Coding

Part:BBa_K3505024

Designed by: Asteria Tsapadikou, Emmanouil Stylianakis   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 15:09, 26 October 2020 by Asteriat (Talk | contribs)


B-arrestin:TEV Protease GB compatible with B2-B5

long


Usage and Biology

Design Notes

The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is presents in pUPD2 and has overhangs compatible for Golden Braid cloning. The CDS has position B2-B5.

File:T--Thessaly--GB-CCAT-GCTT.jpeg .jpeg
Figure 1.The overhangs of this part in the Golden Braid Grammar.


Figure 2. The level 0 module : pupd2-b-arrestin-TEV protease (illustration from SnapGene)

Verification of Cloning

Fig.1:(U=Uncut C=Cut) Restriction Digestion of b-arrestin-TEV protease with SlaI, Expected bands : 2014 bp + 892 bp + 760 bp + 394 bp, Positive Result: green arrows


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1611
    Illegal PstI site found at 303
    Illegal PstI site found at 390
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1611
    Illegal PstI site found at 303
    Illegal PstI site found at 390
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 203
    Illegal XhoI site found at 600
    Illegal XhoI site found at 994
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1611
    Illegal PstI site found at 303
    Illegal PstI site found at 390
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1611
    Illegal PstI site found at 303
    Illegal PstI site found at 390
    Illegal AgeI site found at 586
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1545
    Illegal SapI.rc site found at 1893

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