Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at help@addgene.org. Learn more

reference_icon_book.jpgSequencing 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.

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