Part:BBa_K2171002:Design
MASH - osmotic stress protein from tardigrade
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
The "lacI-lacIq and a tac promoter" subpart was amplified from part BBa_K731520 using primers:
F: tatgaattcgcggccgcttctagagTCACTGCCCGCTTTCCAGTC
R: CTAGTACTTTCCTGTGTGACTC
"MAHS CDS + transcription terminators" subpart was ordered as gBlock from IDT
Sequence: ATATAATTGAATTCGCGGCCGCTTCTAGATGGTTTCGTTGCGTTTAGGTGATTTCCGCTCGCAGCCAAGTTTACGTAGCGTGCCAGC GAGCTTAACTTCCCGCAGTCAGGCGTTCAGCCTTCAGGAGATTGCAGCGCGCGCGGGGGTAGTACTGCGTGGCGTGCAACAACAGTT CCGTAACGTAACGGGAGTGAATGCTGCCCCGGTGGTGGCCTTTGACAACGGCTCAGTCCTTTATTCTGAGCGCATCCATAGCCAATC ATCTCAAAAGCAGGCCCCCACAACAGTCCCCACAGGCAGCGTCAGCAATAGTCCTCAACCGGAGGGGAAAGCCAACGAAGCCGCCGA GCGTGCCAAACAATTTATGAACCCACCCGTAGCCCCAATGGACCCAGTCGACAAAAATGAGTTCGTGGCTATGCCCGAGATGGGGCG CTCCAACGGAAATGGCGAGAACAAGCAGGCAGCGGACTTTATGAAGAATCAGGGCGATACTGACATGGACTCACAGTACGCCCCAGA CTCCAGTAAGAACACGAAGTCTGTCCCGACGAAGGAAATCGTGGCAGAGGACGGAAGTATGTCTATTGAGGATATTAAAAAAGCGAC GCAGGTTACTCCAGGGGTCGCGGTTAAAAATGAGGGGGTATAATAATTGATTAACATCAAATAAAACGAAAGGCTCAGTCGAAAGAC TGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCTACTAGAGTCACACTGGCTCACCTTCGGGTGGGCCTTTCTGCGTT TATTACTAGTAGCGGCCGCTGCAGTTATATAT
and amplified using primers:
F: gagtcacacaggaaagtactagATGGTTTCGTTGCGTTTAGGTG
R: taactgcagCGGCCGCTACTAGTAATAAACGC
Both subparts were fused together by Overlap Extention PCR. All PCRs were performed with Phusion High Fidelity Polymerase using the standart protocol provided by the manufacturer. The final part was gel purified.The final part was gel purified and cloned in standart vector via restriction (EcoRI and PstI) and ligation.
Note: Uppercase characters in primer sequences correspond to regions that are complementary to the intended matrix. Lowercase characters mark regions of overlap for OEP and/or parts of biobrick regions with restriction sites for EcoRI and PstI.