Table 2 List of primers used in this study
From: New inducible promoter for gene expression and synthetic biology in Yarrowia lipolytica
Gene/names | Primers | Sequences | |
|---|---|---|---|
P1-EYK | EYK-P-F | GTTGTGTGATGAGACCTTGGTGC | Deletion of EYK and verification of EYK deletion |
P2-EYK | EYK-P-R-SfiI | AAAGGCCATTTAGGCCGCAGCTCCTCCGACAATCTTG | |
T1-EYK | EYK-T-L-SfiI | TAAGGCCTTGATGGCCACAAGTAGAGGGAGGAGAAGC | |
T2-EYK | EYK-T-R | GTTTAGGTGCCTGAAGACGGTG | |
LPR-L-SfiI | ATAGGCCTAAATGGCCTGCATCGATCTAGGGATAACAGG | ||
LPR-R-SfiI | ATAGGCCATCAAGGCCGCTAGATAGAGTCGAGAATTACCCTG | ||
EYK-V1 | EYK-V1 | CGTACCCGAGATTGTACTGTTGTC | |
EYK-V2 | EYK-V2 | CATAACCGCCTACCCTTGTAGC | |
P300 | pEYK300 F | GACATCGATGCATCTACTTTTCTCTATACTGT | |
P300 | pEYK R | GACGGATCCAGTAGATGTGTAAGTGTGTAGAAG | Promoter ampli, ClaI |
P450 | MT-TATAampli-F | ACGATCGATTTTGTGCAAGTGTGTGTGTGTG | Promoter ampli, BamHI |
P450 | MT-TATAampli-R | ACGACTAGTCAGGTCATCGGATTATGCAAGG | Promoter ampli, ClaI |
ST043 | pEYK-mut1 | CGATGCATCTACTTTTCTCTATACTGTACGTTTCAATCTGGGGAAGCGGAATCCCAAAAGGGAAAGCCGCCGCATTAAGCTCCACAGCC | Promoter ampli, SpeI |
ST044 | pEYK-mut1BIS | CGATGCATCTACTTTTCTCTATACTGTACGTTTCAATCTGGGGAAGCGGAATCCCAAAAGG ACGCGT CGCCGCATTAAGCTCCACAGCC | WT bloc 1 (domain A) |
ST045 | pEYK-mut2 | TTGCATAATCCGATGACCTGA | Mutated bloc 1(domain a) |
ST046 | pEYK-mut2BIS | TTG TACGCGTA GATGACCTGA | WT bloc 2 (domain B) |
ST047 | pEYK-mutA | GGCGTAATTCGAGGTGTCGGAACGTATTAGGCTACTGGACTGATC | Mutated bloc 2 (domain b) |
ST048 | pEYK-mutABIS | GGCGTAATTCGAGGTGTCGGAAC ATGCGCAT CTACTGGACTGATC | WT blocA- complementary strand (domain B) |
ST049 | pEYK-mutB | TACGTAGATGAAAAGAGATATGACATGCAAAGTTAGACCCCTTCGCCTTAGGGTTTTCCCTTTCGCG | Mutated bloc A- complementary strand (domain b) |
ST050 | pEYK-mutBbis | TACGTAGATGAAAAGAGATATGACATGCAAAGTTAGACCCCTTCGCCTTAGGGTTTTCC TGCGCA CG | WT bloc B- complementary strand (domain A) |
MT-URA3-for | GCGTAGGTGAAGTCGTCAAT | Mutated bloc B- complementary strand (domain a) | |
MT-YFP-rev | CAGATGAACTTCAGGGTCAGC | For promoter sequence verification (forward) | |
pTEF-start | GGGTATAAAAGACCACCGTCC | For promoter sequence verification (reverse) | |
61stop | GTAGATAGTTGAGGTAGAAGTTG | For gene verification (forward) | |
For gene verification (reverse) |