Part:BBa_J70210

A BioScaffold Part (Uses PpiI), similar to J70010 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)

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
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 25
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 49
  Illegal BsaI.rc site found at 1