Part:BBa_K4103005

catA9

CatA9 chloramphenicol (acetyl transferase) resistance fragment from pHT43.

Biology

Selectable markers are genes that confer the ability to grow in presence of toxic compounds[1]. The CatA9 gene produces a acetyl transferase enzyme that confers resistance to the antibiotic chloramphenicol.

Usage

Our team used the CatA9 biobrick as a selection marker to select our B. subtilis colonies that successful receveid our plasmid.

Name: catA9 BsaI Prefix: AAGG BsaI suffix: ATGA

This part contains the full length catA9 chloramphenicol (acetyl transferase) resistance fragment from pHT43 [1]. Includes inverted BtgzI overhangs outside of BsaI overhangs to flip gene orientation if needed.

NCBI Blast of promoter region: identical to Bacillus subtilis strain 75 chromosome 330510-330798 SubtiWiki genome search of this region: encodes for a portion of Idh (inositol dehydrogenase: https://www.uniprot.org/uniprot/P39126) and lctP (L-lactate permease: https://www.uniprot.org/uniprot/P55910) - which seems wrong (perhaps because of the strain differences from str. 75 & 168).

Protein Blast of CDS region: Chloramphenicol acetyltransferase from Streptococcus pneumoniae (also gram +ve) Promoter??

Might want to add a terminator at the end of the gene later on.

Note: added back the BsmBI site (3' sequence tctcata (rev) after terminator amino acid to reflect the exact fragment in pHT43) The full catA9 sequence reflect exactly the catA9 sequence in pHT43

References

[1]Jones HD. GENETIC MODIFICATION | Transformation, General Principles. Encyclopedia of Applied Plant Sciences. Published online 2003:377-382. doi:10.1016/b0-12-227050-9/00197-6 ‌ 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]