Generator

Part:BBa_K880005

Designed by: Josh Atkinson, Mike Ferguson, and Ben Parker   Group: iGEM12_Michigan   (2012-09-29)

Strong promoter, strong RBS combination for high expression levels of proteins

The part is composed of sub-parts J23100 and B0034.

This is a strong promoter, strong RBS combination for high expression levels of proteins. It has a consensus constitutive promoter and RBS sequence-produces strongest possible expression.

Use of this Part by Team Michigan '12

Team Michigan '12 used this part as their primary protein-generator. Our data suggests that we were successful in using it to express FimE and HBiF recombinases for our recombinase-based switch systems. See our results page for more information[1].

Use of this Part by Team UBrawijaya '21

CHOP

UBrawijaya iGEM 2021 project CHOP systemis an outer membrane vesicle based protein overexpression system that includes three goals towards producing a blueprint for an ideal protein production system. This project can be customized because the system can be adapted with various proteins of interest, the overexpression system that can increase protein yield, and the clean harvest with simplified purification steps that is equipped with a protease enzyme expression mechanism in the concentrated system - from vesicles and cell pellets.

This part is used to tune the expression level of constitutively expressed parts. We decided to characterize two different primary protein-generators to our system; Constitutive promoter family member-RBS and T7 promoter-RBS.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Functional Parameters



Functional Parameters: Austin_UTexas

Burden Imposed by this Part:

Burden Value: 27.5 ± 8.6%

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This part exhibited a significant burden. Users should be aware that BioBricks with a burden of >20-30% may be susceptible to mutating to become less functional or nonfunctional as an evolutionary consequence of this fitness cost. This risk increases as they used for more bacterial cell divisions or in larger cultures. Users should be especially careful when combining multiple burdensome parts, as plasmids with a total burden of >40% are expected to mutate so quickly that they become unclonable. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.

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