LOCUS       40924_9936        8253 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pcDNA3-di-RF-ODC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8253)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8253)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 8253)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8253)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             960..1892
                     /codon_start=1
                     /label=Rluc
                     /note="luciferase from the anthozoan coelenterate Renilla 
                     reniformis (sea pansy)"
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNDE"
     misc_feature    1904..2058
                     /label=5' UTR of ODC (IRES)
                     /note="5' UTR of ODC (IRES)"
     CDS             2061..3710
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     3715..3731
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3805..3823)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3849..4073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4119..4547
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4561..4891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4958..5749
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5926..6059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6096..6112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6120..6136)
                     /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)."
     promoter        complement(6144..6174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6189..6210)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6498..7086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7260..8117)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8118..8222)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcttac gcgtgctagc ccgggctcga gatctgcgat ctaagtaagc ttagccacca
      961 tgacttcgaa agtttatgat ccagaacaaa ggaaacggat gataactggt ccgcagtggt
     1021 gggccagatg taaacaaatg aatgttcttg attcatttat taattattat gattcagaaa
     1081 aacatgcaga aaatgctgtt atttttttac atggtaacgc ggcctcttct tatttatggc
     1141 gacatgttgt gccacatatt gagccagtag cgcggtgtat tataccagac cttattggta
     1201 tgggcaaatc aggcaaatct ggtaatggtt cttataggtt acttgatcat tacaaatatc
     1261 ttactgcatg gtttgaactt cttaatttac caaagaagat catttttgtc ggccatgatt
     1321 ggggtgcttg tttggcattt cattatagct atgagcatca agataagatc aaagcaatag
     1381 ttcacgctga aagtgtagta gatgtgattg aatcatggga tgaatggcct gatattgaag
     1441 aagatattgc gttgatcaaa tctgaagaag gagaaaaaat ggttttggag aataacttct
     1501 tcgtggaaac catgttgcca tcaaaaatca tgagaaagtt agaaccagaa gaatttgcag
     1561 catatcttga accattcaaa gagaaaggtg aagttcgtcg tccaacatta tcatggcctc
     1621 gtgaaatccc gttagtaaaa ggtggtaaac ctgacgttgt acaaattgtt aggaattata
     1681 atgcttatct acgtgcaagt gatgatttac caaaaatgtt tattgaatcg gacccaggat
     1741 tcttttccaa tgctattgtt gaaggtgcca agaagtttcc taatactgaa tttgtcaaag
     1801 taaaaggtct tcatttttcg caagaagatg cacctgatga aatgggaaaa tatatcaaat
     1861 cgttcgttga gcgagttctc aaaaatgatg aataatatct agagctggcc tgcggcgcct
     1921 gggcgctctg agattgtcac tgctgttcca agggcacacg cagagggatt tggaattcct
     1981 ggagagttgc ctttgtgaga agctggaaat atttctttca attccatctc ttagttttcc
     2041 ataggaacat caagaaatcc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat
     2101 tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg
     2161 ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg
     2221 ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata
     2281 caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg
     2341 gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat
     2401 tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc
     2461 aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt
     2521 ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc
     2581 ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac
     2641 tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa
     2701 ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg
     2761 atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg
     2821 gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc
     2881 tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct
     2941 tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt
     3001 ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc
     3061 caggtatcag gcaaggatat gggctcactg agactacatc agctattctg attacacccg
     3121 agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg
     3181 tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag
     3241 gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca
     3301 aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca
     3361 tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat
     3421 tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg
     3481 acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga
     3541 cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg
     3601 gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa
     3661 aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg taattctaga
     3721 actagtggat cccccgggct gcaggaattc tgcagatatc catcacactg gcggccgctc
     3781 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     3841 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3901 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3961 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4021 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4081 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4141 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4201 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4261 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4321 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4381 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4441 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4501 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4561 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag
     4621 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4681 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4741 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4801 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4861 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4921 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4981 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5041 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5101 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5161 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5221 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5281 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5341 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5401 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5461 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5521 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5581 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5641 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5701 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5761 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5821 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5881 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5941 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6001 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6061 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6121 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6181 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6241 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6301 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6361 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6421 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6481 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6541 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6601 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6661 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     6721 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6781 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6841 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6901 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6961 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7021 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7081 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7141 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7201 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7261 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7321 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7381 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7441 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7501 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7561 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7621 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7681 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7741 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7801 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7861 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7921 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7981 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8041 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8101 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8161 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8221 cgcacatttc cccgaaaagt gccacctgac gtc
//
