Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Tiffany Souterre
Date created: 2012-09-19 11:00:00
Date modified: 2015-05-08 01:13:28
Biobricked pCS2+ plasmid with Elastase promoter for frogs and chicken
| Types | DnaRegion |
| Roles | plasmid_vector Plasmid_Backbone |
| Sequences | BBa_K812200_sequence (Version 1) |
Description
This plasmid is a biobricked version of the plasmid pSC2+(K812000)with Elastase promoter. This plasmid is made of a bacterial ampicilin resistant backbone. The Biobrick site is surrounded by a 5'UTR expression enhancer, and a poly-A tail with a SV40 3' UTR that enhance the stabilisation of the mRNA and the expression of the protein.This plasmid is meant to be micro-injected in frog eggs or chicken eggs. 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
CMV of pCS2+ (K812000) was replaced by Elastase using the double digestion SalI and HindIII. Then J04450 was replaced by sfGFP using the Biobrick K812031 with the double digestion EcoRI and PstI.Source
The plasmid template comes from K812000 and the constitutive CMV promoter was replaced by the Elastase promoter. The sfGFP comes from K812031. sfGFP comes from : Love, N.R. et al., 2011. pTransgenesis: a cross-species, modular transgenesis resource. Development (Cambridge, England), 138(24), pp.5451???8| Sequence Annotation | Location | Component / Role(s) |
| BioBrick suffix T7 prom SV40 Polyadenylation site Downstream polylinker (from pVZ1) T3_promoter M13 primer ColE1 rep origin Ampicillin resistance Bla gene AmpR promoter E.coli f1replication origin Elastase Sp6 primer BioBrick prefix | 1,20 26,42 47,242 243,275 293,312 329,347 679,1346 1494,2344 2396,2424 2503,2809 3012,3224 3232,3249 3292,3312 | feature/BioBrick engineered_region feature/primer_binding primer_binding_site operator feature/operator feature/binding non_covalent_binding_site primer_binding_site feature/primer_binding primer_binding_site feature/primer_binding feature/misc sequence_feature feature/cds CDS feature/promoter promoter sequence_feature feature/misc promoter feature/promoter feature/primer_binding primer_binding_site engineered_region feature/BioBrick |