LOCUS       40924_11665        5860 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector for constitutive high-level expression of 
            intracellular green Renilla luciferase under control of the CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5860)
  AUTHORS   Thermo Scientific (Pierce)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5860)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5860
                     /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..1578
                     /codon_start=1
                     /label=Green Renilla Luciferase
                     /note="green-emitting variant of Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGSALAFHYAYEHQDRIKAIVHMESVVDVIESWMGWPDIEEEL
                     ALIKSEEGEKMVLENNFFVETLLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1604..1715
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1838..2167
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2215..2262
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2281..2877
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3010..3143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3187..4044)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4308..4896
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5166..5182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5638..5654
                     /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    5769..5860
                     /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 ccaccatggc cagcaaggtg
      661 tacgaccccg agcagcggaa gcggatgatc accggccctc agtggtgggc tcggtgcaag
      721 cagatgaacg tgctggacag cttcatcaac tactacgaca gcgagaagca cgccgagaac
      781 gccgtgatct tcctgcacgg caacgccacc agcagctacc tgtggcggca cgtggtgccc
      841 cacatcgagc ctgtggccag atgcatcatc cccgacctga tcggcatggg caagagcggc
      901 aagtccggca acggcagcta ccggctgctg gaccactaca agtacctgac cgcttggttt
      961 gagctgctga acctgcccaa gaagatcatc ttcgtcggcc acgactgggg cagcgccctg
     1021 gcctttcact acgcctacga gcaccaggac cggatcaagg ccatcgtgca catggaaagc
     1081 gtggtggacg tgatcgagag ctggatgggc tggcccgaca tcgaggaaga actggccctg
     1141 atcaagagcg aagagggcga gaagatggtg ctggaaaaca acttcttcgt ggaaaccctg
     1201 ctgcccagca agatcatgcg gaagctggaa cccgaagagt tcgccgccta cctggaaccc
     1261 ttcaaagaaa agggcgaagt gcggaggccc accctgagct ggcccagaga gatccccctg
     1321 gtcaagggcg gcaagcccga cgtggtgcag atcgtgcgga actacaacgc ctacctgcgg
     1381 gccagcgacg acctgcctaa gctgttcatc gagagcgacc ccggcttctt cagcaacgcc
     1441 atcgtggaag gcgccaagaa gttccccaac accgagttcg tgaaagtgaa gggcctgcac
     1501 ttcctccagg aagatgcccc cgacgagatg ggcaagtaca tcaagagctt cgtggaacgg
     1561 gtgctgaaga acgagcagtg agcggccgca aaatcagcct cgactgtgcc ttctagttgc
     1621 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1681 actgtccttt cctaataaaa tgaggaaatt gcatcacaac actcaaccct atctcggtct
     1741 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1801 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     1861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1921 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1981 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2041 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2101 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2161 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     2221 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     2281 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     2341 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     2401 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     2461 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     2521 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     2581 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     2641 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     2701 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     2761 gagacatccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     2821 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     2881 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     2941 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3001 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3061 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3121 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3181 tcattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3241 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3301 ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt tatcagcaat
     3361 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3421 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3481 caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3541 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3601 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3661 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3721 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3781 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3841 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3901 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3961 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4021 gacacggaaa tgttgaatac tcatattctt cctttttcaa tattattgaa gcatttatca
     4081 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4141 ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat ttatacctga
     4201 atatggctca taacacccct tgctcatgac caaaatccct taacgtgagt tacgcgcgcg
     4261 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4321 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4381 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4441 ataccaaata ctgttcttct agtgtagccg tagttagccc accacttcaa gaactctgta
     4501 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4561 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4621 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4681 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4741 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4801 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4861 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4921 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4981 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5041 accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga actgccaggc
     5101 atcaaactaa gcagaaggcc cctgacggat ggcctttttg cgtttctaca aactctttct
     5161 gtgttgtaaa acgacggcca gtcttaagct cgggccccct gggcggttct gataacgagt
     5221 aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt
     5281 ttagggaaag agcatttgtc agaatattta agggcgcctg tcactttgct tgatatatga
     5341 gaattattta accttataaa tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt
     5401 tttcgattga tgaacaccta taattaaact attcatctat tatttatgat tttttgtata
     5461 tacaatattt ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa taataaaggg
     5521 aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat ataatacaaa
     5581 ctataagatg ttatcagtat ttattatcat ttagaataaa ttttgtgtcg cccttaattg
     5641 tgagcggata acaattacga gcttcatgca cagtggcgtt gacattgatt attgactagc
     5701 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     5761 ttattaataa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
     5821 gctgtcccca gtgcaagtgc aggtgccaga acatttctct
//
