Difference between revisions of "Part:BBa K3037002"

 
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<partinfo>BBa_K3037001 short</partinfo>
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
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! colspan="2" style="background:#FFBF00;"|Maltose Binding Protein
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|-
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|'''Function'''
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|Expression
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|-
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|'''Use in'''
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|Escherichia coli
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|-
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|'''RFC standard'''
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|RFC 25 compatible
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|-
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|'''Backbone'''
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|pSB1C3<br>
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|-
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|'''Submitted by'''
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|Team:TU_Dresden 2019[https://2019.igem.org/Team:TU_Dresden]
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|}
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=== Overview ===
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The TU Dresden 2019 team design this biobrick in order to make a fusion protein with dCas9 in accordance to the RFC 25 standard. [https://2019.igem.org/Team:TU_Dresden (more information)]
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MBP was inserted into the pSB1C3 vector for transformation and expressed in E. coli .
  
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__NOTOC__
 
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<partinfo>BBa_K3037002 short</partinfo>
 
  
 
There are many dCas9 biobricks already available but all of them are optimized for expression in mammalian cells. This is the first one that is codon optimized to be expressed in e.coli.  
 
There are many dCas9 biobricks already available but all of them are optimized for expression in mammalian cells. This is the first one that is codon optimized to be expressed in e.coli.  
 
New scope of in vitro applications of cas9, which is normally used in vivo mainly
 
New scope of in vitro applications of cas9, which is normally used in vivo mainly
  
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=== Biology ===
  
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<!-- Add more about the biology of this part here
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Part of the CRISPR System
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===Usage and Biology===
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Immune system of bacteria, which store sequences of viral infections, recognize them and cut them apart
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Mutation to not cut
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We can use the system by providing guideRNAs to locate to any target
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Implemented in many engineering approaches of mammalian cells
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But we make it possible to be used by overexpressing it in microbes
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Can bind any sequence of interest given by the guide RNA
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Has mutation in the Ruv site and therefore no endonuclease function (only binding no cutting)
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=== Sequence ===
  
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<!-- -->
 
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<span class='h3bb'>Sequence and Features</span>
 
 
<partinfo>BBa_K3037002 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3037002 SequenceAndFeatures</partinfo>
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=== Design Notes ===
  
  
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<!-- Uncomment this to enable Functional Parameter display
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=== References ===
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===Functional Parameters===
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<partinfo>BBa_K3037002 parameters</partinfo>
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<!-- -->
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Revision as of 16:24, 3 October 2019

Maltose Binding Protein (MBP-tag)

Maltose Binding Protein
Function Expression
Use in Escherichia coli
RFC standard RFC 25 compatible
Backbone pSB1C3
Submitted by Team:TU_Dresden 2019[1]


Overview

The TU Dresden 2019 team design this biobrick in order to make a fusion protein with dCas9 in accordance to the RFC 25 standard. (more information)

MBP was inserted into the pSB1C3 vector for transformation and expressed in E. coli .


There are many dCas9 biobricks already available but all of them are optimized for expression in mammalian cells. This is the first one that is codon optimized to be expressed in e.coli. New scope of in vitro applications of cas9, which is normally used in vivo mainly

Biology

Part of the CRISPR System Immune system of bacteria, which store sequences of viral infections, recognize them and cut them apart Mutation to not cut We can use the system by providing guideRNAs to locate to any target Implemented in many engineering approaches of mammalian cells But we make it possible to be used by overexpressing it in microbes Can bind any sequence of interest given by the guide RNA Has mutation in the Ruv site and therefore no endonuclease function (only binding no cutting)


Sequence


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1096
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3375
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Design Notes

References