Composite
CMV AID pA

Part:BBa_K929000

Designed by: Potsdam Bioware 2012 iGEM   Group: iGEM12_Potsdam_Bioware   (2012-09-16)
Revision as of 11:15, 25 September 2012 by ChristopherKressler (Talk | contribs)

AID with CMV promoter and hGH-polyadenylation signal sequence

CMV_wtAID_pA
UP12 PlasmidCMV wtAID pA.png
BioBrick Nr. BBa_K929000
RFC standard RFC 10
Requirement pSB1C3
Source existing parts:(BBa_K103001;BBa_I712004; BBa_K404108)
Submitted by [http://2012.igem.org/Team:Potsdam_Bioware Potsdam_Bioware2012]


UP12 PlasmidCMV wtAID pA2.png





















The BioBrick "AID with CMV promoter and hGH-polyadenylation signal sequence" (BBa_K929000) is an extended version of the existing AID BioBrick (BBa_K103001). It is built of 3 parts: CMV promoter (BBa_I712004), AID BBa_K103001)and hGH polyadenylation signal sequence (BBa_K404108).
AID:
AID is known to be responsible for somatic hypermutation and the class-switch recombination of immunoglobulin in B cells. This enzyme of 28 kDa originally occurs in B cells but does also show activity after transfection into CHO cells. AID induces the deamination of cytidine to uridine at actively transcribed single strand DNA. The replacement of cytidine by uridine leads to a mismatch during DNA replication and integrates a single base substitution predominantly in the immunoglobulin genes.
CMV promoter:
CMV is an immediate-early Cytomegalovirus promoter for high-level expression. The CMV promoter is commonly used due to its very strong activity and effectivity in a broad range of cell types. The BioBrick is therefore improved via addition of the strong promoter.
hGH polyadenylation signal sequence:
Polyadenylation is a significant part for the translation and stability of mRNA. In eukaryotes, it is part of the process that produces mature messenger RNA (mRNA) for translation. It, therefore, forms part of the larger process of gene expression. hGH terminator gives a signal to start polyadenylation in the translation process.


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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1661
    Illegal BsaI.rc site found at 740
    Illegal SapI site found at 841


[edit]
Categories
Parameters
None