Difference between revisions of "Part:BBa K3505012"

Revision as of 10:36, 26 October 2020

AndersonJ23115:RBS GB compatible with A1-B2

Constitutive Anderson BBa_J23115with RBS BBa_B0034

Fig.1:J23115 with RBS

Design Notes

The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 BBa_K3505007 and has overhangs compatible for Golden Braid cloning. This has position A1-B3.

Fig.2:The overhangs of this part in the Golden Braid Grammar

Fig.1:J23115 with RBS

Verification of cloning

Fig.1:((U=Uncut , C= Cut) Restriction digestion of AndersonJ23115 with : EcoRI (C1a & C2a),Expected bands : 2156 bp, XhoI (C1b & C2b),Expected bands : 1264 + 892 bp Positive result: The set of C1a and C1b (same sample)

Experimental Use and Experience

This part is used in BBa_K3505033 BBa_K3505035

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 11
Illegal NheI site found at 34
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 20: / Line 20: @@
 [[File:T--Thessaly--GB-GGAG-AATG.jpeg|700px|thumb|none|<i><b>Fig.2:</b>The overhangs of this part in the Golden Braid Grammar</i>]]
-===Verification of cloning===
+[[File:T--Thessaly--AndersonRBS-snap.png|700px|thumb|none|<i><b>Fig.1:</b>J23115 with RBS</i>]]
+===Verification of cloning===
+[[File:T--Thessaly--Anderson--digestion.png|700px|thumb|none|<i><b>Fig.1:</b>((U=Uncut , C= Cut) Restriction digestion of AndersonJ23115 with :
+EcoRI (C1a & C2a),Expected bands : 2156 bp,
+XhoI (C1b & C2b),Expected bands : 1264 + 892 bp
+Positive result: The set of C1a and C1b (same sample) </i>]]
 ===Experimental Use and Experience===