LOCUS       40924_11650        5482 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector for constitutive high-level expression of secreted 
            Gaussia-Dura luciferase under control of the CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5482)
  AUTHORS   Thermo Scientific (Pierce)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5482)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5482
                     /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             646..1200
                     /codon_start=1
                     /label=Gaussia-Dura Luc
                     /note="enhanced variant of secreted Gaussia luciferase"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    1226..1337
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1460..1789
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        1837..1884
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             1903..2499
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    2632..2765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2809..3666)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3930..4518
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4788..4804
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5260..5276
                     /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    5391..5482
                     /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 aggggatccg ccaccatggg agtcaaagtt
      661 ctgtttgccc tgatctgcat cgctgtggcc gaggccaagc ccaccgagaa caacgaagac
      721 ttcaacatcg tggccgtggc cagcaacttc gcgaccacgg atctcgatgc tgaccgcggg
      781 aagttgcccg gcaagaagct gccgctggag gtgctcaaag agttggaagc caatgcccgg
      841 aaagctggct gcaccagggg ctgtctgatc tgcctgtccc acatcaagtg cacgcccaag
      901 atgaagaagt tcatcccagg acgctgccac acctacgaag gcgacaaaga gtccgcacag
      961 ggcggcatag gcgaggcgat cgtcgacatt cctgagattc ctgggttcaa ggacttggag
     1021 cccttggagc agttcatcgc acaggtcgat ctgtgtgtgg actgcacaac tggctgcctc
     1081 aaagggcttg ccaacgtgca gtgttctgac ctgctcaaga agtggctgcc gcaacgctgt
     1141 gcgacctttg ccagcaagat ccagggccag gtggacaaga tcaagggggc cggtggtgac
     1201 taagcggccg caaaatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1261 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1321 aatgaggaaa ttgcatcaca acactcaacc ctatctcggt ctattctttt gatttataag
     1381 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1441 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     1501 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     1561 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     1621 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     1681 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     1741 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     1801 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     1861 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatgaccga gtacaagccc
     1921 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     1981 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     2041 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     2101 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     2161 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     2221 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     2281 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     2341 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacatc cgcgccccgc
     2401 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     2461 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacacgtgct acgagatttc
     2521 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     2581 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     2641 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2701 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     2761 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcattacc aatgcttaat
     2821 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2881 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagcg ctgcgatgat
     2941 accgcgagaa ccacgctcac cggctccgga tttatcagca ataaaccagc cagccggaag
     3001 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3061 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccatcgc
     3121 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3181 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3241 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3301 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3361 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3421 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3481 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3541 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3601 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3661 actcatattc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3721 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa
     3781 ttaaccaatt ctgaacatta tcgcgagccc atttatacct gaatatggct cataacaccc
     3841 cttgctcatg accaaaatcc cttaacgtga gttacgcgcg cgtcgttcca ctgagcgtca
     3901 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3961 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4021 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4081 ctagtgtagc cgtagttagc ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4141 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4201 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4261 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4321 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4381 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4441 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4501 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4561 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4621 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4681 gtgagcgagg aagcggaagg cgagagtagg gaactgccag gcatcaaact aagcagaagg
     4741 cccctgacgg atggcctttt tgcgtttcta caaactcttt ctgtgttgta aaacgacggc
     4801 cagtcttaag ctcgggcccc ctgggcggtt ctgataacga gtaatcgtta atccgcaaat
     4861 aacgtaaaaa cccgcttcgg cgggtttttt tatgggggga gtttagggaa agagcatttg
     4921 tcagaatatt taagggcgcc tgtcactttg cttgatatat gagaattatt taaccttata
     4981 aatgagaaaa aagcaacgca ctttaaataa gatacgttgc tttttcgatt gatgaacacc
     5041 tataattaaa ctattcatct attatttatg attttttgta tatacaatat ttctagtttg
     5101 ttaaagagaa ttaagaaaat aaatctcgaa aataataaag ggaaaatcag tttttgatat
     5161 caaaattata catgtcaacg ataatacaaa atataataca aactataaga tgttatcagt
     5221 atttattatc atttagaata aattttgtgt cgcccttaat tgtgagcgga taacaattac
     5281 gagcttcatg cacagtggcg ttgacattga ttattgacta gcatgttctt tcctgcgtta
     5341 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat aacatacgct
     5401 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     5461 gcaggtgcca gaacatttct ct
//
