Part:BBa_K2669002

AmilCP: Codon Optimized basic part

BBa_K2669002 is a codon optimized AmilCP original native part Part:BBa_K592009. This sequence is codon optimized with the IDT optimization tool, and was optimized for E. coli K12. The native blue chromoprotein AmilCP is derived from the coral Acropora milleporais. The chromoprotein naturally exhibits a strong blue color when expressed. The color can be observed in both LB and agar culture and is visible with the naked eye. This part can be used as an indicator or as a reporter for various kinds of biological research.

Figure 1:Transformation plate of the IDT optimized AmilCP sequence.

Contribution: Guelph 2019

This year iGEM Guelph was working with BBa_K2669002 as a reporter protein. When working with this chromoprotein in E.coli BL21(DE3) under the control of BBa_K3189001, we found that stronger colour was produced at lower temperatures compared to higher temperatures. In Figure 2 pigment intensity was compared between two temperatures, 25°C and 37°C. It is clear form these images that at 25°C the colonies have a darker colour compared to colonies grown at 37°C. This is evidence that AmilCP produced from BBa_K2669002 is more stable at 25°C compared to 37°C thus allowing it to build up in the cells longer before degradation resulting in darker pigmentation of the colonies.


Figure 2: BBa_K2669002 expression at 25°C and 37°C. Expression was carried out in E.coli BL21(DE3) on LB agar supplemented with 100 ug/mL ampicillin, 100 mg/mL tryptophan, 1mM IPTG, and 50 ng/mL tetracycline.

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]