Difference between revisions of "Part:BBa K1467300"

Latest revision as of 10:29, 9 September 2015

Golden Gate Module Flipper for 3'UTR+Terminator parts

This part is a "GoldenGate Flipper" that can be used to convert 3U+Ter (3' untranslated region and terminator)modules from the GoldenGate MoClo Assembly Standard into standard BioBricks. It consists of the RFP reporter (from BBa_J04450) flanked by an inverted pair of BsaI recognition sequences. GoldenGate MoClo parts can be flipped into BrioBrick parts in a one-pot, one-step, digestion-ligation GoldenGate cloning reaction. The image below shows the sequences inserted (in orange) between the RFP and the BioBrick prefix and suffix that enable the flipper to accept MoClo 3U+Ter parts, which begin GCTT and end CGCT.

The colonies containing this part are clearly red in color under natural light after about 18 hours. Smaller colonies are visibly red under UV. The RFP part does not contain a degradation tag and the RBS is strong.When cloning is successful the colonies become white as the RFP sequence is replaced by the new part. After the reaction is complete, no part of the BsaI recognition sequence will remain between the BioBrick suffix and the part.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 793
Illegal AgeI site found at 905
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1082
Illegal BsaI.rc site found at 6

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1467300 short</partinfo>
-This part is a "GoldenGate Flipper" that can be used to convert 3U+Ter (3' untranslated region and terminator) modules from the GoldenGate MoClo Assembly Standard into standard BioBricks. Flanking the RFP is an inverted pair of BsaI recognition sequences either side of the original RFP reporter part. The flipper reaction is a one-pot, one-step, digestion-ligation GoldenGate cloning reaction. The image below shows the sequences inserted (in orange) between the RFP and the BioBrick prefix and suffix that enable the flipper to accept MoClo CDS1 parts, which begin GCTT and end CGCT, in a one-step digestion-ligation Golden Gate cloning reaction.
+This part is a "GoldenGate Flipper" that can be used to convert 3U+Ter (3' untranslated region and terminator)modules from the GoldenGate MoClo Assembly Standard into standard BioBricks. It consists of the RFP reporter (from BBa_J04450) flanked by an inverted pair of BsaI recognition sequences. GoldenGate MoClo parts can be flipped into BrioBrick parts in a one-pot, one-step, digestion-ligation GoldenGate cloning reaction. The image below shows the sequences inserted (in orange) between the RFP and the BioBrick prefix and suffix that enable the flipper to accept MoClo 3U+Ter parts, which begin GCTT and end CGCT.
 [[Image:terflipper.jpg|600px|]]
@@ Line 9: / Line 9: @@
 The colonies containing this part  are clearly red in color under natural light after about 18 hours. Smaller colonies are visibly red under UV. The RFP part does not contain a degradation tag and the RBS is strong.When cloning is successful the colonies become white as the RFP sequence is replaced by the new part. After the reaction is complete, no part of the BsaI recognition sequence will remain between the BioBrick suffix and the part.
-The colonies containing this part  are clearly red in color under natural light after about 18 hours. Smaller colonies are visibly red under UV. The RFP part does not contain a degradation tag and the RBS is strong.When cloning is successful the colonies become white as the RFP sequence is replaced by the new part. After the reaction is complete, no part of the BsaI recognition sequence will remain between the BioBrick suffix and the part.