Part:BBa_J70010

A BioScaffold Part (Uses PpiI), for offset see Part Design page

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

still in testing, uses PpiI (2) PacI (2) MabI (2), currently 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:

    TM=54.3               TM=53.8                 TM=58.6
 EcoRINotI     XbaI                                                            SpeI
 AATTCGCGGCCGCTTCTAGAG| gagagctggttcaccaggtt aattaattaattaaacctggtgaacgtggtctc ta (ctag) 
     G                                      |                                  at  gatc
gaattcgcggccgcttctagag                                                         ta  ctagtagcggccgctgcag
BioBrick Prefix         gagagctggttcaccaggtt aattaattaattaaacctggtgaacgtggtctc BioBrick Suffix                                          
                        J70010 sequence

Oligos for J70010.2 (3/17/09)
                                              
Forward 1.2 (TM=54.3): 5'-AATTCGCGGCCGCTTCTAGAG-3'         (this oligo corrects an error from version 1.1) 
Forward 2.2 (TM1=53.8, TM2=58.6): 5'-gagagctggttcaccaggttaattaattaattaaacctggtgaacgtggtctcta-3'
Reverse 2.2 (TM1=53.8,54.3): 5'-aacctggtgaaccagctctcCTCTAGAAGCGGCCGCG-3' (This oligo corrects a version 2.1 error)
Reverse 1.2: 5'-ctagtagagaccacgttcaccaggtttaattaattaatt-3'

Obsolete Versions:

Forward 1.1: AATTCGCGGCCGCTACTAGAG (bug)
Forward 2.1: gagagctggttcaccaggttaattaattaattaaacctggtgaacgtggtctcta
Reverse 1.1: CTAGTAGAgaccacgttcaccaggtttaattaattaatt
Reverse 2.1: aacctggtgaaccagctctcCTCTAGTAGCGGCCGCG (bug)

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 48