Table 1. Primers used in this study.
| Primer | Use | Sequence |
|---|---|---|
| Subcloning TRIM5α constructs (human, human-v1rhesus, rhesus) into pQCXIP | ||
| oJT029 | Amplify human or rhesus TRIM5α-HA (Fwd), add NotI site, for cloning into pQCXIP | caagcggccgcgccaccATGGCTTCTGGAATC |
| oJT030 | Amplify human or rhesus TRIM5α-HA (Rev), add EcoRI site, for cloning into pQCXIP | gcggaattcTCAagcgtagtctgggacgtc |
| DMS library construction | ||
| oJT037 | Flanking primer for all DMS library construction (Fwd), amplifies pQCXIP backbone 5' of NotI site for Gibson cloning with pQCXIP-TRIM5α digested with NotI and BamHI | acctgcaggaattgatccgcggcc |
| oJT038 | Flanking primer for rhesus DMS library construction (Rev), amplifies rhesus TRIM5α 3' of BamHI site for Gibson cloning with pQCXIP-TRIM5α digested with NotI and BamHI | GGATTGGAAGCCAGCACATACCCCCAG |
| oJT003 | Randomize rhesus TRIM5α at codon Q332 (Fwd primer, use w/oJT038 for C-term half) | CGGAACCCACAGATAATGTATNNSGCACCAGGGACATTATTTAC |
| oJT004 | Randomize rhesus TRIM5α at codon Q332 (Rev primer, use w/oJT037 for N-term half) | GTAAATAATGTCCCTGGTGCSNNATACATTATCTGTGGGTTCCG |
| oJT005 | Randomize rhesus TRIM5α at codon A333 (Fwd primer, use w/oJT038 for C-term half) | GAACCCACAGATAATGTATCAGNNSCCAGGGACATTATTTACGTTTC |
| oJT006 | Randomize rhesus TRIM5α at codon A333 (Rev primer, use w/oJT037 for N-term half) | GAAACGTAAATAATGTCCCTGGSNNCTGATACATTATCTGTGGGTTC |
| oJT007 | Randomize rhesus TRIM5α at codon P334 (Fwd primer, use w/oJT038 for C-term half) | CCACAGATAATGTATCAGGCANNSGGGACATTATTTACGTTTCCG |
| oJT008 | Randomize rhesus TRIM5α at codon P334 (Rev primer, use w/oJT037 for N-term half) | CGGAAACGTAAATAATGTCCCSNNTGCCTGATACATTATCTGTGG |
| oJT009 | Randomize rhesus TRIM5α at codon G335 (Fwd primer, use w/oJT038 for C-term half) | CAGATAATGTATCAGGCACCANNSACATTATTTACGTTTCCGTCAC |
| oJT010 | Randomize rhesus TRIM5α at codon G335 (Rev primer, use w/oJT037 for N-term half) | GTGACGGAAACGTAAATAATGTSNNTGGTGCCTGATACATTATCTG |
| oJT011 | Randomize rhesus TRIM5α at codon T336 (Fwd primer, use w/oJT038 for C-term half) | ATGTATCAGGCACCAGGGNNSTTATTTACGTTTCCGTCACTCAC |
| oJT012 | Randomize rhesus TRIM5α at codon T336 (Rev primer, use w/oJT037 for N-term half) | GTGAGTGACGGAAACGTAAATAASNNCCCTGGTGCCTGATACAT |
| oJT013 | Randomize rhesus TRIM5α at codon L337 (Fwd primer, use w/oJT038 for C-term half) | TATCAGGCACCAGGGACANNSTTTACGTTTCCGTCACTCAC |
| oJT014 | Randomize rhesus TRIM5α at codon L337 (Rev primer, use w/oJT037 for N-term half) | GTGAGTGACGGAAACGTAAASNNTGTCCCTGGTGCCTGATA |
| oJT015 | Randomize rhesus TRIM5α at codon F338 (Fwd primer, use w/oJT038 for C-term half) | CAGGCACCAGGGACATTANNSACGTTTCCGTCACTCACG |
| oJT016 | Randomize rhesus TRIM5α at codon F338 (Rev primer, use w/oJT037 for N-term half) | CGTGAGTGACGGAAACGTSNNTAATGTCCCTGGTGCCTG |
| oJT017 | Randomize rhesus TRIM5α at codon T339 (Fwd primer, use w/oJT038 for C-term half) | AGGCACCAGGGACATTATTTNNSTTTCCGTCACTCACGAATTTC |
| oJT018 | Randomize rhesus TRIM5α at codon T339 (Rev primer, use w/oJT037 for N-term half) | GAAATTCGTGAGTGACGGAAASNNAAATAATGTCCCTGGTGCCT |
| oJT019 | Randomize rhesus TRIM5α at codon F340 (Fwd primer, use w/oJT038 for C-term half) | CACCAGGGACATTATTTACGNNSCCGTCACTCACGAATTTCAAT |
| oJT020 | Randomize rhesus TRIM5α at codon F340 (Rev primer, use w/oJT037 for N-term half) | ATTGAAATTCGTGAGTGACGGSNNCGTAAATAATGTCCCTGGTG |
