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>]]
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===Verification of Cloning===
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[[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>]]
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===Sequence and Features===
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<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.

Figure 2. The level B module of GPCR-Tango Module : a1R:ParaBAD:RBS-FFAR2:V2tail:TCS-Lac-Double terminator

Verification of Cloning

Fig.1:: (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)


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]

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1527
    Illegal PstI site found at 2107
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1527
    Illegal PstI site found at 2107
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1148
    Illegal BamHI site found at 1349
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1527
    Illegal PstI site found at 2107
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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