Difference between revisions of "Part:BBa K3582500"
 
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This version of Pet28a(+) vector is stringently modified by removing all the unnecessary tags and non required regions. The insertion site is aptly annotated with a start and a stop codon where the insertion of the ORF can be made. Most importantly RFC10 criteria are satisfied and all of the other features of the vector are conserved and undisturbed.
 
This version of Pet28a(+) vector is stringently modified by removing all the unnecessary tags and non required regions. The insertion site is aptly annotated with a start and a stop codon where the insertion of the ORF can be made. Most importantly RFC10 criteria are satisfied and all of the other features of the vector are conserved and undisturbed.
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This part can be widely and extensively used for protein expression efficiently without complexities.
  
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3582500 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3582500 SequenceAndFeatures</partinfo>
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===References===
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*https://plasmid.med.harvard.edu/PLASMID/GetVectorDetail.do?vectorid=319
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*http://dnasu.org/DNASU/GetVectorDetail.do?vectorid=319
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Latest revision as of 17:36, 24 October 2020


Modified Pet28a(+) expression vector

pET28a(+) is a novel expression plasmid generally used in E-coli strains for protein production purposes. It has a kanamycin resistance gene as a selection marker. Plasmid sequence consists of the T7 promoter and terminator sequences as well as lac operon sequence and lac promoter for protein expression induction. Apart from this, it also has well-defined operator, promoter, MCS and tag sequences precisely inbuilt.

The normal pet28a(+) vector contains several unnecessary components like 6Xhis tag, thrombin site etc. that could interfere with protein expression and purification. Identifying a proper insertion site becomes difficult. The normal version does not even satisfy the RFC10 criteria.

This version of Pet28a(+) vector is stringently modified by removing all the unnecessary tags and non required regions. The insertion site is aptly annotated with a start and a stop codon where the insertion of the ORF can be made. Most importantly RFC10 criteria are satisfied and all of the other features of the vector are conserved and undisturbed. This part can be widely and extensively used for protein expression efficiently without complexities.



Figure 1. Visualization of modified Pet28a(+) vector with complete annotation.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 105
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 137
    Illegal NgoMIV site found at 1725
    Illegal NgoMIV site found at 1885
    Illegal NgoMIV site found at 4932
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 2805


References