Part:BBa_K2171001:Design
RvLEAM - 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
"RvLEAM CDS + transcription terminators" subpart was ordered as gBlock from IDT
Sequence: ATATAATTGAATTCGCGGCCGCTTCTAGATGTTTTTAGCTCGTAATGCTGGCCGTGCAGGCTA TCGCGGTGTAGTGGCGTATCAGCAAGCCGCTTCTTTCAGCGTGAGTTCCGCAAAAGCTGCCGG ATCACGCTCGTCTGGGGGGAGTGACGCGGGAGACTATGCCCGCGAGGCCGCGGAGCACGCTAA GGCAGGATTGAAGGACCTTAAAAACGAGGCGTCTTGGAAAGCTAAAGGAGTTGCGAACCAAGC CGCCGGTGCTTTTGAGCGTGCTAAAGATACTGTGAAGGAGGGGGTCCACGACATGAAACGCTC AGGTAGTCGCGTTTTCGAACAAGGACAAGAGGAAGTGGAAGCCGGTGCTCAGCACGCTAAGGC AGGCTACCAAAGCGCCAAAAATGCGGCACAGGATACGGCTGCTACATTGAAGGACAAGGCGGG GTCCGCTTGGAACCAGGCCAAACACGTTGTGGAAGACAAGGGTGAGGACGTTGTCGAAGCTGT TAAAGACACCGCAAGTAAGGTCTGGGGAAAAGCCAAACACGTTGCTGAGGACGTAAAGGAGAA TGCGCAATCCCCCGGCGGAATCGCTGACAAAGCCTCGGATGTATGGTCCGCAGCTAAGGATAA AGCTGCGGACGTGCTGAGCGGCGCTAAGCACACGGCCGAGAACTTGGCGCACAAGGCGCAAGC CGCTATCCACGATGCAACGGCATCTAGCGGGTCCCAGTCACAGTCTCAATCCCAATCACAATA TCGCCAGGGCCAACAGCAAGGTCGTCAAGATCAACAACAAAGCAAAAGCCAATGGGGACAGAC CTCACCCCAGTCGCCTGACGGGTTTCGTCCACAAGCTGGTCAAGGTCCCCAGGGCGGGAAAGG GCCGGGACAGGCCGGAGGCCGTCGTTAATAATTGATTAACATCAAATAAAACGAAAGGCTCAG TCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCTACTAGAGTCAC ACTGGCTCACCTTCGGGTGGGCCTTTCTGCGTTTATTACTAGTAGCGGCCGCTGCAGTTATAT AT
and amplified using primers:
F: gagtcacacaggaaagtactagATGTTTTTAGCTCGTAATGCTGG
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.