Sequencing Primers
Addgene uses a number of primers for sequence verification of deposited plasmids. Below is a list of primers currently in use. This list is available for your convenience, and will be updated as needed.
Looking for Primers?
The primer sequences listed on the left are provided for your reference. Addgene does not distribute primers.
For sequencing plasmids in our repository, we've chosen primers based on the plasmid backbone and insert. To identify primers that may be useful in your sequencing reaction, find your plasmid page and see what primers are listed under "5' sequencing primer" and "3' sequencing primer".
Still not sure what primer you need? Email us at help@addgene.org
For reference information, please consult Addgene’s Molecular Biology Reference Page
Commonly Used Primers
| CMV Forward | CGCAAATGGGCGGTAGGCGTG (Invitrogen) Human CMV immediate early promoter, forward primer |
| LKO.1 5’ | GACTATCATATGCTTACCGT (Weinberg Lab) Human U6 promoter, forward primer |
| LucNrev | CCTTATGCAGTTGCTCTCC 5’ end of luciferase, reverse primer |
| M13 Reverse | CAGGAAACAGCTATGAC In lacZ gene |
| MSCV | CCCTTGAACCTCCTCGTTCGACC (BD Biosciences) Murine stem cell virus, forward primer |
| pBABE 5’ | CTTTATCCAGCCCTCAC (Weinberg Lab) Psi packaging signal, 5’ of MCS in pBABE vectors, forward primer |
| pGEX 5’ | GGGCTGGCAAGCCACGTTTGGTG 3’ end of glutathione-S-transferase, forward primer |
| SP6 | ATTTAGGTGACACTATAG SP6 promoter, forward primer |
| T3 | GCAATTAACCCTCACTAAAGG T3 promoter, forward primer |
| T7 | TAATACGACTCACTATAGGG T7 promoter, forward primer |
Full Primer List
| 3’AOX1 | GCAAATGGCATTCTGACATCC (Invitrogen) For Pichia vectors with AOX1 terminator, reverse primer |
| 5’AOX1 | GACTGGTTCCAATTGACAAGC (Invitrogen) For Pichia vectors with AOX1 promoter, forward primer |
| 35S promoter | CTATCCTTCGCAAGACCCTTC CaMV 35S promoter, forward primer |
| AC5 | ACACAAAGCCGCTCCATCAG (Invitrogen) Drosophila Actin 5C promoter, forward primer |
| Alpha-factor | TACTATTGCCAGCATTGCTGC (Invitrogen) Alpha factor signal sequence, forward primer |
| Amp-R | ATAATACCGCGCCACATAGC 5’ end of ampicillin resistance gene, reverse primer |
| AUG1 Forward | CAATTTACATCTTTATTTATTAACG (Invitrogen) For Pichia vectors with AUG1 promoter, forward primer |
| AUG1 Reverse | GAAGAGAAAAACATTAGTTGGC (Invitrogen) For Pichia vectors with AUG1 promoter, reverse primer |
| BGH Reverse | TAGAAGGCACAGTCGAGG (Invitrogen) Bovine growth hormone terminator, reverse primer |
| Bglob-intron-F | CTGGTCATCATCCTGCCTTT Rabbit beta-globin intron, forward primer |
| Bglob-intron-R | TTTGCCCCCTCCATATAACA Rabbit beta-globin intron, reverse primer |
| Bglob-pA-R | TTTTGGCAGAGGGAAAAAGA Rabbit beta-globin polyA region, reverse primer |
| CAT-R | GCAACTGACTGAAATGCCTC 5’ end of chloramphenicol resistance gene, reverse primer |
| CMV Forward | CGCAAATGGGCGGTAGGCGTG (Invitrogen) Human CMV immediate early promoter, forward primer |
| CRE-R | GCAAACGGACAGAAGCATTT 5’ end of Cre recombinase, reverse primer |
| CYC1 | GCGTGAATGTAAGCGTGAC (Invitrogen) CYC1 transcription termination signal, reverse primer |
| DsRed1-C | AGCTGGACATCACCTCCCACAACG (BD Biosciences) 3’ end of DsRed1, forward primer |
