Difference between revisions of "Part:BBa K228867"

Revision as of 04:38, 20 October 2009

pSB1A2-lacZ screening plasmid

This part is an improved version of pSB1A2, which lacks the NotI in between SpeI and PstI site. A constitutive lacZ alpha generator is placed in the original NotI site. Usually, rbs parts and promoter parts which is relatively small and has a low production after gel purification can be placed in this backbone. This improved plasmid can support blue white screening. Because the original parts on this plasmid is small parts, it is digested to be a vector. By cutting the plasmid with SpeI and PstI, the lacZ generator is removed. Although false positive can be created by religation of singal digestion product, it can be distinguished from the correct colonies by its color. Correct colonies are white while false positive ones are blue.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Plasmid lacks a suffix.
Illegal SpeI site found at 2
Illegal PstI site found at 303
12
INCOMPATIBLE WITH RFC[12]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2345
Illegal NheI site found at 14
Illegal NheI site found at 37
Illegal SpeI site found at 2
Illegal PstI site found at 303
Illegal NotI site found at 2351
21
INCOMPATIBLE WITH RFC[21]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2345
23
INCOMPATIBLE WITH RFC[23]
Illegal prefix found at 2345
Plasmid lacks a suffix.
Illegal SpeI site found at 2
Illegal PstI site found at 303
25
INCOMPATIBLE WITH RFC[25]
Illegal prefix found at 2345
Plasmid lacks a suffix.
Illegal XbaI site found at 2360
Illegal SpeI site found at 2
Illegal PstI site found at 303
1000
INCOMPATIBLE WITH RFC[1000]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI.rc site found at 1384

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K228867 short</partinfo>
-T7 polymerase with an amber mutation coding gene is constructed after AraC protein(reversed sequence) and Pbad promoter which would be triggered by Arabinose. T7 polymerase can work well in the exsitence of SupD tRNA. Besides, the expression of the T7 polymerase can be regulated by RBS.
+This part is an improved version of pSB1A2, which lacks the NotI in between SpeI and PstI site. A constitutive lacZ alpha generator is placed in the original NotI site. Usually, rbs parts and promoter parts which is relatively small and has a low production after gel purification can be placed in this backbone. This improved plasmid can support blue white screening. Because the original parts on this plasmid is small parts, it is digested to be a vector. By cutting the plasmid with SpeI and PstI, the lacZ generator is removed. Although false positive can be created by religation of singal digestion product, it can be distinguished from the correct colonies by its color. Correct colonies are white while false positive ones are blue.
 <!-- Add more about the biology of this part here