Coding

Part:BBa_K2171003:Design

Designed by: Slavil Peykov, Desislava Popova   Group: iGEM16_Bulgaria   (2016-10-14)


SAHS2 - osmotic stress protein from tardigrade with periplasmic secretion signal peptide


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


To create this part you can follow the guidelines for part BBa K2171004 (described here: https://parts.igem.org/Part:BBa_K2171004:Design) with some modifications.


Modifications:


1. Use gBlock with sequence: 

ATATAATTGAATTCGCGGCCGCTTCTAGATGAAAAAACTGCTGTTCGCGATT
CCGCTGGTGGTGCCGTTCTATAGCCATAGCGCAGATGACGCAGCTCACGAGG
AAGGGGTGGAGTGGACGGGTAAGCCGTGGATGGGAAAATGGGAGTCAGATCC
TTCTAAGGATGAGAATGTAGAAGAGTTTAAGAAAAAATTGCAGTTACCTATG
TCCCATTCGGAAATGAACAAGAACTCCAAAGTCTGGATTCACCACTATAAAA
AAGGCGACGAATACCATCATAAGATTATTATCAATGACGCGCATTACAAAAA
TGATATCGTGTTCAAATTGGGACAAGAGAGTGCCGGGTCGTATAATGGCTCA
TCATTCTCCGTGAAATACGAAGACAAAGACGGTGCTTTAGTTGGAAGCGTAC
ACTACACGGGTACAAAAGAGCAAAGTCTTGATAAAACCATCAATAATGTATT
CAAGCTGGAGGGCGACCACCTGGTGAAGACGTCCACCATTGAGGGGGTTACC
ATGAAGCGCCACTACAACAAACGTCAGTAATAATTGATTAACATCAAATAAA
ACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCG
GTGAACGCTCTCTACTAGAGTCACACTGGCTCACCTTCGGGTGGGCCTTTCT
GCGTTTATTACTAGTAGCGGCCGCTGCAGTTATATAT


2. Use new F primer for the gBlock amplification:

New F: gagtcacacaggaaagtactagATGAAAAAACTGCTGTTCGCG


Note: Uppercase characters in primer sequence corresponds to regions that are complementary to the intended matrix. Lowercase characters mark region of overlap for OEP.