BBa_K1857002

BBa_K1857002 Version 1

Component

Source: http://parts.igem.org/Part:BBa_K1857002

Generated By: https://synbiohub.org/public/igem/igem2sbol/1

Created by: Joshua Timmons

Date created: 2015-09-15 11:00:00

Date modified: 2015-09-18 03:13:45

HSP70A-RBCS2_Hyg Expression Plasmid

Details

• Types: DnaRegion
• Roles: Project; engineered_region
• Sequences: BBa_K1857002_sequence (Version 1)

Description

This plasmid is designed to facilitate the easiest expression of proteins possible. It contains an HSP70A-RBCS2 fusion promoter attached to an RBCS2 intron, flanked by the iGEM prefix and suffix. There is an RBCS2 3' UTR after the suffix. Additionally, there is a complete Hygromycin B resistance cassette downstream, allowing for selection.

The suffix can be removed and replaced with the coding sequence of a heterologous protein. Furthermore, the entire fusion-promoter-intron sequence can be removed with the prefix and suffix and replaced by an alternate promoter to compare expression levels between Chlamydomonas promoters.

Notes

We amplified the pHSP70A-RBCS2-Intron region with the iGEM prefix/suffix added to the ends. We then amplified 3'UTR RBCS2 with PstI upstream' and SpeI downstream. The hygromycin B resistance cassette was then amplified with XbaI upstream and SpeI downstream. pSB1C3 was amplified with primers which removed ligated with digested pHSP70A-RBCS2-Intron and pSB1C3 for final assembly.

Source

The parts were adapted from pOpt_mVenus_Hyg.

Sequence Annotations

Sequence Annotation	Location	Component / Role(s)
pHSP70A	1,267	promoter, feature/promoter
pRBCS2	274,497	promoter, feature/promoter

Other Properties

• igem#status: Planning
• synbiohub#ownedBy: user/james
• synbiohub#ownedBy: user/myers
• synbiohub#topLevel: BBa_K1857002/1

Member of these Collections

• iGEM Parts Registry