| oJT021 | Randomize rhesus TRIM5α at codon P341 (Fwd primer, use w/oJT038 for C-term half) | ACCAGGGACATTATTTACGTTTNNSTCACTCACGAATTTCAATTATTGTA |
| oJT022 | Randomize rhesus TRIM5α at codon P341 (Rev primer, use w/oJT037 for N-term half) | TACAATAATTGAAATTCGTGAGTGASNNAAACGTAAATAATGTCCCTGGT |
| oJT023 | Randomize rhesus TRIM5α at codon S342 (Fwd primer, use w/oJT038 for C-term half) | GGGACATTATTTACGTTTCCGNNSCTCACGAATTTCAATTATTGTACT |
| oJT024 | Randomize rhesus TRIM5α at codon S342 (Rev primer, use w/oJT037 for N-term half) | AGTACAATAATTGAAATTCGTGAGSNNCGGAAACGTAAATAATGTCCC |
| oJT025 | Randomize rhesus TRIM5α at codon L343 (Fwd primer, use w/oJT038 for C-term half) | GACATTATTTACGTTTCCGTCANNSACGAATTTCAATTATTGTACTGGC |
| oJT026 | Randomize rhesus TRIM5α at codon L343 (Rev primer, use w/oJT037 for N-term half) | GCCAGTACAATAATTGAAATTCGTSNNTGACGGAAACGTAAATAATGTC |
| oJT027 | Randomize rhesus TRIM5α at codon T344 (Fwd primer, use w/oJT038 for C-term half) | CATTATTTACGTTTCCGTCACTCNNSAATTTCAATTATTGTACTGGCGTC |
| oJT028 | Randomize rhesus TRIM5α at codon T344 (Rev primer, use w/oJT037 for N-term half) | GACGCCAGTACAATAATTGAAATTSNNGAGTGACGGAAACGTAAATAATG |
| oAS024 | Flanking primer for human DMS libraries (Rev), amplifies human TRIM5α 3' of BamHI site for Gibson cloning with pQCXIP-TRIM5α digested with NotI and BamHI | AGCACATACCCCCAGGATCCAAGCAG |
| oAS002 | Amplify N-term half of human TRIM5α (WT or R332P) before codon G330, to hybridize w/randomized G330 C-term half (Rev primer, use w/oJT037) | ATATATTATCTGTGGTTTCGGAGAGC |
| oAS001 | Randomize human TRIM5α (WT) at codon G330 (Fwd primer, use w/oAS024 for C-term half) | GCTCTCCGAAACCACAGATAATATATNNSGCACGAGGGACAAGATACC |
| oJT142 | Randomize human TRIM5α (R332P) at codon G330 (Fwd primer, use w/oAS024 for C-term half) | GCTCTCCGAAACCACAGATAATATATnnsGCACcAGGGACAAGATACCA |
| oAS004 | Amplify N-term half of human TRIM5α (WT or R332P) before codon A331, to hybridize w/randomized A331 C-term half (Rev primer, use w/oJT037) | CCCATATATTATCTGTGGTTTCGG |
| oAS003 | Randomize human TRIM5α (WT) at codon A331 (Fwd primer, use w/oAS024 for C-term half) | CCGAAACCACAGATAATATATGGGNNSCGAGGGACAAGATACCAGA |
| oJT143 | Randomize human TRIM5α (R332P) at codon A331 (Fwd primer, use w/oAS024 for C-term half) | CCGAAACCACAGATAATATATGGGnnsCcAGGGACAAGATACCAGAC |
| oAS006 | Amplify N-term half of human TRIM5α (WT) before codon R332 to hybridizew/randomized R332 C-term half (Rev primer, use w/oJT037) | TGCCCCATATATTATCTGTGGTTTC |
| oAS005 | Randomize human TRIM5α (WT) at codon R332 (Fwd primer, use w/oAS024 for C-term half) | GAAACCACAGATAATATATGGGGCANNSGGGACAAGATACCAGACATTTG |
| oAS008 | Amplify N-term half of human TRIM5α (WT) before codon G333 to hybridizew/randomized G333 C-term half (Rev primer, use w/oJT037) | TCGTGCCCCATATATTATCTGTG |
| oAS007 | Randomize human TRIM5α (WT) at codon G333 (Fwd primer, use w/oAS024 for C-term half) | CACAGATAATATATGGGGCACGANNSACAAGATACCAGACATTTGTGAATT |
| oJT144 | Amplify N-term half of human TRIM5α (R332P) before codon G333 to hybridize w/randomized G333 C-term half (Rev primer, use w/oJT037) | TgGTGCCCCATATATTATCTGTG |
| oJT145 | Randomize human TRIM5α (R332P) at codon G333 (Fwd primer, use w/oAS024 for C-term half) | CACAGATAATATATGGGGCACcAnnsACAAGATACCAGACATTTGTGAATTTC |
| oAS010 | Amplify N-term half of human TRIM5α (WT) before codon T334 to hybridizew/randomized T334 C-term half (Rev primer, use w/oJT037) | CCCTCGTGCCCCATATATTA |
