TABLE 1.

Oligonucleotides used to create p220 lox-scanning mutants

p220 mutantPrimer descriptionPrimer sequence (5′-3′)a
1 (LoxP121-145)1st round, forwardGCC CGA ACG AGA ACT GGA ATT GCA GAA ATC AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseTGT ACC TGT GGA AGG AGG AGT GGT AAA CTG TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardTCC CCA AGG TTT GCT GGC AGT CAG AGA GCC CGA ACG AGA ACT GGA ATT GCA GAA ATC
2nd round, reverseAAT TTG GCC ACT TGG TCG AGT AAC CTG TGT ACC TGT GGA AGG AGG AGT GGT AAA CTG
2 (LoxP225-249)1st round, forwardTCT CAG AGA AAA AGT ACC ACT TTG TCT GGC AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseTTG AAG AGA TTT GTT GCT TAG TAT TTT TTC TCG TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardTCT CCC GGT AGA CGC AAA AGT GAA TCT CAG AGA AAA AGT ACC ACT TTG TCT GGC
2nd round, reverseTTT ATT TAT GTT TTC TGC TAG CTT TTC TTG AAG AGA TTT GTT GCT TAG TAT TTT TTC TCG
3 (LoxP325-349)1st round, forwardTTG GAA CAG ACA GAA TCA GAC CCA GCA TTT AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseGAC TAT ACT GGG ATT GGA TTC CAT AGG TTG TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardATG TCT GAA GAA GCT ATA CAG GAC ATA TTG GAA CAG ACA GAA TCA GAC CCA GCA TTT
2nd round, reverseAAC TGC TAG ATT AGT TTC ATC TGC TAA GAC TAT ACT GGG ATT GGA TTC CAT AGG TTG
4 (LoxP422-446)1st round, forwardGAA GAC CAG GAA AAT TTT TCC CAA ATA AGT ACC AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseCCC TCT TTG GTT AAA GTC ATT CAA ATT AGG CAC TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardAGC AAC AAC CAT GAT GTG CTT AGA CAA GAA GAC CAG GAA AAT TTT TCC CAA ATA AGT ACC
2nd round, reverseATG TGG ATT ACA TTC AGC ATT AGA ATT CCC TCT TTG GTT AAA GTC ATT CAA ATT AGG CAC
5 (LoxP629-653)1st round, forwardGGA CAA GTA GAA ATT CAT CTT GGA GAT TCG AGG GAA CAA AAG CTG GGT ACC ATA A TCG
1st round, reverseAGG CTC CTG TGA ATT CTC AGA CTT GGA TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardGTT GAA AGT TCA CAT TTA AAT GTA TCT GGA CAA GTA GAA ATT CAT CTT GGA GAT TCG
2nd round, reverseATT CTC TTC TTT TAC AGA AGA TGA AGG CTC CTG TGA ATT CTC AGA CTT GGA
6 (LoxP727-752)1st round, forwardTCC CAA AAT ACT GAT GAT AAA CCT TCT AGC AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseACT AGA AAC AGC ACT GGT AAG TTC AGT ATC TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardTCT TCA GTG GGA GAT TCT CAC CCT GAG TCC CAA AAT ACT GAT GAT AAA CCT TCT AGC
2nd round, reverseTAT AGT TGG CAG GTT TTC TCC ATT AAT ACT AGA AAC AGC ACT GGT AAT TTC AGT ATC
7 (LoxP828-853)1st round, forwardTCT GAA GAC TCT GCA GTA AAC AAT ACT CAG AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseTGA ATT GCC AAA AGC TGT GCT TGT GGC TGG TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardATG GAA CAG AGT CTT TTA ACA TTC AAA TCT GAA GAC TCT GCA GTA AAC AAT ACT CAG
2nd round, reverseCAC ACA GGT AGC TAT CAG AAT GTT ATT TGA ATT GCC AAA AGC TGT GCT TGT GGC TGG
8 (LoxP926-950)1st round, forwardTTT GCT GTC AAC CAA GCT GTG TCA CCA AAC AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseGTT ATT TCC ATT CTG TCC AAC CAC AGA TAC TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardCAG ACA CCA CCA AGG TCA AAC AGT GTA TTT GCT GTC AAC CAA GCT GTG TCA CCA AAC
2nd round, reverseAAG AAC CTG CCG AGG AGG AGT AGA AAA GTT ATT TCC ATT CTG TCC AAC CAC AGA TAC
9 (LoxP1039-1054)1st round, forwardTGT CAT GCA CAA AAA ACT GAA GTT TCT GAC AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseTAC ACG TCT GTG GCA TGG TTT AGC AGC TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardGTG GAT TCG TCA GGT CAT TCA GTT GGA TGT CAT GCA CAA AAA ACT GAA GTT TCT GAC
2nd round, reverseCAC AGG AGC AGT AGT GCT GTC GAA ACA GAG TAC ACG TCT GTG GCA TGG TTT AGC AGC
10 (LoxP1125-1149)1st round, forwardATC AAA AGA GAG AAA GAG AAG CCT CCT CTG AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseAGA TTC AAG CAC AAT TTC TGT TGG TGA AAC TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardACC TTA AAA CCC CCT TCT AAT AAT GCT ATC AAA AGA GAG AAA GAG AAG CCT CCT CTG
2nd round, reverseCTC CTT ATT AGC TGT TGC TTT ATG GAA AGA TTC AAG CAC AAT TTC TGT TGG TGA AAC
11 (LoxP1228-1252)1st round, forwardAAA AAT GTA CTT TCA GTA GGT ACA GCT GTG AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseACT ATC AGC AAG CCT ACT TAC TGA GCT GTG TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardAAA AAA CAA GGC ACA TCT TCA AAC AAT AAA AAT GTA CTT TCA GTA GGT ACA GCT GTG
2nd round, reverseAGG TGT CCG GGG CAC AGG TAA ATC ACT ACT ATC AGC AAG CCT ACT TAC TGA GCT GTG
12 (LoxP1325-1349)1st round, forwardAGC CCT GCC AGT GAA ACA GGA AGT AGG GAA CAA AAG CTG GGT ACC ATA ACT TCG
1st round, reverseAAA CTG TTG TGT GTT ATC TTT CAG AGG AGT TAT CGA ATT CCC GGG ATC CGA TAA CTT CG
2nd round, forwardCCT GTC ACC CCA GAC TTG CCT GCC TGC AGC CCT GCC AGT GAA ACA GGA AGT
2nd round, reverseTTT TGT GGT GCT CCT TGA AGA TGC TCT AAA CTG TTG TGT GTT ATC TTT CAG AGG AGT
  • a Nucleotides in italics correspond to sequences homologous to the Tcr gene used for selection.