Composite

Part:BBa_K3505037

Designed by: Asteria Tsapadikou   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 14:34, 26 October 2020 by Asteriat (Talk | contribs) (→‎Sequence and Features)


pAndersonJ23115:RBS-TetR-terminator- pAndersonJ23115:tetO:RBS-LacI-terminator

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 cloned in omega1 vector BBa_K3505010 and has overhangs compatible for Golden Braid cloning.

Figure 2. The level B module of Tet-Off system : a1R:AndersonJ23115:RBS-TetR-Double terminator-a2:AndersonJ23115:TetO:RBS- LacI-Double terminator

Verification of Cloning

Fig.1:: (U=Uncut , C= Cut) Restriction digestion of omega1R-TetR-LacI (C1-C 4) with : �EcoRV +EcoRI(C1-C4) , Expected bands : 4823 + 465bp ,Positive result: C1,C2,C3,C4

Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 76
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
    Illegal NheI site found at 838
    Illegal NheI site found at 861
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 76
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 76
  • 1000
    COMPATIBLE WITH RFC[1000]
[edit]
Categories
Parameters
None