| oAS009 | Randomize human TRIM5α (WT) at codon T334 (Fwd primer, use w/oAS024 for C-term half) | TAATATATGGGGCACGAGGGNNSAGATACCAGACATTTGTGAATTTCA |
| oJT146 | Amplify N-term half of human TRIM5α (R332P) before codon T334 to hybridizew/randomized T334 C-term half (Rev primer, use w/oJT037) | CCCTgGTGCCCCATATATTATCTG |
| oJT147 | Randomize human TRIM5α (R332P) at codon T334 (Fwd primer, use w/oAS024 for C-term half) | CAGATAATATATGGGGCACcAGGGnnsAGATACCAGACATTTGTGAATTTCAATTA |
| oAS012 | Amplify N-term half of human TRIM5α (WT) before codon R335 to hybridizew/randomized R335 C-term half (Rev primer, use w/oJT037) | TGTCCCTCGTGCCCCAT |
| oAS011 | Randomize human TRIM5α (WT) at codon R335 (Fwd primer, use w/oAS024 for C-term half) | ATggggcacgagggacaNNStaccagacatttgtgAATTTCAATTATTG |
| oJT148 | Amplify N-term half of human TRIM5α (R332P) before codon R335 to hybridizew/randomized R335 C-term half (Rev primer, use w/oJT037) | TGTCCCTgGTGCCCCATATA |
| oJT149 | Randomize human TRIM5α (R332P) at codon R335 (Fwd primer, use w/oAS024 for C-term half) | TATATGGGGCACcAGGGACAnnsTACCAGACATTTGTGAATTTCAATTATTG |
| oAS014 | Amplify N-term half of human TRIM5α (WT) before codon Y336 to hybridizew/randomized Y336 C-term half (Rev primer, use w/oJT037) | TCTTGTCCCTCGTGCCC |
| oAS013 | Randomize human TRIM5α (WT) at codon Y336 (Fwd primer, use w/oAS024 for C-term half) | GGGCACGAGGGACAAGANNSCAGACATTTGTGAATTTCAATTATTG |
| oJT150 | Amplify N-term half of human TRIM5α (R332P) before codon Y336 to hybridize w/randomized Y336 C-term half (Rev primer, use w/oJT037) | TCTTGTCCCTgGTGCCCC |
| oJT151 | Randomize human TRIM5α (R332P) at codon Y336 (Fwd primer, use w/oAS024 for C-term half) | GGGGCACcAGGGACAAGAnnsCAGACATTTGTGAATTTCAATTATTGTAC |
| oAS016 | Amplify N-term half of human TRIM5α (WT) before codon Q337 to hybridize w/randomized Q337 C-term half (Rev primer, use w/oJT037) | GTATCTTGTCCCTCGTGCC |
| oAS015 | Randomize human TRIM5α (WT) at codon Q337 (Fwd primer, use w/oAS024 for C-term half) | GCACGAGGGACAAGATACNNSACATTTGTGAATTTCAATTATTGTACTG |
| oJT152 | Amplify N-term half of human TRIM5α (R332P) before codon Q337 to hybridize w/randomized Q337 C-term half (Rev primer, use w/oJT037) | GTATCTTGTCCCTgGTGCC |
| oJT153 | Randomize human TRIM5α (R332P) at codon Q337 (Fwd primer, use w/oAS024 for C-term half) | GGCACcAGGGACAAGATACnnsACATTTGTGAATTTCAATTATTGTACTGG |
| oAS018 | Amplify N-term half of human TRIM5α (WT) before codon T338 to hybridize w/randomized T338 C-term half (Rev primer, use w/oJT037) | CTGGTATCTTGTCCCTCGTG |
| oAS017 | Randomize human TRIM5α (WT) at codon T338 (Fwd primer, use w/oAS024 for C-term half) | CACGAGGGACAAGATACCAGNNSTTTGTGAATTTCAATTATTGTACTGG |
| oJT154 | Amplify N-term half of human TRIM5α (R332P) before codon T338 to hybridize w/randomized T338 C-term half (Rev primer, use w/oJT037) | CTGGTATCTTGTCCCTgGTG |
| oJT155 | Randomize human TRIM5α (R332P) at codon T338 (Fwd primer, use w/oAS024 for C-term half) | CACcAGGGACAAGATACCAGnnsTTTGTGAATTTCAATTATTGTACTGGCAT |
| oAS020 | Amplify N-term half of human TRIM5α (WT) before codon F339 to hybridizew/randomized F339 C-term half (Rev primer, use w/oJT037) | TGTCTGGTATCTTGTCCCTC |
| oAS019 | Randomize human TRIM5α (WT) at codon F339 (Fwd primer, use w/oAS024 for C-term half) | GAGGGACAAGATACCAGACANNSGTGAATTTCAATTATTGTACTGGC |
| oJT156 | Amplify N-term half of human TRIM5α (R332P) before codon F339 to hybridizew/randomized F339 C-term half (Rev primer, use w/oJT037) | TGTCTGGTATCTTGTCCCTgG |
