LOCUS       pSF-CMV-PGK-FLuc        6394 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Dual-promoter mammalian vector for strong constitutive expression of
            a gene together with firefly luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    pSF-CMV-PGK-FLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6394)
  AUTHORS   Oxford Genetics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6394)
  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..6394
                     /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=Shinbe-Dalgarno ribosome binding site
                     /note="Shinbe-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..3147
                     /label=luciferase
                     /note="firefly luciferase"
     misc_feature    3158..3168
                     /label=stop codons
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    complement(3200..3321)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      3435..3482
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3512..3648
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3693..3764
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus
                     (Farrell et al., 2002)"
     rep_origin      complement(3990..4578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     RBS             5311..5316
                     /label=Shine-Dalgarno ribosome binding site
                     /note="Shine-Dalgarno ribosome binding site"
     CDS             5324..6115
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      6131..6267
                     /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 gaagatgcca aaaacattaa gaaaggccca gcgccattct acccactcga agacgggacc
     1561 gctggcgagc agctgcataa agccatgaag cgctacgccc tggtgcccgg caccatcgcc
     1621 tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga gatgagcgtt
     1681 cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat cgtggtgtgt
     1741 agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat cggtgtggct
     1801 gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat gggcatcagc
     1861 cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa cgtgcaaaag
     1921 aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta ccagggcttc
     1981 caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga gtacgacttc
     2041 gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag tagtggcagt
     2101 accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg attcagtcat
     2161 gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct gagcgtggtg
     2221 ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg cggctttcgg
     2281 gtcgtgctca tgtaccgctt cgaggaagag ctattcttgc gcagcttgca agactataag
     2341 attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag cactctcatc
     2401 gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc gctcagcaag
     2461 gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca gggctacggc
     2521 ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa gcctggcgca
     2581 gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac cggtaagaca
     2641 ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat gagcggctac
     2701 gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct gcatagcggc
     2761 gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct gaagagcctg
     2821 atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct gctgcaacac
     2881 cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg cgagctgccc
     2941 gccgcagtcg tcgtgctgga acacggtaaa accatgaccg agaaggagat cgtggactat
     3001 gtggccagcc aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt cgtggacgag
     3061 gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat tctcattaag
     3121 gccaagaagg gcggcaagat cgccgtctag gctagcttga ctgactgaga tacagcgtac
     3181 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3241 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3301 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3361 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     3421 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3481 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     3541 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     3601 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     3661 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     3721 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     3781 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     3841 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     3901 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     3961 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4021 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4081 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4141 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4201 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4261 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4321 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4381 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4441 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4501 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4561 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4621 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4681 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4741 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4801 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     4861 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     4921 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     4981 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     5041 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc
     5101 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     5161 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     5221 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     5281 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgattg aacaagatgg
     5341 attgcacgca ggttctccgg cggcttgggt ggagaggcta ttcggctatg actgggcaca
     5401 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgtccggt
     5461 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     5521 gctatcgtgg ctggcgacga cgggcgttcc ttgcgcggct gtgctcgacg ttgtcactga
     5581 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5641 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5701 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5761 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5821 gccagccgaa ctgttcgcca ggctcaaggc gtctatgccc gacggcgagg atctcgtcgt
     5881 gacccacggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5941 catcgactgt ggccgtctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6001 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttccttgtgc tttacggtat
     6061 cgccgcgccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgacc
     6121 gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     6181 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     6241 tcctccttac cagaaattta tccttaaggt cgtttaaact cgactctggc tctatcgaat
     6301 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     6361 tctcgtcagc tatcgtcagc ttaccttttt ggca
//