| DsRed1-N | GTACTGGAACTGGGGGGACAG (BD Biosciences) 5’ end of DsRed1, reverse primer |
| EBV Reverse | GTGGTTTGTCCAAACTCATC (Invitrogen) SV40 polyA terminator, reverse primer |
| Ecdysone Forward | CTCTGAATACTTTCAACAAGTTAC (Invitrogen) Drosophila heat shock promoter, forward primer |
| EF-1a Forward | TCAAGCCTCAGACAGTGGTTC (Invitrogen) Human elongation factor-1a promoter, forward primer |
| EGFP-C | CATGGTCCTGCTGGAGTTCGTG (BD Biosciences) 3’ end of EGFP, forward primer |
| EGFP-N | CGTCGCCGTCCAGCTCGACCAG (BD Biosciences) 5’ end of EGFP, reverse primer |
| EXFP-R | GTCTTGTAGTTGCCGTCGTC (Golenbock lab) For distinguishing EGFP vs ECFP vs EYFP, reverse primer |
| F1ori-F | GTGGACTCTTGTTCCAAACTGG F1 origin, forward primer |
| GAL1 | AATATACCTCTATACTTTAACGTC (Invitrogen) S. cerevisiae GAL1 promoter, forward primer |
| Gal10pro-F | GGTGGTAATGCCATGTAATATG (Stratagene) S. cerevisiae GAL10 promoter, forward primer |
| Gal4 N-term | GAGTAGTAACAAAGGTCAA 3’ end of Gal4 DNA binding domain, forward primer |
| Gal4-AD | AATACCACTACAATGGAT (BD Biosciences) 3’ end of Gal4 activation domain, forward primer |
| GFP-F | GGTCCTTCTTGAGTTTGTAAC 3’ end of GFP, forward primer |
| GFP-R | CCATCTAATTCAACAAGAATTGGGACAAC (Ahmad lab) 5’ end of GFP, reverse primer |
| GPDpro-F | CGGTAGGTATTGATTGTAATTCTG S. cerevisiae GPD promoter, forward primer |
| GW-3’ | GCATGATGACCACCGATATG 3’ end of Gateway cassette, forward primer |
| GW-5’ | AATCTCGCCGGATCCTAACT 5’ end of Gateway cassette, reverse primer |
| H1 | TCGCTATGTGTTCTGGGAAA Human H1 promoter, forward primer |
| HA-F | TACCCATACGACGTCCCAGA HA tag, forward primer |
| HA-R | TCTGGGACGTCGTATGGGTA HA tag, reverse primer |
| HAT | GAGGAGCACGCTCATGCCCAC (BD Biosciences) Histidine affinity tag, forward primer |
| hGH-PA-R | CCAGCTTGGTTCCCAATAGA Human growth hormone terminator, reverse primer |
| hrGFP-R | TCCCCGAGTACCACTTCATC hrGFP (humanized Renilla GFP), forward primer |
| hUBCpro-F | TGAAGCTCCGGTTTTGAACT Human Ubiquitin C (UbC) promoter, forward primer |
| IRES-F | TGGCTCTCCTCAAGCGTATT 3’ end of IRES, forward primer |
| IRES-R | CCTCACATTGCCAAAAGACG 5’ end of IRES, reverse primer |
| L4440 | AGCGAGTCAGTGAGCGAG (Caldwell lab) 5’ of MCS in L4440 vector, forward primer |
| LacI-R | GGCATACTCTGCGACATCGT 5’ end of LacI, reverse primer |
| LacZ-R | GACAGTATCGGCCTCAGGAA 5’ end of LacZ, reverse primer |
| LexA | CGTCAGCAGAGCTTCACCATTG (Caldwell lab) 3’ end of LexA DNA binding domain, forward primer |
| LKO.1 5’ | GACTATCATATGCTTACCGT (Weinberg Lab) Human U6 promoter, forward primer |
| LNCX | AGCTCGTTTAGTGAACCGTCAGATC (BD Biosciences) Human CMV promoter, forward primer |
| Luc-F | AGTCAAGTAACAACCGCGA 3’ end of luciferase, forward primer |
| LucNrev | CCTTATGCAGTTGCTCTCC 5’ end of luciferase, reverse primer |
| M13 (-21) Forward | TGTAAAACGACGGCCAGT In lacZ gene |
| M13 (-40) | GTTTTCCCAGTCACGAC In lacZ gene |
| M13 Reverse | CAGGAAACAGCTATGAC In lacZ gene |
| M13/pUC Forward | CCCAGTCACGACGTTGTAAAACG (Invitrogen) In lacZ gene |
| M13/pUC Reverse | AGCGGATAACAATTTCACACAGG (Invitrogen) In lacZ gene |
| MBP-F | GATGAAGCCCTGAAAGACGCGCAG (Waugh lab) 3’ end of maltose binding protein, forward primer |
| mCherry-F | CCCCGTAATGCAGAAGAAGA 3' end of mCherry, forward primer |
