coli f1replication origin CMV eukaryotic promoter CMV_fwd_primer Sp6 primer Biobrick prefix | 1,20 26,42 47,242 243,275 293,312 329,347 679,1346 1494,2344 2396,2424 2503,2809 3012,3998 3916,3957 4033,4051 4093,4113 | feature/BioBrick engineered_region feature/primer_binding primer_binding_site feature/operator operator non_covalent_binding_site feature/binding primer_binding_site feature/primer_binding primer_binding_site feature/primer_binding feature/misc sequence_feature feature/cds CDS feature/promoter promoter feature/misc sequence_feature promoter feature/promoter feature/primer_binding primer_binding_site feature/primer_binding primer_binding_site feature/BioBrick engineered_region |