Part:BBa_K2230001

EmR/pSB1C3

Erythromycin Resistance Gene as a Selection Marker for Lactobacillus transformation

Erythromycin is a common selection marker for engineering lactic acid bacteria. And teams [http://2013.igem.org/Team:Uppsala Uppsala] in 2013 and [http://2016.igem.org/Team:TMMU_China TMMU_China] in 2016 were using it to select the engineered Lactobacillus with erythromycin resistant strains. However, the BioBrick parts containing the erythromycin resistance gene are currently unavailable. On the other hand, iGEM HQ is unable to service the request of plasmid carrying erythromycin resistance gene. Maybe iGEM HQ can’t manipulate erythromycin resistant bacteria so far. Therefore, we decided to clone the erythromycin resistance gene (EmR) by ourselves. We searched the vector carrying EmR and found an iGEM team HKUST in 2010 working with it. We got the pMG36e vector from the team and cloned the EmR gene out onto pSB1C3 (EmR/pSB1C3, BBa_K2230001). We also made RBS-EmR/pSB1C3 (BBa_K2230002) and RBS-EmR-TT/pSB1C3 (BBa_K2230003) by assembling RBS (B0034) and double terminator (B0015) parts, respectively. These will provide iGEM team this resistance gene as selection marker in the future.

Cloning

EmR (Erythromycin resistance gene) was amplified from pMG36e vector and cloned onto pSB1C3.

Reference

Genetic engineering techniques for lactic acid bacteria: construction of a stable shuttle vector and expression vector for β-glucuronidase. Biotechnol Lett. 2014;36(2):327-35.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]