Table 1.

List of the primers used for vector construction and strain verification. USER-specific overhangs are marked in bold, translational enhancer (Kozak) sequence is underlined (Cavener & Ray, 1991; Nakagawa et al., 2008)

Name	Sequence	Application
pESC_U_ccdB_fw	5′-AAAAGAGCTCGAATGCGTGCGATCGCAG-3′	Amplification of USER cassette, ccdB gene, and chloramphenicol cassette
pESC_U_ccdB_rv	5′-AAAACTCGAGGAATGCACGCGATCGCTG-3′
ID399USERrev	5′-ATTGGGUGCATAGGCCACTAGTGGATCTG-3′	Amplification of LoxP-flanked selection marker cassettes
ID400USERfwd	5′-ATCGCGUCAGCTGAAGCTTCGTACGC-3′
ID401pIntFwdU	5′-ACCCAAUTCGCCCTATAGTGAGTCG-3′	Amplification of integrative plasmid backbone
ID402pintRevU	5′-ACGCGAUCTTCGAGCGTCCCAAAACC-3′
ID1493	5′- CGTGCGAUCCGCATAGGGAGTGTAAATTTATC -3′	Amplification of positive GFP control fragment for USER plasmid verification
ID1494	5′-CACGCGAUAGTGAAAGGAAGGCCCATGAG -3′
PTEF1_fw	5′-ACCTGCACUTTGTAATTAAAACTTAG-3′	Amplification of TEF1 promoter
PTEF1_rv	5′-CACGCGAUGCACACACCATAGCTTC-3′
YFP/CFP_F+	5′-AGTGCAGGUAAAACAATGAGTAAAGGAGAAGAACTTTTCAC-3′	Amplification of YFP and CFP genes
YFP/CFP_R+	5′-CGTGCGAUTCATTTGTATAGTTCATCCATGCCATG-3′
RFP_F+	5′-AGTGCAGGUAAAACAATGGCCTCCTCCGAGGACGTCATC-3′	Amplification of RFP gene
RFP_R+	5′-CGTGCGAUTCAGGCGCCGGTGGAGTGGCGG-3′
ID901 X-2-up-out	5′-TGCGACAGAAGAAAGGGAAG-3′	PCR with ID339 verifies insertion in X-2-UP
ID902-X-2-down-out	5′-GAGAACGAGAGGACCCAACAT-3′	PCR with ID401 verifies insertion in X-2-DW
ID903-X-3-up-out	5′-TGACGAATCGTTAGGCACAG-3′	PCR with ID339 verifies insertion in X-3-UP
ID904-X-3-down-out	5′-CCGTGCAATACCAAAATCG-3′	PCR with ID401 verifies insertion in X-3-DW
ID905-X-4-up-out	5′-CTCACAAAGGGACGAATCCT-3′	PCR with ID339 verifies insertion in X-4-UP
ID906-X-4-down-out	5′-GACGGTACGTTGACCAGAG-3′	PCR with ID401 verifies insertion in X-4-DW
ID339-TEF1_test_rv	5′-GCTCATTAGAAAGAAAGCATAGC-3′	Verification of insertion of constructs containing TEF1