Part:BBa_K4195140

J23100-B0030-array(targeting pirA/B)-B0015

Biology

This part was designed as an element of typeⅠCRISPR/Cas system, in which the array contains several designed spacers to target pirA/B gene and repeats derived from Vibrio parahaemolyticus FOC_022 (1) that could be recognized and handled by Cas proteins.

Usage and design

The circuit of the basic part BBa_K4195023 was assembled into vector pUC57-simple to construct this composite part.

Characterization

1. Identification

When constructing this circuit, regular PCR and gene sequencing were used to verify that the plasmids were correct. Target bands (418 bp) can be observed at the position between 250 bp and 500 bp (Fig. 1).

Fig. 1 DNA gel electrophoresis of the regular PCR products of BBa_K4195140_pUC57-simple.

Reference

1. P. Baliga, M. Shekar, M. N. Venugopal, Investigation of direct repeats, spacers and proteins associated with clustered regularly interspaced short palindromic repeat (CRISPR) system of Vibrio parahaemolyticus. Mol. Genet. Genomics 294, 253-262 (2019).

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]