| oJT157 | Randomize human TRIM5α (R332P) at codon F339 (Fwd primer, use w/oAS024 for C-term half) | CcAGGGACAAGATACCAGACAnnsGTGAATTTCAATTATTGTACTGGCATC |
| oAS022 | Amplify N-term half of human TRIM5α (WT) before codon V340 to hybridizew/randomized V340 C-term half (Rev primer, use w/oJT037) | AAATGTCTGGTATCTTGTCCCTC |
| oAS021 | Randomize human TRIM5α (WT) at codon V340 (Fwd primer, use w/oAS024 for C-term half) | AGGGACAAGATACCAGACATTTNNSAATTTCAATTATTGTACTGGCATCC |
| oJT158 | Amplify N-term half of human TRIM5α (R332P) before codon V340 to hybridizew/randomized V340 C-term half (Rev primer, use w/oJT037) | AAATGTCTGGTATCTTGTCCCTg |
| oJT159 | Randomize human TRIM5α (R332P) at codon V340 (Fwd primer, use w/oAS024 for C-term half) | cAGGGACAAGATACCAGACATTTnnsAATTTCAATTATTGTACTGGCATCCTG |
| Illumina library construction | ||
| oJT055 | Illumina library construction, PCR1 Fwd primer (rhesus TRIM5α only), amplifies v1 loop and adds adaptor | tcgtcggcagcgtcagatgtgtataagagacagTGAGCTCTCGGAACCCACAGATAATGTAT |
| oJT056 | Illumina library construction, PCR1 Rev primer (rhesus TRIM5α only), amplifies v1 loop and adds adaptor | gtctcgtgggctcggagatgtgtataagagacagGCCCAGGACGCCAGTACAATAATTGAAATT |
| oJT113 | Illumina library construction, PCR1 Fwd primer (human TRIM5α libraries) amplifies v1 loop and adds adaptor | tcgtcggcagcgtcagatgtgtataagagacagCAAGTGAGCTCTCCGAAACCACAGATAATATAT |
| oJT114 | Illumina library construction, PCR1 Rev primer (human TRIM5α libraries), amplifies v1 loop and adds adaptor | gtctcgtgggctcggagatgtgtataagagacagGAGCCCAGGATGCCAGTACAATAATTGAAATT |
| oJT057 | Illumina library construction, PCR2 Fwd primer (all libraries), adds P5 adaptor | AATGATACGGCGACCACCGAGATCTACACtagatcgcTCGTCGGCAGCGTC |
| oJT058 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N701 barcoode | CAAGCAGAAGACGGCATACGAGATtcgccttaGTCTCGTGGGCTCGG |
| oJT059 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N702 barcoode | CAAGCAGAAGACGGCATACGAGATctagtacgGTCTCGTGGGCTCGG |
| oJT060 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N703 barcoode | CAAGCAGAAGACGGCATACGAGATttctgcctGTCTCGTGGGCTCGG |
| oJT115 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N704 barcoode | CAAGCAGAAGACGGCATACGAGATgctcaggaGTCTCGTGGGCTCGG |
| oJT116 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N705 barcoode | CAAGCAGAAGACGGCATACGAGATaggagtccGTCTCGTGGGCTCGG |
| oJT117 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N706 barcoode | CAAGCAGAAGACGGCATACGAGATcatgcctaGTCTCGTGGGCTCGG |
| oJT118 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N707 barcoode | CAAGCAGAAGACGGCATACGAGATgtagagagGTCTCGTGGGCTCGG |
| oJT119 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N708 barcoode | CAAGCAGAAGACGGCATACGAGATcctctctgGTCTCGTGGGCTCGG |
| oJT138 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N709 barcoode | CAAGCAGAAGACGGCATACGAGATagcgtagcGTCTCGTGGGCTCGG |
| oJT139 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N710 barcoode | CAAGCAGAAGACGGCATACGAGATcagcctcgGTCTCGTGGGCTCGG |
| oJT140 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N711 barcoode | CAAGCAGAAGACGGCATACGAGATtgcctcttGTCTCGTGGGCTCGG |
| oJT141 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N712 barcoode | CAAGCAGAAGACGGCATACGAGATtcctctacGTCTCGTGGGCTCGG |
| oJT166 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N714 barcoode | CAAGCAGAAGACGGCATACGAGATTCATGAGCGTCTCGTGGGCTCGG |
| oJT167 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N715 barcoode | CAAGCAGAAGACGGCATACGAGATCCTGAGATGTCTCGTGGGCTCGG |
