Plasmid
pSB1AT3X

Part:BBa_J18906

Designed by: Raik Gruenberg   Group: Affiliates   (2008-08-11)


pSB1AT3X Freiburg Fusion -> BioFusion conversion plasmid

pSB1AK3X shares the same backbone as pSB1AK3 but uses modified prefix and suffix sequences (that do not formally adhere to any BioBrick standard):

5' GAATTC GCGGCCGC T TCTAGA GCCGGC

  EcoRI    NotI     XbaI   NgoMIV
...part...
ACCGGT ACTAGT A GCGGCCG CTGCAG 3'

AgeI   SpeI     NotI    PstI 

These prefix / suffix sequences are designed to translate protein parts from the proposed Fusion (aka Freiburg) format to the older BioFusion format from the Silver lab. Construction vectors with this modified Freiburg prefix / suffix could bring a protein part in frame with BioFusion parts and remove the STOP between the AgeI and SpeI site. Restriction / ligation with AgeI + NaeI can (theoretically) transfer Expression parts into this conversion vector which can then be used for normal BioFusion cloning (at the cost of adding a T G before and after the part). NaeI is an isoschizomer to NgoMIV but generates blunt ends which should allow for a directional transfer.

See [http://openwetware.org/wiki/The_BioBricks_Foundation:Standards/Technical/Formats BB formats].

pSB1AK3X is a high copy number plasmid carrying ampicillin and tetracyclin resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell). For the version of this plasmid with a CcdB insert, see BBa_P1010.

The given sequence starts with the modified suffix and ends with the modified prefix. The complete vector sequence is therefore obtained from: part + vector.

See also:


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 3430
    Illegal suffix found in sequence at 6
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3430
    Illegal NheI site found at 1275
    Illegal SpeI site found at 7
    Illegal PstI site found at 21
    Illegal NotI site found at 14
    Illegal NotI site found at 3436
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3430
    Illegal BamHI site found at 1421
    Illegal XhoI site found at 1040
    Illegal XhoI site found at 2323
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 3430
    Illegal suffix found in sequence at 7
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 3430
    Illegal XbaI site found at 3445
    Illegal SpeI site found at 7
    Illegal PstI site found at 21
    Illegal NgoMIV site found at 3451
    Illegal AgeI site found at 1
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2469


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