Part:BBa_K1041000

RFP Coding Device

Mutagenesis was used to add a Nde1 site at the start of the RFP coding region of the biobrick BBa_J04450. This enables a restriction digest with Nde1 to be performed allowing the RFP coding region to be excised from the plasmid.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 781
  Illegal AgeI site found at 893
  
• 1000
  COMPATIBLE WITH RFC[1000]

Composite part of the following BioBricks:

LacI R0010	B0034	mRFP1 E1010	B0015

• BBa_R0010: Promoter (lacI regulated)
  
• BBa_B0034: RBS (Elowitz 1999)
  
• BBa_E1010: Red Fluorescent Protein from Discosoma striata
  
• BBa_B0015: Double Terminator
  
• BBa_B0010: Transcriptional terminator with stem loop
  
• BBa_B0012: Transcriptional terminator for E.coliRNA polymerase
  

Characterisation

Fig 1:(Left) BBa_K1041000 and (right) BBa_J04450 visualized under non-UV lightbox

Both parts BBa_K1041000 and BBa_J04450 have colonies that appear red under natural light fig.1. The addition of a NdeI site has not effected the ability for the RFP gene to be transcribed.