Difference between revisions of "Part:BBa K3505025"
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[[Image:T--Thessaly--FFAR2-VTAIL-PHOT.png|900px|thumb|none|<I><b>Figure 2.</b> The level B module of GPCR-Tango Module : a1R:ParaBAD:RBS-FFAR2:V2tail:TCS-Lac-Double terminator </i>]] | [[Image:T--Thessaly--FFAR2-VTAIL-PHOT.png|900px|thumb|none|<I><b>Figure 2.</b> The level B module of GPCR-Tango Module : a1R:ParaBAD:RBS-FFAR2:V2tail:TCS-Lac-Double terminator </i>]] | ||
+ | |||
+ | ===Verification of Cloning=== | ||
+ | [[File:T--Thessaly--FFAR2-LACI-digestion.png|700px|thumb|none|<i><b>Fig.1:</b>: (U=Uncut , C= Cut) Restriction digestion a1R:ParaBAD:RBS-FFAR2:V2tail:TCS-Lac-Double terminato (C1a-C4b) with : BamHI(C1a-C4a) , Expected bands : 2847+2225 bp , EcoRV (C2a-C2b) ,Expected bands : 3587 bp + 2845 bp, Positive result: C1,C2,C3,C3 (C1a and C1b is the same sample etc)</i>]] | ||
+ | |||
+ | |||
+ | ===Sequence and Features=== | ||
+ | <partinfo>BBa_K3505037 SequenceAndFeatures</partinfo> | ||
===Sequence and Features=== | ===Sequence and Features=== | ||
<partinfo>BBa_K3505025 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3505025 SequenceAndFeatures</partinfo> |
Revision as of 16:46, 26 October 2020
ParaBAD:RBS-FFAR2:AVPR2 tail:TCS-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 alpha1R vector BBa_K3505008 and has overhangs compatible for Golden Braid cloning.
Verification of Cloning
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 76
- 12INCOMPATIBLE 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 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 76
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 76
- 1000COMPATIBLE WITH RFC[1000]
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1527
Illegal PstI site found at 2107 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1527
Illegal PstI site found at 2107 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1148
Illegal BamHI site found at 1349 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1527
Illegal PstI site found at 2107 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1527
Illegal PstI site found at 2107
Illegal NgoMIV site found at 1607
Illegal AgeI site found at 983 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 965
Illegal SapI site found at 1803
Illegal SapI site found at 2058
Illegal SapI.rc site found at 1260