[http://partsregistry.org/Part:BBa_929107 BBa_929107]
Anti-GFP, IgG1 Fc with TEV site, TMD, mCherry, framed by LoxP
Antibody Module
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This part was derived by de novo synthesis based on sequence information provided by GenBank and UniProt and by one already existing Biobrick part. The antibody construct consists of two major building blocks represented by the actual antibody unit (BBa_K929102) and the switchable membrane anchoring region (BBa_K929103). Both elements guarantee the eligibility and the easy handling of the construct and are optimized for expression in CHO cells. The antibody unit is represented by the human Ig kappa chain V-I region signal peptide (UniProt: P01601), the anti-GFP Nanobody (PDB: 3OGO) and the Fc region (UniProt: P01857). TEV protease recognition site, 2 LoxP sites, the B-cell receptor transmembrane domain (modified BBa_K157010) and the mCherry reporter display the switchable membrane anchoring region. The TEV recognition site on protein level and the LoxP sites on the genetically level allow the shift from surface presentation to secretion of the antibody unit. For detailed information see the [http://partsregistry.org/Part:BBa_K929003 partsregistry], for experimental experience also see All Biobricks.
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[http://partsregistry.org/Part:BBa_K929003 BBa_K929003]
modified AID with CMV, hGH-polyA and eGFP
Mutation Module
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This part is an improved version of wildtype AID, the enzyme that randomly mutates predominantly in the immunoglobulin genes. It is designed for strong expression of the fusion protein modified AID+eGFP. Modified AID has an additional Nuclear Localization Sequence (NLS)and the naturally occurring Nuclear Export Sequence (NES)is deleted. Due to the fact that AID mutates the actively transcribed single stranded DNA, it is supposed that the direction of the enzyme to the inside of the nucleus would improve the mutation rate.The fusion with the green fluorescent reporter eGFP allows 1.)to check the transfection success 2.)select transfected cells via FACS and 3.)to check the cellular localization of the fusion protein. For detailed information see the [http://partsregistry.org/Part:BBa_K929003 partsregistry], for experimental experience also see All Biobricks.
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[http://partsregistry.org/Part:BBa_K929301 BBa_K929301]
Potsdam Standard Backbone
Potsdam Standard
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This part is an improved version of wildtype AID, the enzyme that randomly mutates predominantly in the immunoglobulin genes. It is designed for strong expression of the fusion protein modified AID+eGFP. Modified AID has an additional Nuclear Localization Sequence (NLS)and the naturally occurring Nuclear Export Sequence (NES)is deleted. Due to the fact that AID mutates the actively transcribed single stranded DNA, it is supposed that the direction of the enzyme to the inside of the nucleus would improve the mutation rate.The fusion with the green fluorescent reporter eGFP allows 1.)to check the transfection success 2.)select transfected cells via FACS and 3.)to check the cellular localization of the fusion protein. For detailed information see the [http://partsregistry.org/Part:BBa_K929003 partsregistry], for experimental experience also see All Biobricks.
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