Part:BBa_K1114702

BioBrick adaptation of MoClo Level 1 Reporter

This is a BioBrick adaptation of a Level 1 MoClo device that constitutively expresses RFP. The MoClo version of this device is BBa_K1114500.

This transcriptional unit contains the MoClo Level 0 parts J23104_AB, BCD1_BC, E0040m_CD, and B0015_DE within the pSB1C3 backbone. In the given sequence the internal fusion sites are incorrectly duplicated. The fusion site letters refer to 4bp fusion sites: A = GGAG; B = TACT; C = AATG; D = AGGT; E = GCTT; F = CGCT; G = TGCC; H = ACTA. See Level 0 pages for further information.

This device was converted into a BioBrick standard device using PCR to simultaneously remove the MoClo 5' and 3' sequences and insert the BioBrick 5' and 3' ends in their place. The PCR product was purified and digested with XbaI and PstI, and ligated into pSB1C3 (which was also cut with XbaI and PstI).

This part contains a bicistronic design (BCD) element as the 5' UTR regulatory part from Mutalik et al., 2013. From the library of BCDs, this devices contains a MoClo version of BCD1 (BioFAB # apFAB681) and we obtained the sequence data for this part from the BioFAB. The MoClo version of BCD1 is BBa_K1114106.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 11
  Illegal NheI site found at 34
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 694
  Illegal AgeI site found at 806
  
• 1000
  COMPATIBLE WITH RFC[1000]