Part:BBa_K578003

RSR Array Segment (Listeria innocua CLIP11262)

Note: this brick is intended to function with BBa_K578002, tracrRNA (not yet submitted), and Cas9 (not yet submitted). It is a functional unit of a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) array based around Listeria Innocua CLIP11262. The sequence of the part is composed of a Repeat, Spacer, Repeat. (Targeted to sequence with correct protospacer and 'NGG; Protospacer-adjacent motif), followed by a palindromic repeat. After the aforementioned sequences there is a pseudospacer with the standard BioBrick suffix (with SpeI and PstI Sites). The suffix allows for parts (eg.Repeat.Spacer Repeat, RSR, formated BioBricks) to be added onto the distal end of the CRISPR array.

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]

Confirmation of Part Length

PCR of BBa_K578003 using Primers VF2 (BBa_G00100) and VFR (BBa_G00101) from pSB1C3. The part is 135bp plus 42bp for the Standard BioBrick prefix and suffix. Note: The primers add approximatellly 200bp to the BioBrick's size, thus BBa_K578003 should appear at ~377bp.

Use of this BioBrick

This part is intended for use with BBa_K578002 to build up a repeat/spacer array that targets GFP expression.