Team:TECHNION-ISRAEL/Basic Part

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Basic Parts

Basic Parts




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
My First Website

Department of Biotechnology & Food Engineering
Technion – Israel Institute of Technology
Haifa 32000, Israel

    • igem.technion.2017@gmail.com