Part:BBa_J70030

A BioScaffold Part (Uses PpiI), Protein Tail Remover see Part Design Page

When placed in a BioBrick assembly and cut with the restriction enzyme PpiI, this part will direct the restriction enzymes to cut outside the BioBrick scar

still in testing, uses restriction enzyme sites PpiI (2) PacI (2) MabI (2), will place in pSB1AK3 vector

See part design page or BBFRFC15, for details about use

This part can be made with the following oligos, which can be annealed and placed into an EcoRI/SpeI cut vector:

Version 1.1 Oligos (These oligos were made properly but did not produce transformants in first try.) 

 EcoRINotI     XbaI                                                         SpeI
 AATTCGCGGCCGCTTCT | AGAG CTGGTTCACCTGGTTAATTAATTAATT  AAACCAGGTGAACGTGGTCTCTA (ctag)
     GCG                                              |                     AT  GATC
gaattcgcggccgcttct   agag                                                   ta  ctagtagcggccgctgcag
BioBricks Prefix                                                            BioBricks Suffix
                          ctggttcacctggttaattaattaatt  aaaccaggtgaacgtggtctc
                          J70030 sequence 
These have melting temperatures around 55 degrees Celsius. 

Forward 1.1: AATTCGCGGCCGCTTCT
Forward 2.1: AGAGCTGGTTCACCTGGTTAATTAATTAATTAAACCAGGTGAACGTGGTCTCTA
Reverse 1.1: AATTAATTAATTAACCAGGTGAACCAGCTCTAGAAGCGGCCGCG
Reverse 2.1: ctagTAGAGACCACGTTCACCTGGTTT

This part is currently being placed into the vector pSB2K4, which will allow hierarchical manipulations of the part.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 43