| mCherry-R | TTGGTCACCTTCAGCTTGG 5' end of mCherry, reverse primer |
| MT Forward | CATCTCAGTGCAACTAAA (Invitrogen) Drosophila metallothionein promoter, forward primer |
| MMLV-F | ATCAGTTCGCTTCTCGCTTC Moloney murine leukemia virus LTR (MoMuLV), forward primer |
| mPGK-F | CATTCTGCACGCTTCAAAAG Mouse PGK promoter, forward primer |
| MSCV | CCCTTGAACCTCCTCGTTCGACC (BD Biosciences) Murine stem cell virus, forward primer |
| MSCV-rev | CAGCGGGGCTGCTAAAGCGCATGC Murine stem cell virus, reverse primer |
| MT1-F | GCTGTCCTCTAAGCGTCACC Mouse metallothionein 1 promoter, forward primer |
| mU6-F | CAGCACAAAAGGAAACTCACC Mouse U6 promoter, forward primer |
| Myc | GCATCAATGCAGAAGCTGATCTCA (BD Biosciences) Myc tag, forward primer |
| Neo-F | CGTTGGCTACCCGTGATATT 3’ end of neomycin resistance gene, forward primer |
| Neo-R | GCCCAGTCATAGCCGAATAG 5’ end of neomycin resistance gene, reverse primer |
| NOS-F | GCGTTCAAAAGTCGCCTAAG Nopaline synthase promoter, forward primer |
| Nmt1-F | GCAATGTGCAGCGAAACTAA S. pombe nmt1 promoter, forward primer |
| OpIE2 Forward | CGCAACGATCTGGTAAACAC (Invitrogen) OpIE2 promoter, forward primer |
| pACYC-F | TGAAGTCAGCCCCATACGAT p15A origin, forward primer |
| pAd-CMV | GCTAGAGATCTGGTACCGTC For cloning sites after SalI in pAd-CMV vector |
| pBABE 3’ | ACCCTAACTGACACACATTCC (Weinberg Lab) SV40 enhancer, 3’ of MCS in pBABE vectors, reverse primer |
| pBABE 5’ | CTTTATCCAGCCCTCAC (Weinberg Lab) Psi packaging signal, 5’ of MCS in pBABE vectors, forward primer |
| pBAD Forward | ATGCCATAGCATTTTTATCC (Invitrogen) For vectors with E. coli araBAD promoter, forward primer |
| pBAD Reverse | GATTTAATCTGTATCAGG (Invitrogen) For vectors with E. coli araBAD promoter, reverse primer |
| pBluescriptKS | TCGAGGTCGACGGTATC For pBluescript vector |
| pBluescriptSK | TCTAGAACTAGTGGATC For pBluescript vector |
| pBMN 5’ | GCTTGGATACACGCCGC MMLV sequence, for inserts in pBMN retroviral vector |
| pBR322ori-F | GGGAAACGCCTGGTATCTTT pBRS322 origin, forward primer |
| pBRforBam | CTTGGAGCCACTATCGAC In pBR322 tet region, upstream of BamHI, forward primer |
| pBRforEco | AATAGGCGTATCACGAGGC In pBR322, upsteam of EcoRI site, forward primer |
| pBRrevBam | GGTGATGTCGGCGATATAGG In pBR322 tet region, downstream of BamHI, reverse primer |
| pCAG-F | GCAACGTGCTGGTTATTGTG Rabbit beta-globin intron, for pCAG plasmids, forward primer |
| pCasper-F | GGGTTTTATTAACTTACAT (Vosshall lab) 5’ end of Drosophila mini-white gene, reverse primer |
| pCasper-hs | GCAACTACTGAAATCTGCCAAG Drosophila Hsp70 promoter, forward primer |
| pcDL-F | GTTGCCTTTACTTCTAGGCCT (Kinet lab) 5’ of EcoRI site in pcDL vector, forward primer |
| pENTR-F | CTACAAACTCTTCCTGTTAGTTAG 5’ of attL1 in pENTR vector, forward primer |
| pENTR-R | ATGGCTCATAACACCCCTTG 3’ of attL2 in pENTR vector, reverse primer |
| pGEX 3’ | CCGGGAGCTGCATGTGTCAGAGG 3’ of MCS in pGEX vectors, reverse primer |
| pGEX 5’ | GGGCTGGCAAGCCACGTTTGGTG 3’ end of glutathione-S-transferase, forward primer |
| pGP704-R | AACAAGCCAGGGATGTAACG R6K gamma origin, 3’ of MCS in pGP704 vector, reverse primer |
| pHybLex Reverse | GAGTCACTTTAAAATTTGTATACAC (Invitrogen) ADH terminator, reverse primer |
