Difference between revisions of "Part:BBa K3505023"
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<partinfo>BBa_K3505023 short</partinfo> | <partinfo>BBa_K3505023 short</partinfo> | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | + | FFAR2 is a naturally found eukaryotic GPCR protein that exhibits high affinity for SCFAs (acetic, propionic and butyric acid)(Kaemmerer, 2010) . After background searching through the NCBI database, we have identified the gene coding sequences needed for the designing of this gene construct. More specifically, a vasopressin receptor 2 segment has been attached to the C-terminal of the receptor, as it mediates recruitment of a TEV tagged b-arrestin-2, along with a TEV cleavage site(TCS). | |
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===Design Notes=== | ===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 | + | 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 GoldenBraid cloning. |
The CDS has position B2-B5. | The CDS has position B2-B5. | ||
<p style="text-align: center;"> | <p style="text-align: center;"> | ||
− | [[Image: T--Thessaly--GB-CCAT-GCTT.jpeg|900px|thumb|none|<i><b>Figure 1.</b>The overhangs of this part in the | + | [[Image: T--Thessaly--GB-CCAT-GCTT.jpeg|900px|thumb|none|<i><b>Figure 1.</b>The overhangs of this part in the GoldenBraid Grammar.</i>]] |
[[Image:T--Thessaly--FFAR2-snap.png|900px|thumb|none|<I><b>Figure 2.</b> The level 0 module : pupd2-FFAR2:V2tail:TCS (illustration from SnapGene)</i>]] | [[Image:T--Thessaly--FFAR2-snap.png|900px|thumb|none|<I><b>Figure 2.</b> The level 0 module : pupd2-FFAR2:V2tail:TCS (illustration from SnapGene)</i>]] | ||
===Verification of Cloning=== | ===Verification of Cloning=== | ||
− | [[File:T--Thessaly--FFAR2-ECFP-digestion.png|700px|thumb|none|<i><b>Fig. | + | [[File:T--Thessaly--FFAR2-ECFP-digestion.png|700px|thumb|none|<i><b>Fig.3:</b>(U=Uncut C=Cut) Restriction Digestion of FFAR2:V2tail:TCS with BamHI, Expected bands : 3204 bp, Positive Result:C2,C3</i>]] |
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+ | ===Source=== | ||
+ | Synthesized by IDT. | ||
===Sequence and Features=== | ===Sequence and Features=== | ||
<partinfo>BBa_K3505023 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3505023 SequenceAndFeatures</partinfo> | ||
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+ | ===References=== | ||
+ | *Kaemmerer, E., 2010. Fatty acid binding receptors in intestinal physiology and pathophysiology. World Journal of Gastrointestinal Pathophysiology, 1(5), p.147. |
Latest revision as of 00:28, 28 October 2020
FFAR2:V2tail:TCS GB compatible with B2-B3
Usage and Biology
FFAR2 is a naturally found eukaryotic GPCR protein that exhibits high affinity for SCFAs (acetic, propionic and butyric acid)(Kaemmerer, 2010) . After background searching through the NCBI database, we have identified the gene coding sequences needed for the designing of this gene construct. More specifically, a vasopressin receptor 2 segment has been attached to the C-terminal of the receptor, as it mediates recruitment of a TEV tagged b-arrestin-2, along with a TEV cleavage site(TCS).
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 GoldenBraid cloning. The CDS has position B2-B5.
Verification of Cloning
Source
Synthesized by IDT.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 284
Illegal PstI site found at 864 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 284
Illegal PstI site found at 864 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 106
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 284
Illegal PstI site found at 864 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 284
Illegal PstI site found at 864
Illegal NgoMIV site found at 364 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 560
Illegal SapI site found at 815
Illegal SapI.rc site found at 17
References
- Kaemmerer, E., 2010. Fatty acid binding receptors in intestinal physiology and pathophysiology. World Journal of Gastrointestinal Pathophysiology, 1(5), p.147.