| oJT168 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N716 barcoode | CAAGCAGAAGACGGCATACGAGATTAGCGAGTGTCTCGTGGGCTCGG |
| oJT169 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N718 barcoode | CAAGCAGAAGACGGCATACGAGATGTAGCTCCGTCTCGTGGGCTCGG |
| oJT170 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N719 barcoode | CAAGCAGAAGACGGCATACGAGATTACTACGCGTCTCGTGGGCTCGG |
| oJT171 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N720 barcoode | CAAGCAGAAGACGGCATACGAGATAGGCTCCGGTCTCGTGGGCTCGG |
| oJT172 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N721 barcoode | AAGCAGAAGACGGCATACGAGATGCAGCGTAGTCTCGTGGGCTCGG |
| oJT173 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N722 barcoode | CAAGCAGAAGACGGCATACGAGATCTGCGCATGTCTCGTGGGCTCGG |
| oJT174 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N723 barcoode | CAAGCAGAAGACGGCATACGAGATGAGCGCTAGTCTCGTGGGCTCGG |
| oJT175 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N724 barcoode | CAAGCAGAAGACGGCATACGAGATCGCTCAGTGTCTCGTGGGCTCGG |
| oJT176 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N726 barcoode | CAAGCAGAAGACGGCATACGAGATGTCTTAGGGTCTCGTGGGCTCGG |
| oJT177 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N727 barcoode | CAAGCAGAAGACGGCATACGAGATACTGATCGGTCTCGTGGGCTCGG |
| oJT178 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N728 barcoode | CAAGCAGAAGACGGCATACGAGATTAGCTGCAGTCTCGTGGGCTCGG |
| oJT179 | Illumina library construction, PCR2 Rev primer (all libraries), adds P7 adaptor and N729 barcoode | CAAGCAGAAGACGGCATACGAGATGACGTCGAGTCTCGTGGGCTCGG |
| RhT5-IlluminaF | Custom sequencing primer for rhesus TRIM5α Illumina libraries, sequences rhesus TRIM5α v1 loop (39 nt) | TGAGCTCTCGGAACCCACAGATAATGTAT |
| HsT5-IlluminaF | Custom sequencing primer for human TRIM5α Illumina libraries, sequences human TRIM5α v1 loop (33 nt) | CAAGTGAGCTCTCCGAAACCACAGATAATATAT |
| Quikchange PCR for targeted mutagenesis | ||
| oCY001 | Generate G330E mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY002) | CTCCGAAACCACAGATAATATATGaGGCACGAGGGACAAGATAC |
| oCY002 | Generate G330E mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY001) | GTATCTTGTCCCTCGTGCCtCATATATTATCTGTGGTTTCGGAG |
| oCY003 | Generate A331E mutation in human TRIM5α F(wd primer, amplify full plasmid with oCY004) | GAAACCACAGATAATATATGGGGaACGAGGGACAAGATACCAG |
| oCY004 | Generate A331E mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY003) | CTGGTATCTTGTCCCTCGTtCCCCATATATTATCTGTGGTTTC |
| oCY005 | Generate R332E mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY006) | ACCACAGATAATATATGGGGCAgaAGGGACAAGATACCAGACATT |
| oCY006 | Generate R332E mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY005) | AATGTCTGGTATCTTGTCCCTtcTGCCCCATATATTATCTGTGGT |
| oJT040 | Generate R332P mutation in human TRIM5α (Fwd primer, amplify full plasmid with oJT041) | CCACAGATAATATATGGGGCACcAGGGACAAGATACCAGACATTTG |
| oJT041 | Generate R332P mutation in human TRIM5α (Rev primer, amplify full plasmid with oJT040) | CAAATGTCTGGTATCTTGTCCCTgGTGCCCCATATATTATCTGTGG |
| oCY007 | Generate G333Y mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY008) | CACAGATAATATATGGGGCACGAtacACAAGATACCAGACATTTGTGAATT |
| oCY008 | Generate G333Y mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY007) | AATTCACAAATGTCTGGTATCTTGTgtaTCGTGCCCCATATATTATCTGTG |
| oCY009 | Generate G333D mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY010) | CAGATAATATATGGGGCACGAGatACAAGATACCAGACATTTGTGAATTTC |
