LOCUS       40924_41510        6196 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSV-Sport-di-RF-UNR, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6196)
  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  JOURNAL   Unpublished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 6196)
  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 6196)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6196)
  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            "Unpublished"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..6196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        345..363
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             398..1330
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    1378..1806
                     /label=5' UTR of UNR (IRES)
                     /note="5' UTR of UNR (IRES)"
     CDS             1809..3458
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        complement(3514..3532)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3621..3660
                     /label=intron SV40 small t-antigen
                     /note="intron SV40 small t-antigen"
     polyA_signal    3919..4053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4290..4868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5042..5899)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5900..6004)
                     /label=AmpR promoter
     promoter        6116..6147
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    6155..6171
                     /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)."
ORIGIN
        1 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
       61 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
      121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
      181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggcgta
      241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctatttag gtgacactat
      361 agaaggtacg cctgcaggta ccggtccgga attcgccatg gcttcgaaag tttatgatcc
      421 agaacaaagg aaacggatga taactggtcc gcagtggtgg gccagatgta aacaaatgaa
      481 tgttcttgat tcatttatta attattatga ttcagaaaaa catgcagaaa atgctgttat
      541 ttttttacat ggtaacgcgg cctcttctta tttatggcga catgttgtgc cacatattga
      601 gccagtagcg cggtgtatta taccagacct tattggtatg ggcaaatcag gcaaatctgg
      661 taatggttct tataggttac ttgatcatta caaatatctt actgcatggt ttgaacttct
      721 taatttacca aagaagatca tttttgtcgg ccatgattgg ggtgcttgtt tggcatttca
      781 ttatagctat gagcatcaag ataagatcaa agcaatagtt cacgctgaaa gtgtagtaga
      841 tgtgattgaa tcatgggatg aatggcctga tattgaagaa gatattgcgt tgatcaaatc
      901 tgaagaagga gaaaaaatgg ttttggagaa taacttcttc gtggaaacca tgttgccatc
      961 aaaaatcatg agaaagttag aaccagaaga atttgcagca tatcttgaac cattcaaaga
     1021 gaaaggtgaa gttcgtcgtc caacattatc atggcctcgt gaaatcccgt tagtaaaagg
     1081 tggtaaacct gacgttgtac aaattgttag gaattataat gcttatctac gtgcaagtga
     1141 tgatttacca aaaatgttta ttgaatcgga cccaggattc ttttccaatg ctattgttga
     1201 aggtgccaag aagtttccta atactgaatt tgtcaaagta aaaggtcttc atttttcgca
     1261 agaagatgca cctgatgaaa tgggaaaata tatcaaatcg ttcgttgagc gagttctcaa
     1321 aaatgaacaa taagaattcc cgggtcgacg agctcactag tcggcggccg ctctagatgc
     1381 tgcttatggc ggcgctggag agggggcgct gagctgttgg gtatgaagtg taacagaaca
     1441 gactttacca cctgaaactg ctgcttcaag ttcagatcag gcaaggaaca aacctcgtaa
     1501 caactaacaa gaccaaagaa gagtacactt aagttgaaga cacaacactt gatctgaaac
     1561 aagaagtttg tgcctactca acagctttga aagagcactt cccaacgctg ctagtagtct
     1621 ttgttttctt cagtgctgta ctgtgagatt gcccggtaca gcagcagttg tattctttat
     1681 tagcttggta gatcattttc tctcgctctt ttttttaata ctagcaactt tcatcctttg
     1741 aaacgtgtgc tgaaaaagaa gaatcagcaa atactactga aagtgcaata tttgagtatc
     1801 actgcgccat ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg
     1861 aagatggaac cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg
     1921 gaacaattgc ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg
     1981 aaatgtccgt tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa
     2041 tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta
     2101 tcggagttgc agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta
     2161 tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga
     2221 acgtgcaaaa aaagctccca atcatccaaa aaattattat catggattct aaaacggatt
     2281 accagggatt tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg
     2341 aatacgattt tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact
     2401 cctctggatc tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga
     2461 gattctcgca tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt
     2521 taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat
     2581 gtggatttcg agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc
     2641 aggattacaa gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa
     2701 gcactctgat tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc
     2761 ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc
     2821 aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag ggggatgata
     2881 aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata
     2941 ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta
     3001 tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc
     3061 tacattctgg agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc
     3121 tgaagtctct gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct
     3181 tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg
     3241 gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga
     3301 tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt
     3361 ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga
     3421 tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta attctagagg atccaagctt
     3481 acgtacgcgt gcatgcgacg tcatagctct ctccctatag tgagtcgtat tataagctag
     3541 cttgggatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
     3601 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
     3661 gctgcatatg cttgctgctt gagagttttg cttactgagt atgatttatg aaaatattat
     3721 acacaggagc tagtgattct aattgtttgt gtattttaga ttcacagtcc caaggctcat
     3781 ttcaggcccc tcagtcctca cagtctgttc atgatcataa tcagccatac cacatttgta
     3841 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3901 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3961 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4021 aaactcatca atgtatctta tcatgtctgg atcctgcatt aatgaatcgg ccaacgcgcg
     4081 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     4501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4741 ggtatctgcg gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4801 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4861 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4921 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4981 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5041 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5101 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5161 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5221 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5281 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5341 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5401 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5461 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5521 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5581 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5641 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5701 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5761 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5821 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5881 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5941 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6001 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6061 taacctataa aaataggcgt atacgaggcc ctttcactca ttaggcaccc caggctttac
     6121 actttatagc ttccggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
     6181 ggaaacagca tcgatg
//
