Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Bakul Jitendra Vinchhi, Sarah Veugelen, Katrien Vandermeeren, Yana Hoorne
Date created: 2011-09-15 11:00:00
Date modified: 2015-05-08 01:12:47
OmpA_transmembrane_domain + Linker + point_mutant_HPLC6(AFP)
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K584019_sequence (Version 1) |
Description
This composite part contains the two sequences OmpA and Point Mutant HPLC6(Antifreeze Protein - AFP) that are linked via a flexible linker.AFP normally is expressed intracellularly to prevent freezing of the cytoplasm, but in our system, it was essential that AFP was extracellularly anchored at the cell membrane, thereby preventing ice formation outside the cell. Therefore, we merged the AFP gene to OmpA via a flexible linker, thereby ensuring the desired extracellular localization of AFP.
Notes
A point mutation at Base Pair number 156 of the HPLC6 sequence was performed by replacing the T in the wild-type coding sequence to C.This mutation was necessary as the sequence CTGCAG in the Wild-type coding sequence corresponded to a Pst1 restriction site. GCT(Wild type) and GCC(1 point mutant) are both valid triplet codons for Alanine.Source
SOURCE - Pseudopleuronectes americanus (winter flounder)Gene - HPLC6
| Sequence Annotation | Location | Component / Role(s) |
| OmpA Flexible_Linker HPLC6(AFP) HPLC6_Point_Mutant | 1,434 435,459 460,708 615,615 | feature/protein CDS sequence_feature feature/misc feature/protein CDS sequence_alteration feature/mutation |