| oCY010 | Generate G333D mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY009) | GAAATTCACAAATGTCTGGTATCTTGTatCTCGTGCCCCATATATTATCTG |
| oCY011 | Generate T334D mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY012) | GATAATATATGGGGCACGAGGGgacAGATACCAGACATTTGTGAATTTC |
| oCY012 | Generate T334D mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY011) | GAAATTCACAAATGTCTGGTATCTgtcCCCTCGTGCCCCATATATTATC |
| oJT246 | Generate R335A mutation in human TRIM5α (Fwd primer, amplify full plasmid with oJT247) | TATGGGGCACGAGGGACAgcATACCAGACATTTGTGAATTTCAATTATTG |
| oJT247 | Generate R335A mutation in human TRIM5α (Rev primer, amplify full plasmid with oJT246) | CAATAATTGAAATTCACAAATGTCTGGTATgcTGTCCCTCGTGCCCCATA |
| oCY013 | Generate R335E mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY014) | TGGGGCACGAGGGACAgaATACCAGACATTTGTGAATTTCAATTATTG |
| oCY014 | Generate R335E mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY013) | CAATAATTGAAATTCACAAATGTCTGGTATtcTGTCCCTCGTGCCCCA |
| oCY015 | Generate Y336E mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY016) | GGGCACGAGGGACAAGAgAaCAGACATTTGTGAATTTCAATTATTGTAC |
| oCY016 | Generate Y336E mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY015) | GTACAATAATTGAAATTCACAAATGTCTGtTcTCTTGTCCCTCGTGCCC |
| oCY017 | Generate Q337D mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY018) | GGCACGAGGGACAAGATACgAtACATTTGTGAATTTCAATTATTGTACTG |
| oCY018 | Generate Q337D mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY017) | CAGTACAATAATTGAAATTCACAAATGTaTcGTATCTTGTCCCTCGTGCC |
| oAS2019-05 | Generate Q337N mutation in human TRIM5α (Fwd primer, amplify full plasmid with oAS2019-06) | GGCACGAGGGACAAGATACaacACATTTGTGAATTTCAATTATTGTACTGG |
| oAS2019-06 | Generate Q337N mutation in human TRIM5α (Rev primer, amplify full plasmid with oAS2019-05) | CCAGTACAATAATTGAAATTCACAAATGTgttGTATCTTGTCCCTCGTGCC |
| oCY019 | Generate T338E mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY020) | ACGAGGGACAAGATACCAGgaATTTGTGAATTTCAATTATTGTACTGGC |
| oCY020 | Generate T338E mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY019) | GCCAGTACAATAATTGAAATTCACAAATtcCTGGTATCTTGTCCCTCGT |
| oCY021 | Generate F339E mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY022) | GAGGGACAAGATACCAGACAgaaGTGAATTTCAATTATTGTACTGGCA |
| oCY022 | Generate F339E mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY021) | TGCCAGTACAATAATTGAAATTCACttcTGTCTGGTATCTTGTCCCTC |
| oCY023 | Generate V340E mutation in human TRIM5α (Fwd primer, amplify full plasmid with oCY024) | GGGACAAGATACCAGACATTTGaGAATTTCAATTATTGTACTGGCATCC |
| oCY024 | Generate V340E mutation in human TRIM5α (Rev primer, amplify full plasmid with oCY023) | GGATGCCAGTACAATAATTGAAATTCtCAAATGTCTGGTATCTTGTCCC |
| oAS2019-07 | Generate V340H mutation in human TRIM5α (Fwd primer, amplify full plasmid with oAS2019-08) | GAGGGACAAGATACCAGACATTTcacAATTTCAATTATTGTACTGGCATCCT |
| oAS2019-08 | Generate V340H mutation in human TRIM5α (Rev primer, amplify full plasmid with oAS2019-07) | AGGATGCCAGTACAATAATTGAAATTgtgAAATGTCTGGTATCTTGTCCCTC |
| oJT162 | Generate V340stop mutation in human TRIM5α (Fwd primer, amplify full plasmid with oJT163) | GAGGGACAAGATACCAGACATTTtaGAATTTCAATTATTGTACTGGCATCC |
| oJT163 | Generate V340stop mutation in human TRIM5α (Rev primer, amplify full plasmid with oJT162) | GGATGCCAGTACAATAATTGAAATTCtaAAATGTCTGGTATCTTGTCCCTC |
