Difference between revisions of "Part:BBa K3505044"

Revision as of 07:17, 27 October 2020

pAndersonJ23115:TetO:RBS-eCFP -terminator eCFP BBa_K3505020uder control of a constitutive promoter (andersonJ23115 with a tet operator) BBa_K2924014.

Fig.1:The Level A construct AndersonJ23115:TetO:rbs-eCFP-Terminator

Usage and Biology

This Trancriscription Unit (TU) is continuesly activated exressing the eCFP protein as a reporter. The Lac operator that is downsteam the anderson exists for the lac regulated inhibition.

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 a1R BBa_K3505008 and has overhangs compatible for Golden Braid cloning.

Verification of cloning

Fig.2:(U=Uncut C=Cut) (U=Uncut , C= Cut) Restriction digestion of TE: AndersonJ23115:Tet0-ECFP-double terminator(C1-C 4) with : EcoRV(C1-C2) , Expected bands : 2847+ 954 bp ,Positive result: C1,C2

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 13: / Line 13: @@
 ===Verification of cloning===
-[[File:T--Thessaly--TET0-ECFP-digestion.png|600px|thumb|none|<i><b>Fig.2:</b>(U=Uncut C=Cut) (U=Uncut , C= Cut) Restriction digestion of TE: AndersonJ23115:Tet0-EGFP-double terminator(C1-C 4) with : EcoRV(C1-C2) , Expected bands : 2847+ 954 bp ,Positive result: C1,C2</i>]]
+[[File:T--Thessaly--TET0-ECFP-digestion.png|600px|thumb|none|<i><b>Fig.2:</b>(U=Uncut C=Cut) (U=Uncut , C= Cut) Restriction digestion of TE: AndersonJ23115:Tet0-ECFP-double terminator(C1-C 4) with : EcoRV(C1-C2) , Expected bands : 2847+ 954 bp ,Positive result: C1,C2</i>]]
 ===Sequence and Features===
 <partinfo>BBa_K3505044 SequenceAndFeatures</partinfo>