Part:BBa_K4695110

pnbA-docscaB

This is a protein-coding composite part consisting of two protein domain-coding sequences with the DT4_TEV linker between them and a HisTag linker T5E_6H at the C-terminal end. Expressing this sequence results in a fusion protein capable of di-ester bond hydrolysis of phthalic acid esters (PAEs).This protein can also bind to corresponding cohesin domain.

Design Notes

This composite part consists of:

• PnbA coding sequence (BBa_K4695010)
• DT4_TEV linker (BBa_J435395)
• DocScaB coding sequence (BBa_K4695041)
• C-terminal T5E_6H HisTag linker (BBa_J435369)

Sequence is designed in a way that allows user to swap enzymatic and dockerin domains with their counterparts.
FLANKED 5' fusion site: AATG, 3' fusion site: GCTT.
Source sequences were optimized and checked for BsaI and SapI restriction sites using: http://genomes.urv.es/OPTIMIZER/

Source

PnbA:
Organism: Bacillus subtilis BJQ0005
GenBank page: https://www.ncbi.nlm.nih.gov/protein/QHH21706.1/
Article: https://doi.org/10.1016/j.envpol.2021.116461

DocScaB:
Organism: Acetivibrio cellulolyticus
Structure: https://www.rcsb.org/structure/4UYQ

Registry of standard biological parts

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1603
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1603
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1603
  Illegal BamHI site found at 534
  Illegal BamHI site found at 837
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1603
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1603
  Illegal NgoMIV site found at 603
  
• 1000
  COMPATIBLE WITH RFC[1000]