LOCUS       40924_11740        6575 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector for constitutive high-level expression of 
            intracellular red firefly luciferase under control of the CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6575)
  AUTHORS   Thermo Scientific (Pierce)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6575)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..367
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        368..571
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        616..634
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             647..2290
                     /codon_start=1
                     /label=Red Firefly Luciferase
                     /note="intracellular red firefly luciferase"
                     /translation="MENMENDENIVVGPKPFYPIEEGSAGTQLRKYMERYAKLGAIAFT
                     NAVTGVDYSYAEYLEKSCCLGKALQNYGLVVDGRIALCSENCEEFFIPVIAGLFIGVGV
                     APTNEIYTLRELVHSLGISKPTIVFSSKKGLDKVITVQKTVTTIKTIVILDSKVDYRGY
                     QCLDTFIKRNTPPGFQASSFKTVEVDRKEQVALIMNSSGSTGLPKGVQLTHENTVTRFS
                     HARDPIYGNQVSPGTAVLTVVPFHHGFGMFTTLGYLICGFRVVMLTKFDEETFLKTLQD
                     YKCTYVILVPTLFAILNKSELLNKYDLSNLVEIASGGAPLSKEVGEAVARRFNLPGVRQ
                     GYGLTETTSAIIITPEGDDKPGASGKVVPLFKAKVIDLDTKKSLGPNRRGEVCVKGPML
                     MKGYVNNPEATKELIDEEGWLHTGDIGYYDEEKHFFIVDRLKSLIKYKGYQVPPAELES
                     VLLQHPSIFDAGVAGVPDPVAGELPGAVVVLESGKNMTEKEVMDYVASQVSNAKRLRGG
                     VRFVDEVPKGLTGKIDGRAIREILKKPVAKM"
     polyA_signal    2319..2430
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2553..2882
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2930..2977
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2996..3592
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3725..3858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3902..4759)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5023..5611
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5881..5897
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6353..6369
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    6484..6575
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 aactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
       61 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      121 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      181 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      241 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      301 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      361 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      421 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      481 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      541 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      601 ctggcttatc gaaattaata cgactcacta taggggatcc gccaccatgg aaaatatgga
      661 aaacgacgag aacatcgtgg tgggccccaa gcccttctac cccatcgagg aaggcagcgc
      721 cggcacccag ctgcggaagt acatggaaag atacgccaag ctgggcgcca ttgccttcac
      781 caacgccgtg accggcgtgg actacagcta cgccgagtac ctggaaaaga gctgctgcct
      841 gggcaaggct ctgcagaact acggcctggt ggtggacggc cggatcgccc tgtgcagcga
      901 gaactgcgag gaattcttca tccccgtgat cgccggcctg ttcatcggcg tgggcgtggc
      961 tcccaccaac gagatctaca ccctgcggga gctggtgcac agcctgggca tcagcaagcc
     1021 caccatcgtg ttcagcagca agaagggcct ggacaaagtc atcaccgtgc agaaaaccgt
     1081 gaccaccatc aagaccatcg tgatcctgga cagcaaggtg gactaccggg gctaccagtg
     1141 cctggacacc ttcatcaagc ggaacacccc ccctggcttc caggccagca gcttcaagac
     1201 cgtggaggtg gaccggaaag aacaggtggc cctgatcatg aacagcagcg gcagcaccgg
     1261 cctgcccaag ggcgtgcagc tgacccacga gaacaccgtg acccggttca gccacgccag
     1321 ggaccccatc tacggcaacc aggtgtcccc cggcaccgcc gtgctgaccg tggtgccctt
     1381 ccaccacggc ttcggcatgt tcaccaccct gggctacctg atctgcggct tccgggtggt
     1441 gatgctgacc aagttcgacg aggaaacctt cctgaaaacc ctgcaggact acaagtgcac
     1501 ctacgtgatt ctggtgccca ccctgttcgc catcctgaac aagagcgagc tgctgaacaa
     1561 gtacgacctg agcaacctgg tggagatcgc cagcggcgga gcccccctga gcaaagaagt
     1621 gggagaggcc gtcgccaggc ggttcaatct gcccggcgtg cggcagggct acggcctgac
     1681 cgagacaacc agcgccatca tcatcacccc cgagggcgac gacaagcctg gagccagcgg
     1741 caaggtggtg cccctgttca aggccaaagt gatcgacctg gacaccaaga agagcctggg
     1801 ccccaacaga cggggcgaag tgtgcgtgaa gggccccatg ctgatgaagg gctacgtgaa
     1861 caaccccgag gccaccaaag agctgatcga cgaagagggc tggctgcaca ccggcgacat
     1921 cggctactac gacgaagaga agcacttctt catcgtggac cggctgaaga gcctgatcaa
     1981 gtacaagggc tatcaggtgc cccctgccga gctggaaagc gtcctgctgc agcaccccag
     2041 catcttcgac gccggcgtgg ccggggtgcc agatcctgtg gccggcgagc tgcctggcgc
     2101 cgtggtggtg ctggaatccg gcaagaacat gaccgagaaa gaagtgatgg actacgtcgc
     2161 cagccaggtg tccaacgcca agcggctgag aggcggcgtg agattcgtgg acgaagtgcc
     2221 aaagggcctg accggcaaga tcgacggcag ggccatccgg gagatcctga agaaacccgt
     2281 ggccaagatg tgatgagcgg ccgcaaaatc agcctcgact gtgccttcta gttgccagcc
     2341 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2401 cctttcctaa taaaatgagg aaattgcatc acaacactca accctatctc ggtctattct
     2461 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2521 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2581 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2641 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2701 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2761 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2821 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2881 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     2941 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatgac
     3001 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3061 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3121 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3181 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3241 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3301 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3361 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3421 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3481 atccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3541 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacacgt
     3601 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3661 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3721 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3781 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3841 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcatt
     3901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4021 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
     4081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4201 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4741 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
     4861 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     4921 gctcataaca ccccttgctc atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt
     4981 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5041 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5101 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5161 aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact ctgtagcacc
     5221 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5281 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5341 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5401 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5461 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5521 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5581 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5641 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5701 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5761 gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc caggcatcaa
     5821 actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc tttctgtgtt
     5881 gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa cgagtaatcg
     5941 ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg ggagtttagg
     6001 gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata tatgagaatt
     6061 atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt tgctttttcg
     6121 attgatgaac acctataatt aaactattca tctattattt atgatttttt gtatatacaa
     6181 tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata aagggaaaat
     6241 cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat acaaactata
     6301 agatgttatc agtatttatt atcatttaga ataaattttg tgtcgccctt aattgtgagc
     6361 ggataacaat tacgagcttc atgcacagtg gcgttgacat tgattattga ctagcatgtt
     6421 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     6481 aataacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     6541 ccccagtgca agtgcaggtg ccagaacatt tctct
//