| pLTet-F | ACTGAGCACATCAGCAGGAC Lambda phage early leftward (pL) promoter, forward primer |
| pLXSN 5’ | CCCTTGAACCTCCTCGTTCGACC (MSCV) Murine stem cell virus, same as MSCV, forward primer |
| pMRB101-F | AAGATGCAGGCAGCTGAGTT HCMV major immediate-early protein (IE), forward primer |
| pMT2-F | TTGCCTTTCTCTCCACAGGT 3’ end of synthetic intron, forward primer |
| pMX-S1811 | GACGGCATCGCAGCTTGGATACAC (Yamanaka lab) MMLV sequence, 5’ of MCS in pMXs vector, forward primer |
| Polyhedrin forward | AAATGATAACCATCTCGC (Invitrogen) Polyhedrin promoter, forward primer |
| Polyhedrin reverse | GTCCAAGTTTCCCTG (Invitrogen) For baculovirus vector with polyhedrin promoter, reverse primer |
| pQE promoter | CCCGAAAAGTGCCACCTG (Qiagen) 5’ of MCS in pQE vectors, forward primer |
| pREP Forward | GCTCGATACAATAAACGCC (Invitrogen) Rous sarcoma virus (RSV) promoter, forward primer |
| pRS-marker | CGGCATCAGAGCAGATTGTA To sequence yeast selectable marker in pRS vectors |
| Pry1 | CTTAGCATGTCCGTGGGGTTTGAAT PZ P-element, reverse primer |
| pTrcHis Forward | GAGGTATATATTAATGTATCG (Invitrogen) 5’ of MCS in pTrcHis vector, forward primer |
| pTrcHis Reverse | GATTTAATCTGTATCAGG (Invitrogen) 3’ of MCS in pTrcHis vector, same as pBAD-R, reverse primer |
| Puro-F | GCAACCTCCCTTCTACGAGC 3’ end of puromycin resistance gene, forward primer |
| pZIP | TCCTTTCCAGCGAGGTTCTA Murine leukemia virus (MuLV), reverse primer |
| RCAS-F | ACATGGGTGGTGGTATAGCGCTTGCG (Orsulic lab) 3’ of Rous sarcoma virus (RSV) env gene, forward primer |
| Rluc-F | CCAGGATTCTTTTCCAATGC 3’ end of Renilla luciferase, forward primer |
| RVprimer3 | CTAGCAAAATAGGCTGTCCC (Promega) 5’ of MCS in pGL3 vector, forward primer |
| SFFV-F | ATTGATTGACTGCCCACCTC Spleen focus forming virus 5’ LTR, forward primer |
| SP6 | ATTTAGGTGACACTATAG SP6 promoter, forward primer |
| SV40pA-R | GAAATTTGTGATGCTATTGC SV40 polyA, reverse primer |
| SV40pro-F | TATTTATGCAGAGGCCGAGG SV40 promoter/origin, forward primer |
| SV40-spliceR | CACAAAGATCCGGACCAAAG SV40 splice sequence, reverse primer |
| T3 | GCAATTAACCCTCACTAAAGG T3 promoter, forward primer |
| T7 | TAATACGACTCACTATAGGG T7 promoter, forward primer |
| T7 Terminal | GCTAGTTATTGCTCAGCGG T7 terminator, reverse primer |
| Tac promoter | GAGCGGATAACAATTTCACACAGG (Waugh lab) Tac promoter, forward primer |
| tdTomato-Fwd | CTGTTCCTGTACGGCATGG 3' end of tdTomato, forward primer |
| tdTomato-Rev | TCTTTGATGACGGCCATGT 5' end of tdTomato, reverse primer |
| Tet-R | GGCGAGTTTACGGGTTGTTA 5’ end of tetracycline resistance gene, reverse primer |
| TK-pA-R | TTGTCTCCTTCCGTGTTTCA Thymidine kinase polyA, reverse primer |
| Tn7-end | GGGGTGGAAATGGAGTTTTT Bacterial transposon Tn7 |
| TRC-F | CAAGGCTGTTAGAGAGATAATTGGA (Root lab) Human U6 promoter, forward primer |
| Ubx-F | AACTCGTACTTTGAACAGGC Drosophila Ultrabithorax gene, forward primer |
| V5 Reverse | ACCGAGGAGAGGGTTAGGGAT (Invitrogen) V5 epitope, reverse primer |
| WPRE-R | CATAGCGTAAAAGGAGCAACA 5’ end of WPRE, reverse primer |
| XBG-R | GACTCCATTCGGGTGTTC Xenopus beta-globin 3’UTR, reverse primer |
| XEF1a | TTTCGCCCTAACTTCGTGAT Xenopus EF1 alpha enhancer/promoter, forward primer |
| Xpress Forward | TATGGCTAGCATGACTGGT (Invitrogen) Xpress epitope, forward primer |