Table 1 Oligonucleotide sequences. Oligonucleotide primers used in expression plasmid construction. The unique order number, short name, DNA sequence, and purpose is shown. Sense EPR, myc, and mycEPR oligonucleotides were separately paired with antisense myc/EPR oligonucleotide for amplification and restriction with XhoI and XbaI. HSA sense and antisense clEPRHV3 were paired for amplification and restriction with SnaBI and XbaI. Additional details are provided in the Materials and Methods
Number | Name | Sequence | Purpose |
|---|---|---|---|
123,620,623 | Sense EPR | 5’-TCTCTCGAG AAAAGAGAGC CTAGAATCAC CTACACAGAC TGC-3’ | Mutate and amplify codons to alter HV3HSA cDNA to EPR-HV3HSA, provide XhoI site |
123,620,624 | Sense myc | 5’-TCTCTCGAGA AAAGAGAACA AAAACTCATC TCAGAAGAGG ATCTGATCAC CTACACAGAC TGC-3′ | Mutate and amplify codons to alter HV3HSA cDNA to myc-HV3HSA, provide XhoI site |
123,620,625 | Sense mycEPR | 5’-TCT CTC GAG AAA AGA GAA CAA AAA CTC ATC TCA GAA GAG GAT CTG GAG CCT AGA ATC ACC TAC ACA GAC TGC-3′ | Mutate and amplify codons to alter HV3HSA cDNA to myc-EPRHV3HSA, provide XhoI site |
ML-08-6225 | Antisense myc/EPR | 5′- CTCTCCAAGC TGCTCGAAAA GCTC-3’ | Anchor 3′ end of amplification in all 3 cases above, allow inclusion of XbaI site in amplicon |
890,140,002 | HSA sense | 5’-AGGCATCCTGA TTACTCTGTC GTGCTGCTG-3’ | Anchor 5′ end of amplification for HSAEPR-HV3 construct, allow inclusion of XbaI site in amplicon |
161,722,273 | Antisense clEPRHV3 | 5’ TGTGTACGTAA TTCTAGGCTC GGATCCTAAG GCAGC-3’ | Mutate and amplify codons specifying EPR cleavage site for HSAEPR-HV3 construct, allow inclusion of SnaBI site in amplicon |