Difference between revisions of "Part:BBa K635002"

 
 
(One intermediate revision by the same user not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K635002 short</partinfo>
 
<partinfo>BBa_K635002 short</partinfo>
Line 6: Line 5:
  
 
Yeast transformants can be selected by using a uracil drop out medium and E.coli transformants may be selected using ampicillin resistance.  
 
Yeast transformants can be selected by using a uracil drop out medium and E.coli transformants may be selected using ampicillin resistance.  
 +
 +
 +
 +
----
 +
 +
 +
 +
[[Image:Vgem2011-yeast-transformants.jpg|300px|left]] [[Image:Vgem2011-ecoli-transformants.jpg|300px|left]]
 +
 +
 +
 +
 +
 +
''S. cerevisiae'' (left)
 +
Colonies shown are successful yeast transformants using our backbone and BBa_J04450 (which cannot be be expressed as assembled in yeast).  Colonies were selected using a medium lacking uracil.  
 +
 +
 +
''E. coli'' (right)
 +
Red transformants indicate successful assembly of the backbone with the registry part BBa_J04450, which contains an RFP reporter protein.  The bacterial colonies were selected using ampicillin resistance and screened using the RFP reporter.
 +
 +
 +
 +
 +
 +
 +
 +
 +
 +
----
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 23:51, 7 October 2011

Artificial Yeast Chromosome (Ura3 & Amp)

This part serves as a foundational contribution to expand the registry of yeast components. It is an easy to use artificial yeast chromosome with an auxitrophic uracil selection method for yeast expression, as well as an ampicillin selection for propagation in E.coli. The part was modified for biobrick compatibility from a working pRS316 plasmid containing a Gal1/10 bidirectional promoter which was removed through restriction digest. In addition to the biobrick prefix and suffix, the backbone contains Kpn1 and Sac1 restriction sites the flank the prefix and suffix respectively.

Yeast transformants can be selected by using a uracil drop out medium and E.coli transformants may be selected using ampicillin resistance.



Vgem2011-yeast-transformants.jpg
Vgem2011-ecoli-transformants.jpg



S. cerevisiae (left) Colonies shown are successful yeast transformants using our backbone and BBa_J04450 (which cannot be be expressed as assembled in yeast).  Colonies were selected using a medium lacking uracil.  


E. coli (right) Red transformants indicate successful assembly of the backbone with the registry part BBa_J04450, which contains an RFP reporter protein.  The bacterial colonies were selected using ampicillin resistance and screened using the RFP reporter.





Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal NheI site found at 175
    Illegal NheI site found at 3149
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BglII site found at 4850
    Illegal XhoI site found at 22
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal NgoMIV site found at 7
    Illegal NgoMIV site found at 4858
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.