LOCUS       40924_11655        5483 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector for constitutive high-level expression of secreted 
            Gaussia luciferase under control of the CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5483)
  AUTHORS   Thermo Scientific (Pierce)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5483)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        615..633
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             647..1201
                     /codon_start=1
                     /label=hGLuc
                     /note="secreted Gaussia luciferase"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    1227..1338
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1461..1790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        1838..1885
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             1904..2500
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    2633..2766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2810..3667)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3931..4519
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4789..4805
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5261..5277
                     /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    5392..5483
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      601 tggcttatcg aaattaatac gactcactat aggggatcca gccaccatgg gagtcaaagt
      661 tctgtttgcc ctgatctgca tcgctgtggc cgaggccaag cccaccgaga acaacgaaga
      721 cttcaacatc gtggccgtgg ccagcaactt cgcgaccacg gatctcgatg ctgaccgcgg
      781 gaagttgccc ggcaagaagc tgccgctgga ggtgctcaaa gagatggaag ccaatgcccg
      841 gaaagctggc tgcaccaggg gctgtctgat ctgcctgtcc cacatcaagt gcacgcccaa
      901 gatgaagaag ttcatcccag gacgctgcca cacctacgaa ggcgacaaag agtccgcaca
      961 gggcggcata ggcgaggcga tcgtcgacat tcctgagatt cctgggttca aggacttgga
     1021 gcccatggag cagttcatcg cacaggtcga tctgtgtgtg gactgcacaa ctggctgcct
     1081 caaagggctt gccaacgtgc agtgttctga cctgctcaag aagtggctgc cgcaacgctg
     1141 tgcgaccttt gccagcaaga tccagggcca ggtggacaag atcaaggggg ccggtggtga
     1201 ctaagcggcc gcaaaatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1261 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1321 aaatgaggaa attgcatcac aacactcaac cctatctcgg tctattcttt tgatttataa
     1381 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1441 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1501 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1561 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1621 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1681 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1741 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     1801 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     1861 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaccg agtacaagcc
     1921 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     1981 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     2041 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     2101 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     2161 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     2221 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     2281 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     2341 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacat ccgcgccccg
     2401 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     2461 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacacgtgc tacgagattt
     2521 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     2581 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     2641 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2701 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2761 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcattac caatgcttaa
     2821 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2881 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga
     2941 taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa
     3001 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3061 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg
     3121 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3181 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3241 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3301 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3361 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3421 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3481 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3541 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3601 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3661 tactcatatt cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3721 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca
     3781 attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc
     3841 ccttgctcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc
     3901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4081 tctagtgtag ccgtagttag cccaccactt caagaactct gtagcaccgc ctacatacct
     4141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4501 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4561 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4621 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4681 agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac taagcagaag
     4741 gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg
     4801 ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt aatccgcaaa
     4861 taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga aagagcattt
     4921 gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat ttaaccttat
     4981 aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat tgatgaacac
     5041 ctataattaa actattcatc tattatttat gattttttgt atatacaata tttctagttt
     5101 gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca gtttttgata
     5161 tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag atgttatcag
     5221 tatttattat catttagaat aaattttgtg tcgcccttaa ttgtgagcgg ataacaatta
     5281 cgagcttcat gcacagtggc gttgacattg attattgact agcatgttct ttcctgcgtt
     5341 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa taacatacgc
     5401 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     5461 tgcaggtgcc agaacatttc tct
//
