Table 1.

Primers used in the construction of the p2KO ablation/insertion vector

Primer name	Sequence
left flank
66R L SacI (f)	5′-AATGGATCACATAAAGGAGCTCTTAACG-3′
66R L NotI (r)	5′- CAGAAAACATGCGGCCGCATATAATCT-3′
right flank
66R R EcoRI	5′-GGAGATGAACAAGAAATAGAATTCATAGG-3′
66R R HindIII	5′- CTGTTCTTTATCACAAGCTTCTATCGGGTG-3′
mGM-CSF
hmGMCSF BamHI (f)	5′-TAGGCCTGGGATCCGATCCACCGGTCGCCACCATGTGGCTGCAGA-3′
mGMCSF XmaI (r)	5′-CTCATCAATGTATCTTATCATCCCGGGCTAGCT-3′
mCCL2/MCP-1
mMCP-1 BamHI (f)	5′-TAGGCCTGGGATCCGATCCACCGGTCGCCACCATGCAGGTCCCTG-3′
mMCP-1 XmaI (r)	5′-CGGCGATCCCCGGGAGATACTAGTTCAC-3′
fliC (S. typhimurium)
FliC BamHI (f)	5′-ACCCGGGGATCCTCTAGAAATAATTTTG-3′
FliC XmaI (r)	5′-GGAGCTCGAACCCGGGTCCTTAAC-3′
M13 (f)	5′-TGTAAAACGACGGCCAGT-3′
M13 (r)	5′-CAGGAAACAGCTATGACC-3′
66R ablation verification
66R int (f)	5′- CGGTATCAAATTGCTAGGTATACTTGC-3′
66R int (r)	5′- CTCCAATTCGTTTAGAAAACGATGCG-3′
2L ablation verification
2L int (f)	5′- CCATTGCATCCTTCAGAACAAG-3′
2L int (r)	5′- GCATAACTTTAAAATATAATTATACTGTTACG-3′
PCR template control
136R int (f)	5′- GTATTTATGTACTGTTTCAACTAACAAAAGC-3′
136R int (r)	5′- CCTTTAGGTGTTAGGATATATCAATTATACAG-3′

Where applicable the inserted restriction endonucleases sites are indicated by italicized text and start codons within the forward primer and/or the stop codons within the reverse primer are shown by gray shading. The 66R int (f/r) and 2L int (f/r) primer sets are verification primers used to verify 66R and/or 2L ablations, while the 136R gene is present in all viruses and is used as a positive control for the quality of the template DNA in PCR reactions.