The pGL3 Luciferase Reporter Vectors provide a basis for the quantitative analysis of factors that potentially regulate mammalian gene expression. These factors may be cis-acting, such as promoters and enhancers, or trans-acting, such as various DNA-binding factors. The backbone of the pGL2 Luciferase Reporter Vectors was redesigned for the pGL3 Vectors for increased expression, and a modified coding region for firefly (Photinus pyralis) luciferase that has been optimized for monitoring transcriptional activity in transfected eukaryotic cells. The assay of this genetic reporter is rapid, sensitive and quantitative. In addition, the Luciferase Reporter Vectors contain numerous features aiding in the structural characterization of the putative regulatory sequences under investigation.

The pGL3-Basic Vector lacks eukaryotic promoter and enhancer sequences, allowing maximum flexibility in cloning putative regulatory sequences. Expression of luciferase activity in cells transfected with this plasmid depends on insertion and proper orientation of a functional promoter upstream from luc+. Potential enhancer elements can also be inserted upstream of the promoter or in the BamH I or Sal I sites downstream of the luc+ gene.

 back Return to this vector's summary.


 
LOCUS       CVU47295     4818 bp    DNA             SYN       26-FEB-1996
DEFINITION  Cloning vector pGL3-Basic firefly luciferase (luc+) gene,
            beta-lactamase (AmpR) gene, complete cds.
ACCESSION   U47295
NID         g1200459
KEYWORDS    .
SOURCE      Cloning vector pGL3-Basic.
  ORGANISM  Cloning vector pGL3-Basic
            artificial sequence; cloning vectors.
REFERENCE   1  (bases 1 to 4818)
  AUTHORS   Groskreutz,D.J. and Schenborn,E.T.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-JAN-1996) D.J. Groskreutz, R&D, Promega Corporation,
            5445 East Cheryl Parkway, Madison, WI 53711, USA
COMMENT
            NCBI gi: 1200459
FEATURES               Location/Qualifiers
     source          1..4818
                     /organism="Cloning vector pGL3-Basic"
                     /note="Luciferase Reporter Vector; plasmid"
     misc_feature    1..58
                     /note="Multiple cloning site"
     CDS             88..1740
                     /gene="luc+"
                     /note="NCBI gi: 1200460"
                     /codon_start=1
                     /product="firefly luciferase"
                     /db_xref="PID:g1200460"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAV
                     APANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQG
                     FQSMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVR
                     FSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRS
                     LQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLP
                     GIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCV
                     RGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVA
                     PAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTT
                     AKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(89..111)
                     /gene="luc+"
                     /note="GLprimer2 sequencing primer binding site"
     polyA_signal    1772..1993
                     /note="SV40 late poly(A) signal"
     primer_bind     complement(2061..2080)
                     /note="RVprimer4 sequencing primer binding site"
     rep_origin      2318
                     /note="ColE1-derived plasmid replication origin"
     CDS             complement(3080..3940)
                     /gene="AmpR"
                     /note="ampicillin resistance;  NCBI gi: 1200461"
                     /codon_start=1
                     /product="beta-lactamase"
                     /db_xref="PID:g1200461"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     rep_origin      4073..4527
                     /note="f1 origin"
     polyA_signal    4658..4811
                     /note="Upstream poly(A) signal"
     primer_bind     4760..4779
                     /note="RVprimer3 sequencing primer binding site"
BASE COUNT     1251 a   1146 c   1159 g   1262 t
ORIGIN      
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctaa gtaagcttgg
       61 cattccggta ctgttggtaa agccaccatg gaagacgcca aaaacataaa gaaaggcccg
      121 gcgccattct atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag
      181 agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc
      241 acttacgctg agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg
      301 ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg
      361 gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa
      421 cgtgaattgc tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag
      481 gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc
      541 atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat
      601 ctacctcccg gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca
      661 attgcactga tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct
      721 catagaactg cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc
      781 attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact
      841 acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag
      901 ctgtttctga ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta
      961 ttctccttct tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa
     1021 attgcttctg gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc
     1081 catctgccag gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt
     1141 acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg
     1201 aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt
     1261 gtgagaggtc ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg
     1321 attgacaagg atggatggct acattctgga gacatagctt actgggacga agacgaacac
     1381 ttcttcatcg ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc
     1441 gctgaattgg aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt
     1501 cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag
     1561 acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag
     1621 ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac
     1681 gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgtaa
     1741 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     1801 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1861 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1921 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     1981 tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc
     2041 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     2101 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     2161 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2221 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2281 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2341 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2401 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2461 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     2521 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2581 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2641 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2701 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2761 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2821 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2881 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2941 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3001 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3061 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3121 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3181 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3241 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3301 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3361 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3421 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3481 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3541 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3601 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3661 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3721 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3781 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3841 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3901 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3961 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4021 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     4081 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4141 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4201 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4261 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4321 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4381 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4441 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4501 ttaacaaaat attaacgttt acaatttccc attcgccatt caggctgcgc aactgttggg
     4561 aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa
     4621 gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt
     4681 acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa
     4741 caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag
     4801 aacatttctc tatcgata
//