Part:BBa_K2657001

Tetracycline Resistance PhytoBrick

BBa_K2657001 is a PhytoBrick containing a Tetracycline resistance gene. It uses the Universal Acceptor (modified pSB1C3) as its backbone. Its overhangs make it a promoter-CDS part. It can be digested via BsaI.

Functionality

The functionality of this tetracycline resistance gene was demonstrated by the growth of electrocompetent E. coli. The TetR was used in a BsaI golden gate assembly reaction to create a self-replicating plasmid containing a ColE1/pMBI origin of replication as well as promoter and terminator sequences. genes encoding GFP and resistance to kanamycin. The assembly was transformed via electroporation into TOP10 E. Coli . The Mu Free Donor was then conjugated with Serratia marcescens . The conjugation was deemed successful by the organism's ability to express GFP and grow on a kanamycin plate.

Fig 1. Growth of electrocompetent E. coli with the TetR. The plates on the left show bacterial growth on tetracycline-containing media. Since the TOP10EC E. coli have no natural resistance to tetracycline, it can be inferred that they have been given resistance via the part electroporated into them.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 158
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 304
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 330
  Illegal NgoMIV site found at 698
  Illegal NgoMIV site found at 858
  Illegal AgeI site found at 1
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4