Difference between revisions of "Part:BBa J119022"

Revision as of 14:49, 20 August 2012

HPP Bsa J23100 promoter in pSB1A2

J119022 can be used with either Part:BBa_J119044 or Part:BBa_J100091 for Golden Gate Assembly (GGA) to move promoter Part:BBa_J23100 to replace the transcriptional terminator (TT). J119022 is an ideal way to start learning how easy it is to swap two parts using [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate GGA.] You can follow a [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate_Assembly_protocol complete protocol for use in teaching labs.]

J119022 contains a BsaI-flanked Part:BBa_J23100 promoter in pSB1A8 (Part:BBa_J119043) for BsaI/Ligase insertion into Part:BBa_J119044 or Part:BBa_J100091.

The Part:BBa_J23100 promoter is designed for simultaneous BsaI digestion and ligation into Part:BBa_J119044 or Part:BBa_J100091 as a positive control for use in Golden Gate Assembly. J119022 is the first promoter to be use as part of the [http://gcat.davidson.edu/RFP/ Registry of Functional Promoters].

J119022 is in pSB1A8 Part:BBa_J119043 with a BsaI site on the left cutting to the right, with sticky end CGAC being freed up, followed by Part:BBa_J23100 promoter, followed up by a different sticky end of GCGG which is freed up by the following BsaI site which cuts to the left. The fragment is 57bp long after digestion. It should then be ligated into (Part:BBa_J119044) or Part:BBa_J100091.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 15
Illegal SpeI site found at 80
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 39
Illegal NheI site found at 62
Illegal SpeI site found at 80
Illegal NotI site found at 6
Illegal NotI site found at 87
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 15
Illegal SpeI site found at 80
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 15
Illegal SpeI site found at 80
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 22
Illegal BsaI.rc site found at 73

@@ Line 4: / Line 4: @@
 J119022 can be used with either [[Part:BBa_J119044]] or [[Part:BBa_J100091]] for Golden Gate Assembly (GGA) to move promoter [[Part:BBa_J23100]] to replace the transcriptional terminator (TT). '''J119022''' is an ideal way to start learning how easy it is to swap two parts using [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate GGA.] You can follow a [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate_Assembly_protocol complete protocol for use in teaching labs.] <br><br>
-'''J119022'''  contains a BsaI-flanked [[Part:BBa_J23100]] promoter in pSB1A8 ([[Part:BBa_J119043]]) for BsaI/Ligase insertion into ([[Part:BBa_J119044]]), a modified version of [[Part:BBa_J100028]].
+'''J119022'''  contains a BsaI-flanked [[Part:BBa_J23100]] promoter in pSB1A8 ([[Part:BBa_J119043]]) for BsaI/Ligase insertion into [[Part:BBa_J119044]] or [[Part:BBa_J100091]].
-This is the [[Part:BBa_J23100]] promoter fragment designed for BsaI digestion and ligation into [[Part:BBa_J119044]] as a positive control for use in Golden Gate Assembly. J119022 is the first promoter to be use as part of the [http://gcat.davidson.edu/RFP/ Registry of Functional Promoters].
+The [[Part:BBa_J23100]] promoter is designed for simultaneous BsaI digestion and ligation into [[Part:BBa_J119044]] or [[Part:BBa_J100091]] as a positive control for use in Golden Gate Assembly. J119022 is the first promoter to be use as part of the [http://gcat.davidson.edu/RFP/ Registry of Functional Promoters].
-'''J119022''' is in pSB1A8 [[Part:BBa_J119043]] with a BsaI site on the left reading right, with word CGAC being freed up, followed by [[Part:BBa_J23100]] promoter, followed up by word GCGG which is freed up by the following BsaI site which reads to the left.  The fragment is 57nt long after digestion.  It should then be ligated into ([[Part:BBa_J119044]]) or a vector designed to accept the sticky end "Words" noted as "4NT Word" on the fragment diagram. (Requires a complement sticky end to be compatible).
+'''J119022''' is in pSB1A8 [[Part:BBa_J119043]] with a BsaI site on the left cutting to the right, with sticky end CGAC being freed up, followed by [[Part:BBa_J23100]] promoter, followed up by a different sticky end of GCGG which is freed up by the following BsaI site which cuts to the left.  The fragment is 57bp long after digestion.  It should then be ligated into ([[Part:BBa_J119044]]) or [[Part:BBa_J100091]].