LOCUS       40924_11621        6586 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector for constitutive high-level expression of secreted 
            Cypridina luciferase under control of the CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6586)
  AUTHORS   Thermo Scientific (Pierce)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6586)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6586
                     /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..2304
                     /codon_start=1
                     /label=CLuc
                     /note="secreted Cypridina luciferase"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     polyA_signal    2330..2441
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2564..2893
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2941..2988
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3007..3603
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3736..3869
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3913..4770)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5034..5622
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5892..5908
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6364..6380
                     /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    6495..6586
                     /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 ccaccatgaa gaccctgatc
      661 ctggccgtgg ccctggtgta ctgcgccacc gtgcactgcc aggactgccc ctacgagccc
      721 gaccccccca ataccgtgcc taccagctgc gaggccaaag agggcgaatg catcgacagc
      781 agctgcggca cctgtacccg ggacatcctg agcgacggcc tgtgcgagaa caagcccggc
      841 aagacctgct gccggatgtg ccagtacgtg atcgagtgcc gggtggaagc cgccggatgg
      901 ttccggacct tctacggcaa gcggttccag tttcaggaac ccggcaccta cgtcctgggc
      961 cagggcacaa agggcggcga ctggaaggtg tccatcaccc tggaaaacct ggacggcacc
     1021 aagggcgccg tgctgaccaa gaccagactg gaagtggccg gcgacatcat cgatatcgcc
     1081 caggccaccg agaaccccat caccgtgaac ggcggagccg accccatcat tgccaacccc
     1141 tacaccatcg gcgaagtgac aatcgctgtg gtggaaatgc ccggcttcaa tatcaccgtc
     1201 atcgagttct tcaagctgat cgtgatcgac atcctgggcg gcagaagcgt gcggatcgcc
     1261 cccgataccg ccaacaaggg catgatcagc ggcctgtgtg gcgacctgaa gatgatggaa
     1321 gataccgact tcaccagcga ccccgagcag ctggccatcc agcccaagat caaccaggaa
     1381 tttgacggct gccccctgta cggcaacccc gacgacgtgg cctactgcaa gggcctgctg
     1441 gaaccctaca aggacagctg cagaaacccc atcaacttct actactacac catcagctgc
     1501 gccttcgccc ggtgcatggg cggagatgag cgcgctagcc acgtgctgct ggactacaga
     1561 gagacatgcg ccgctcccga gacacggggc acatgtgtgc tgagcggcca caccttctac
     1621 gacaccttcg acaaggccag ataccagttc cagggcccct gcaaagaaat cctgatggcc
     1681 gccgactgct tctggaacac ctgggacgtg aaggtgtccc accggaacgt ggacagctat
     1741 accgaggtgg aaaaagtgcg gatcagaaag cagagcaccg tggtcgagct gattgtggac
     1801 ggcaagcaga tcctcgtggg cggcgaggcc gtgtccgtgc cctacagcag ccagaacacc
     1861 agcatctact ggcaggacgg cgacatcctg accaccgcca tcctgcctga ggccctggtg
     1921 gtcaagttca acttcaagca gctgctggtg gtgcacatcc gggacccctt cgacggcaag
     1981 acatgcggca tctgcggcaa ctacaaccag gacttcagcg acgacagctt cgacgccgag
     2041 ggcgcctgcg acctgacccc taatcctccc ggctgcaccg aggaacagaa gcccgaggcc
     2101 gagcggctgt gcaatagcct gttcgccggc cagagcgacc tggaccagaa atgcaacgtg
     2161 tgccacaagc ccgaccgggt ggaacggtgt atgtacgagt actgcctgcg gggccagcag
     2221 ggcttctgcg atcacgcctg ggagttcaag aaagagtgct acatcaagca cggcgacacc
     2281 ctggaagtgc ccgacgagtg caagtgagcg gccgcaaaat cagcctcgac tgtgccttct
     2341 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2401 actcccactg tcctttccta ataaaatgag gaaattgcat cacaacactc aaccctatct
     2461 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2521 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     2581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2641 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2701 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2761 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2821 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2881 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     2941 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     3001 taaaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     3061 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     3121 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     3181 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     3241 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     3301 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     3361 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     3421 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     3481 ttcctggaga catccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     3541 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     3601 gcctgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3661 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3721 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3781 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3841 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     3901 tcatggtcat taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4021 atctggcccc agcgctgcga tgataccgcg agaaccacgc tcaccggctc cggatttatc
     4081 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4141 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4201 tttgcgcaac gttgttgcca tcgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4321 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4441 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4501 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4561 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4621 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4741 aagggcgaca cggaaatgtt gaatactcat attcttcctt tttcaatatt attgaagcat
     4801 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4861 aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga gcccatttat
     4921 acctgaatat ggctcataac accccttgct catgaccaaa atcccttaac gtgagttacg
     4981 cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5041 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5101 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5161 gcgcagatac caaatactgt tcttctagtg tagccgtagt tagcccacca cttcaagaac
     5221 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5281 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5341 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5401 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5461 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5521 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5581 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5641 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5701 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5761 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaggcgagag tagggaactg
     5821 ccaggcatca aactaagcag aaggcccctg acggatggcc tttttgcgtt tctacaaact
     5881 ctttctgtgt tgtaaaacga cggccagtct taagctcggg ccccctgggc ggttctgata
     5941 acgagtaatc gttaatccgc aaataacgta aaaacccgct tcggcgggtt tttttatggg
     6001 gggagtttag ggaaagagca tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat
     6061 atatgagaat tatttaacct tataaatgag aaaaaagcaa cgcactttaa ataagatacg
     6121 ttgctttttc gattgatgaa cacctataat taaactattc atctattatt tatgattttt
     6181 tgtatataca atatttctag tttgttaaag agaattaaga aaataaatct cgaaaataat
     6241 aaagggaaaa tcagtttttg atatcaaaat tatacatgtc aacgataata caaaatataa
     6301 tacaaactat aagatgttat cagtatttat tatcatttag aataaatttt gtgtcgccct
     6361 taattgtgag cggataacaa ttacgagctt catgcacagt ggcgttgaca ttgattattg
     6421 actagcatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6481 cattagttat taataacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     6541 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctct
//