| oJT248 | Generate R335A double mutation in human TRIM5α-R332P (Fwd primer, amplify full plasmid with oJT249) | TATGGGGCACcAGGGACAgcATACCAGACATTTGTGAATTTCAATTATTG |
| oJT249 | Generate R335A double mutation in human TRIM5α-R332P (Rev primer, amplify full plasmid with oJT248) | CAATAATTGAAATTCACAAATGTCTGGTATgcTGTCCCTgGTGCCCCATA |
| oAS2019-03 | Generate R335E double mutation in human TRIM5α-R332P (Fwd primer, amplify full plasmid with oAS2019-04) | TGGGGCACcAGGGACAgaaTACCAGACATTTGTGAATTTCAATTATTG |
| oAS2019-04 | Generate R335E double mutation in human TRIM5α-R332P (Rev primer, amplify full plasmid with oAS2019-03) | CAATAATTGAAATTCACAAATGTCTGGTAttcTGTCCCTgGTGCCCCA |
| oAS2019-01 | Generate Q337D double mutation in human TRIM5α-R332P (Fwd primer, amplify full plasmid with oAS2019-02) | GGCACcAGGGACAAGATACgacACATTTGTGAATTTCAATTATTGTACTGG |
| oAS2019-02 | Generate Q337D double mutation in human TRIM5α-R332P (Rev primer, amplify full plasmid with oAS2019-01) | CCAGTACAATAATTGAAATTCACAAATGTgtcGTATCTTGTCCCTgGTGCC |
| oAS2019-09 | Generate Q337N double mutation in human TRIM5α-R332P (Fwd primer, amplify full plasmid with oAS2019-10) | GGCACcAGGGACAAGATACaacACATTTGTGAATTTCAATTATTGTACTGG |
| oAS2019-10 | Generate Q337N double mutation in human TRIM5α-R332P (Rev primer, amplify full plasmid with oAS2019-09) | CCAGTACAATAATTGAAATTCACAAATGTgttGTATCTTGTCCCTgGTGCC |
| oJT122 | Generate Q332E mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT123) | CTCGGAACCCACAGATAATGTATgAGGCACCAGGGACATTATTTAC |
| oJT123 | Generate Q332E mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT122) | GTAAATAATGTCCCTGGTGCCTcATACATTATCTGTGGGTTCCGAG |
| oJT120 | Generate A333E mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT121) | GAACCCACAGATAATGTATCAGGaACCAGGGACATTATTTACGTTTCC |
| oJT121 | Generate A333E mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT120) | GGAAACGTAAATAATGTCCCTGGTtCCTGATACATTATCTGTGGGTTC |
| oCY063 | Generate A333W mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oCY064) | GAACCCACAGATAATGTATCAGtggCCAGGGACATTATTTACGTTTC |
| oCY064 | Generate A333W mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oCY063) | GAAACGTAAATAATGTCCCTGGCCACTGATACATTATCTGTGGGTTC |
| oJT124 | Generate P334M mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT125) | CCCACAGATAATGTATCAGGCAatgGGGACATTATTTACGTTTCCGTC |
| oJT125 | Generate P334M mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT124) | GACGGAAACGTAAATAATGTCCCcatTGCCTGATACATTATCTGTGGG |
| oJT095 | Generate G335I mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT096) | ACAGATAATGTATCAGGCACCAatcACATTATTTACGTTTCCGTCACTC |
| oJT096 | Generate G335I mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT095) | GAGTGACGGAAACGTAAATAATGTgatTGGTGCCTGATACATTATCTGT |
| oJT097 | Generate T336Q mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT098) | GATAATGTATCAGGCACCAGGGcaaTTATTTACGTTTCCGTCACTCAC |
| oJT098 | Generate T336Q mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT097) | GTGAGTGACGGAAACGTAAATAAttgCCCTGGTGCCTGATACATTATC |
| oJT099 | Generate L337N mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT100) | GTATCAGGCACCAGGGACAaacTTTACGTTTCCGTCACTCACG |
| oJT100 | Generate L337N mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT99) | CGTGAGTGACGGAAACGTAAAgttTGTCCCTGGTGCCTGATAC |
| oJT103 | Generate F338K mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT104) | TCAGGCACCAGGGACATTAaagACGTTTCCGTCACTCACG |
