Difference between revisions of "Part:BBa K3505037"

Revision as of 14:35, 26 October 2020

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

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]

@@ Line 15: / Line 15: @@
 ===Verification of Cloning===
-[[File:T--Thessaly--omga1-tetoff-digestion.png|700px|thumb|none|<i><b>Fig.1:</b>: (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</i>]]
+[[File:T--Thessaly--omga1-tetoff-digestion.png|700px|thumb|none|<i><b>Fig.1:</b>: (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</i>]]
 ===Sequence and Features===
 <partinfo>BBa_K3505037 SequenceAndFeatures</partinfo>