Difference between revisions of "Part:BBa K3505024"

Revision as of 15:11, 26 October 2020

B-arrestin:TEV Protease GB compatible with B2-B5

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 presents in pUPD2 and has overhangs compatible for Golden Braid cloning. The CDS has position B2-B5.

Figure 1.The overhangs of this part in the Golden Braid Grammar.

Figure 2. The level 0 module : pupd2-b-arrestin-TEV protease (illustration from SnapGene)

Verification of Cloning

Fig.1:(U=Uncut C=Cut) Restriction Digestion of b-arrestin-TEV protease with SlaI, Expected bands : 2014 bp + 892 bp + 760 bp + 394 bp, Positive Result: green arrows

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal SpeI site found at 1611
Illegal PstI site found at 303
Illegal PstI site found at 390
12
INCOMPATIBLE WITH RFC[12]
Illegal SpeI site found at 1611
Illegal PstI site found at 303
Illegal PstI site found at 390
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 203
Illegal XhoI site found at 600
Illegal XhoI site found at 994
23
INCOMPATIBLE WITH RFC[23]
Illegal SpeI site found at 1611
Illegal PstI site found at 303
Illegal PstI site found at 390
25
INCOMPATIBLE WITH RFC[25]
Illegal SpeI site found at 1611
Illegal PstI site found at 303
Illegal PstI site found at 390
Illegal AgeI site found at 586
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 1545
Illegal SapI.rc site found at 1893

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	<p style="text-align: center;">		<p style="text-align: center;">
−	[[Image:T--Thessaly--GB-CCAT-GCTT~~.jpeg~~ .jpeg\|900px\|thumb\|none\|<i><b>Figure 1.</b>The overhangs of this part in the Golden Braid Grammar.</i>]]	+	[[Image:T--Thessaly--GB-CCAT-GCTT.jpeg\|900px\|thumb\|none\|<i><b>Figure 1.</b>The overhangs of this part in the Golden Braid Grammar.</i>]]