| oJT104 | Generate F338K mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT103) | CGTGAGTGACGGAAACGTcttTAATGTCCCTGGTGCCTGA |
| oJT126 | Generate F338Q mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT127) | TCAGGCACCAGGGACATTAcagACGTTTCCGTCACTCACGA |
| oJT127 | Generate F338Q mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT126) | TCGTGAGTGACGGAAACGTctgTAATGTCCCTGGTGCCTGA |
| oJT105 | Generate T339F mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT106) | CAGGCACCAGGGACATTATTTttcTTTCCGTCACTCACGAATTTCA |
| oJT106 | Generate T339F mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT105) | TGAAATTCGTGAGTGACGGAAAgaaAAATAATGTCCCTGGTGCCTG |
| oJT128 | Generate T339Q mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT129) | CAGGCACCAGGGACATTATTTcaGTTTCCGTCACTCACGAATTTC |
| oJT129 | Generate T339Q mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT128) | GAAATTCGTGAGTGACGGAAACtgAAATAATGTCCCTGGTGCCTG |
| oJT130 | Generate F340D mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT131) | GCACCAGGGACATTATTTACGgaTCCGTCACTCACGAATTTCAATTA |
| oJT131 | Generate F340D mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT130) | TAATTGAAATTCGTGAGTGACGGAtcCGTAAATAATGTCCCTGGTGC |
| oJT132 | Generate P341G mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT133) | CACCAGGGACATTATTTACGTTTggGTCACTCACGAATTTCAATTATTGT |
| oJT133 | Generate P341G mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT132) | ACAATAATTGAAATTCGTGAGTGACccAAACGTAAATAATGTCCCTGGTG |
| oJT107 | Generate P341I mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT108) | CACCAGGGACATTATTTACGTTTataTCACTCACGAATTTCAATTATTGTAC |
| oJT108 | Generate P341I mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT107) | GTACAATAATTGAAATTCGTGAGTGAtatAAACGTAAATAATGTCCCTGGTG |
| oJT109 | Generate P341IK mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT110) | CACCAGGGACATTATTTACGTTTaaaTCACTCACGAATTTCAATTATTGTAC |
| oJT110 | Generate P341IK mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT109) | GTACAATAATTGAAATTCGTGAGTGAtttAAACGTAAATAATGTCCCTGGTG |
| oJT134 | Generate S342G mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT135) | AGGGACATTATTTACGTTTCCGggACTCACGAATTTCAATTATTGTACTG |
| oJT135 | Generate S342G mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT134) | CAGTACAATAATTGAAATTCGTGAGTccCGGAAACGTAAATAATGTCCCT |
| oJT101 | Generate T344E mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT102) | CATTATTTACGTTTCCGTCACTCgagAATTTCAATTATTGTACTGGCGTC |
| oJT102 | Generate T344E mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT101) | GACGCCAGTACAATAATTGAAATTctcGAGTGACGGAAACGTAAATAATG |
| oJT111 | Generate T344stop mutation in rhesus TRIM5α (Fwd primer, amplify full plasmid with oJT112) | CATTATTTACGTTTCCGTCACTCtagAATTTCAATTATTGTACTGGCGTC |
| oJT112 | Generate T344stop mutation in rhesus TRIM5α (Rev primer, amplify full plasmid with oJT111) | GACGCCAGTACAATAATTGAAATTctaGAGTGACGGAAACGTAAATAATG |
| Sequencing primers | ||
| pQCXIP-F | Sequencing primer from pQCXIP backbone 5' of multiple cloning site (Fwd) | acaccgggaccgatccag |
| HsT5-midF | Sequencing primer from midpoint of human TRIM5α (Fwd) | GATCTGGAGCATCGGCTG |
| HsT5-midR | Sequencing primer from midpoint of human TRIM5α (Rev) | CAAGGTCACGTTCTCCGTC |
| RhT5-midF | Sequencing primer from midpoint of rhesus TRIM5α (Fwd) | CTCATCTCAGAACTGGAGCATC |
| RhT5-midR | Sequencing primer from midpoint of rhesus TRIM5α (Rev) | CTTCAAGGTCATGTTCTCAATCC |