Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Tristan Cerisy
Date created: 2012-09-19 11:00:00
Date modified: 2015-05-08 01:13:28
Biobricked pCS2+ plasmid with HSP70 promoter for frogs and chicken
| Types | DnaRegion |
| Roles | plasmid_vector Plasmid_Backbone |
| Sequences | BBa_K812300_sequence (Version 1) |
Description
This plasmid is a biobricked version of the plasmid pSC2+(K812000)with HSP70 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.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 HSP70 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 is from K812000, The constitutive CMV promoter was replaced by Heat Shock Promoter 70, and sfGFP is from K812031. HSP70 is 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 promoter (rev) SV40 Polyadenylation site downstream polylinker (from pVZ1) T3_promoter M13 primer ColE1 rep origin Ampicillin resistant Bla gene AmpR promoter E. coli f1 replication origin HSP70 SP6 primer prefix | 1,20 26,42 47,242 243,275 293,312 329,347 679,1346 1494,2344 2396,2424 2503,2809 3017,3503 3511,3528 3571,3591 | feature/BioBrick engineered_region feature/primer_binding primer_binding_site operator feature/operator non_covalent_binding_site feature/binding primer_binding_site feature/primer_binding primer_binding_site feature/primer_binding sequence_feature feature/misc CDS feature/cds feature/promoter promoter feature/misc sequence_feature feature/promoter promoter non_covalent_binding_site feature/binding feature/BioBrick engineered_region |