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Revision as of 01:58, 2 November 2017
Basic Parts
![](https://static.igem.org/mediawiki/2017/8/81/T--TECHNION-ISRAEL--basic_part.png)
We designed four new basic parts that were used in our project:
Mutated EF-1a promoter (BBa_K2520023) is a strong, constitutive, promoter in mammalian cells. It is particularly useful in cells that do not express proteins with other common promoters (such as CMV). This promoter is commonly used in stem cells as it is one of the few promoters that prompt high levels of protein expression in these predominantly dormant cells.
This promoter is our favorite part. Promoters are clearly very important in iGEM. Synthetic biology depends on strong and consistent expression of recombinant proteins. The EF-1a promoter was used by both teams who won “best therapeutics project” in 2016 (Tel-Hai, SYSU MEDICINE), but neither submitted it as a part since the promoter contains two forbidden restriction sites within its sequence. As a side project, we set out to create a mutated version of the original (WT) EF-1a so that it may serve future iGEM teams. We mutated the WT EF-1a at two points (base pair 319 and 824 were changed from C to T), thus eliminating the forbidden restriction sites. We characterized and tested our new promoter for functionality and compared it with the WT EF-1a (Figure 1 at the part's page). In closing, we have added a new and valuable promoter to the iGEM parts registry that we believe will serve future teams seeking to work with mammalian cells.
Secrecon (BBa_K2520002) is an artificial secretory signal, which leads to high levels of protein secretion and membrane expression. Secrecon was found to be the most powerful and consistent human secretion signal sequence (more than Igk, for example).
B7-1 (BBa_K2520000) is a trans-membrane domain protein with a cytoplasmic tail. It enhances transport from the endoplasmic reticulum and allows accumulation of chimeric proteins on the plasma membrane. It has been shown to allow for the most consistent and long lasting membrane expression in human cells (more than PDGFR, for example).
Rat anti-mouse IgM scFv (BBa_K2520001) is a synthetic version of an anti-mouse IgM antibody. It binds the constant heavy region of all mouse antibodies with the IgM isotype. This recombinant protein can be used to bind primary (IgM) antibodies from murine source.
Name | Type | Description | Designer | Length | |||
---|---|---|---|---|---|---|---|
BBa_K2520000 | Protein_Domain | B7-1 Murine Trans Membrane Domain | Dana Kadosh, Noa Eden | 213 | |||
BBa_K2520001 | Coding | Single Chain Fragment Variable (scFv) of rat anti-murine IgM antibody | Noa Eden, Dana Kadosh | 699 | |||
BBa_K2520002 | Coding | Secrecon | Noa Eden, Dana Kadosh | 63 | |||
favorite | BBa_K2520023 | Regulatory | EF1a promoter | Dana Kadosh, Noa Eden | 1254 | ||
BBa_K2520024 | Signalling | IgK- chain leader sequence | Noa Eden | 63 | |||
BBa_K2520025 | Regulatory | TRE promoter | Noa Eden | 317 | |||
BBa_K2520026 | Protein_Domain | platelet derived growth factor receptor (PDGFR) | Noa Eden | 147 | |||
BBa_K2520027 | Coding | Pro-insulin | Dana Kadosh | 810 | |||
BBa_K2520028 | Signalling | IgK- chain leader sequence | Noa Eden | 63 | |||
BBa_K2520029 | Signalling | IgK- chain leader sequence | Noa Eden | 63 | |||
BBa_K2520030 | Tag | HA Tag | Dana Kadosh | 27 | |||
BBa_K2520031 | Tag | Myc Tag | Dana Kadosh | 30 | |||
BBa_K2520032 | Tag | 6 x His Tag | Dana Kadosh | 16 | |||
BBa_K2520033 | Protein_Domain | P2A | Noa Eden | 66 | |||
BBa_K2520036 | Protein_Domain | PDGFR | Noa Eden | 147 | |||
BBa_K2520037 | Protein_Domain | PDGFR | Noa Eden | 147 | |||
BBa_K2520038 | Protein_Domain | MOG1 | Noa Eden | 63 | |||
BBa_K2520039 | Protein_Domain | MOG 2 | Dana Kadosh | 63 | |||
BBa_K2520040 | Protein_Domain | MOG 3 | Dana Kadosh | 42 | |||
BBa_K2520041 | Protein_Domain | Peanut epitope 1 - Arah2 / Arah6 | Dana Kadosh | 18 | |||
BBa_K2520042 | Protein_Domain | Peanut epitope 3 - Arah2 / Arah6 | Dana Kadosh | 18 | |||
BBa_K2520043 | Protein_Domain | Celiac epitope 1 | Dana Kadosh | 27 | |||
BBa_K2520044 | Protein_Domain | Celiac epitope 3 | Dana Kadosh | 27 | |||
BBa_K2520045 | Protein_Domain | Bee venom PLA epitope 1 | Dana Kadosh | 36 | |||
BBa_K2520046 | Protein_Domain | Bee venom PLA epitope 3 | Dana Kadosh | 54 |
![](https://static.igem.org/mediawiki/2017/f/f9/T--TECHNION-ISRAEL--newUpAB.png)