LOCUS       pSF-CMV-PGK-RLuc        5680 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Dual-promoter mammalian vector for strong constitutive expression of
            a gene together with Renilla luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    pSF-CMV-PGK-RLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  AUTHORS   Oxford Genetics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5680)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     The PGK promoter is typically about 10-fold weaker than the CMV 
            promoter.
FEATURES             Location/Qualifiers
     source          1..5680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..89
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus
                     (Farrell et al., 2002)"
     terminator      complement(94..141)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    complement(158..206)
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     enhancer        287..590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        591..794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    857..942
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     RBS             893..898
                     /label=Shine-Dalgarno ribosome binding site
                     /note="Shine-Dalgarno ribosome binding site"
     misc_signal     903..909
                     /label=Kozak sequence
                     /note="Kozak sequence"
     CDS             907..909
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     misc_feature    961..971
                     /label=stop codons
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     promoter        988..1487
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             1498..2433
                     /label=RLuc
                     /note="Renilla luciferase"
     misc_feature    2444..2454
                     /label=stop codons
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    complement(2486..2607)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      2721..2768
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2798..2934
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2979..3050
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus
                     (Farrell et al., 2002)"
     rep_origin      complement(3276..3864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     RBS             4597..4602
                     /label=Shine-Dalgarno ribosome binding site
                     /note="Shine-Dalgarno ribosome binding site"
     CDS             4610..5401
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      5417..5553
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
       61 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
      121 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
      181 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
      241 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      301 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      361 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      421 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      481 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      541 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
      601 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      661 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      721 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      781 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
      841 ctcgacacac ccgccagcgg ccgctgccaa gcttccgagc tctcgaattc aaaggaggta
      901 cccaccatgg ggtaccgcga tatctacctc gaggtttcta gaagttgtct cctcctgcac
      961 tgactgactg atacaatcga tactaccggg taggggaggc gcttttccca aggcagtctg
     1021 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     1081 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     1141 cgccaccttc tacccctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcat
     1201 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcaaatg gacagcaccg
     1261 ctgagcaatg gaagcgggta ggcccttggg gcagcggcca atagcagctt tgctccttcg
     1321 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
     1381 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attccgcacg cttcaaaagc
     1441 gcacgtctgc cgcgctgttc tcttcttcct catctccggg cctttcggga tcccaacatg
     1501 accagcaaag tgtatgatcc ggaacagcgc aaacgcatga ttaccggccc gcagtggtgg
     1561 gcgcgctgca aacagatgaa cgtgctggat agctttatta actattatga tagcgaaaaa
     1621 catgcggaaa acgcggtgat ttttctgcat ggcaacgcgg cgagcagcta tctgtggcgg
     1681 catgtggtgc cgcatattga accagtggcg cgctgcatta ttccggatct gattggcatg
     1741 ggcaaaagcg gcaaaagcgg caacggcagc tatcgcctgc tggatcatta taaatatctg
     1801 accgcgtggt ttgaactgct gaacctgccg aaaaaaatta tttttgtggg ccatgattgg
     1861 ggcgcgtgcc tggcgtttca ttatagctat gaacatcagg ataaaattaa agcgattgtg
     1921 cacgcggaaa gcgtggtgga tgtgattgaa agctgggatg aatggccgga tattgaagaa
     1981 gatattgcgc tgattaaaag cgaagaaggc gaaaaaatgg tgctggaaaa caactttttt
     2041 gtggaaacca tgctgccgag caaaattatg cgcaaactgg aaccggaaga atttgcggcg
     2101 tatctggaac cgtttaaaga aaaaggcgaa gtgcgccgcc cgaccctgag ctggccgcgc
     2161 gaaattccgc tggtgaaagg cggcaaaccg gatgtggtcc agattgtgcg caactataac
     2221 gcgtatctgc gcgcgagcga tgatctgccg aaaatgttta ttgaaagcga tccgggcttt
     2281 tttagcaacg cgattgtgga aggcgcgaaa aaatttccga acaccgaatt tgtgaaagtg
     2341 aagggcctgc attttagcca ggaagatgcg ccggatgaaa tgggcaaata tattaagagc
     2401 tttgtggaac gcgtgctgaa aaacgaacag ggctaggcta gcttgactga ctgagataca
     2461 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     2521 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2581 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2641 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     2701 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     2761 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     2821 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     2881 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     2941 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     3001 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     3061 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     3121 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     3181 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     3241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4081 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     4141 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     4201 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     4261 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     4321 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat
     4381 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4441 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4501 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4561 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgattgaaca
     4621 agatggattg cacgcaggtt ctccggcggc ttgggtggag aggctattcg gctatgactg
     4681 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4741 tccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4801 agcgcggcta tcgtggctgg cgacgacggg cgttccttgc gcggctgtgc tcgacgttgt
     4861 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4921 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4981 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5041 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5101 gctcgcgcca gccgaactgt tcgccaggct caaggcgtct atgcccgacg gcgaggatct
     5161 cgtcgtgacc cacggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5221 tggattcatc gactgtggcc gtctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5281 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc ttgtgcttta
     5341 cggtatcgcc gcgcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5401 ctgaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     5461 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     5521 tacgtgtcct ccttaccaga aatttatcct taaggtcgtt taaactcgac tctggctcta
     5581 tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc
     5641 atcgaatctc gtcagctatc gtcagcttac ctttttggca
//
