LOCUS       40924_41505        6145 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSV-Sport-di-FR-HIF, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  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  JOURNAL   Unpublished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 6145)
  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 6145)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6145)
  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            "Unpublished"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..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             83..1732
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    1777..2056
                     /label=5' UTR of mHIF-1alpha (IRES)
                     /note="5' UTR of mHIF-1alpha (IRES)"
     CDS             2057..2989
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(3082..3100)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3189..3228
                     /label=intron SV40 small t-antigen
                     /note="intron SV40 small t-antigen"
     polyA_signal    3487..3621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3858..4436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4610..5467)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5468..5572)
                     /label=AmpR promoter
     promoter        5684..5715
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    5723..5739
                     /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        5774..6103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        6109..6127
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 cgagctctta cgcgtgctag cccgggctcg agatctgcga tctaagtaag cttggcattc
       61 cggtactgtt ggtaaagcca ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc
      121 attctatccg ctggaagatg gaaccgctgg agagcaactg cataaggcta tgaagagata
      181 cgccctggtt cctggaacaa ttgcttttac agatgcacat atcgaggtgg acatcactta
      241 cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa
      301 tacaaatcac agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt
      361 gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga
      421 attgctcaac agtatgggca tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt
      481 gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta ttatcatgga
      541 ttctaaaacg gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc
      601 tcccggtttt aatgaatacg attttgtgcc agagtccttc gatagggaca agacaattgc
      661 actgatcatg aactcctctg gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag
      721 aactgcctgc gtgagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc
      781 ggatactgcg attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact
      841 cggatatttg atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt
      901 tctgaggagc cttcaggatt acaagattca aagtgcgctg ctggtgccaa ccctattctc
      961 cttcttcgcc aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc
     1021 ttctggtggc gctcccctct ctaaggaagt cggggaagcg gttgccaaga ggttccatct
     1081 gccaggtatc aggcaaggat atgggctcac tgagactaca tcagctattc tgattacacc
     1141 cgagggggat gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt
     1201 tgtggatctg gataccggga aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag
     1261 aggtcctatg attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga
     1321 caaggatgga tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt
     1381 catcgttgac cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga
     1441 attggaatcc atcttgctcc aacaccccaa catcttcgac gcaggtgtcg caggtcttcc
     1501 cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat
     1561 gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg
     1621 cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag
     1681 aaaaatcaga gagatcctca taaaggccaa gaagggcgga aagatcgccg tgtaattcta
     1741 gaaagcaggg tggtaacaac gcagagtacg cggggaagcc tcctcggctg aggggacgcg
     1801 aggactgtcc tcgccgccgt cgcgggcagt gtctagccag gccttgacaa gctagccgga
     1861 ggagcgccta ggaacccgag ccggagctca gcgagcgcag cctgcagctc ccgcctcgcc
     1921 gtcccggggg gcgtcccgcc tcccaccccg cctctggact tgtctctttc tccgcgcgcg
     1981 cggacagagc gggcgcttag gcccgagcga gcccgggggc cgccggccgg gaagacaacg
     2041 cgggcaccga ttcgccatgg cttcgaaagt ttatgatcca gaacaaagga aacggatgat
     2101 aactggtccg cagtggtggg ccagatgtaa acaaatgaat gttcttgatt catttattaa
     2161 ttattatgat tcagaaaaac atgcagaaaa tgctgttatt tttttacatg gtaacgcggc
     2221 ctcttcttat ttatggcgac atgttgtgcc acatattgag ccagtagcgc ggtgtattat
     2281 accagacctt attggtatgg gcaaatcagg caaatctggt aatggttctt ataggttact
     2341 tgatcattac aaatatctta ctgcatggtt tgaacttctt aatttaccaa agaagatcat
     2401 ttttgtcggc catgattggg gtgcttgttt ggcatttcat tatagctatg agcatcaaga
     2461 taagatcaaa gcaatagttc acgctgaaag tgtagtagat gtgattgaat catgggatga
     2521 atggcctgat attgaagaag atattgcgtt gatcaaatct gaagaaggag aaaaaatggt
     2581 tttggagaat aacttcttcg tggaaaccat gttgccatca aaaatcatga gaaagttaga
     2641 accagaagaa tttgcagcat atcttgaacc attcaaagag aaaggtgaag ttcgtcgtcc
     2701 aacattatca tggcctcgtg aaatcccgtt agtaaaaggt ggtaaacctg acgttgtaca
     2761 aattgttagg aattataatg cttatctacg tgcaagtgat gatttaccaa aaatgtttat
     2821 tgaatcggac ccaggattct tttccaatgc tattgttgaa ggtgccaaga agtttcctaa
     2881 tactgaattt gtcaaagtaa aaggtcttca tttttcgcaa gaagatgcac ctgatgaaat
     2941 gggaaaatat atcaaatcgt tcgttgagcg agttctcaaa aatgaacaat aagaattccc
     3001 gggtcgacga gctcactagt cggcggccgc tctagaggat ccaagcttac gtacgcgtgc
     3061 atgcgacgtc atagctctct ccctatagtg agtcgtatta taagctagct tgggatcttt
     3121 gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa
     3181 agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactagc tgcatatgct
     3241 tgctgcttga gagttttgct tactgagtat gatttatgaa aatattatac acaggagcta
     3301 gtgattctaa ttgtttgtgt attttagatt cacagtccca aggctcattt caggcccctc
     3361 agtcctcaca gtctgttcat gatcataatc agccatacca catttgtaga ggttttactt
     3421 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3481 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3541 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3601 gtatcttatc atgtctggat cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3661 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3721 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3781 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3841 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3901 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3961 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4021 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     4081 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4201 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4261 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcggc
     4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4561 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4621 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4681 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4741 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4801 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4861 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4921 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4981 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5041 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5101 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5161 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5221 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5281 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5341 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5401 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5461 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5521 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5581 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5641 ataggcgtat acgaggccct ttcactcatt aggcacccca ggctttacac tttatagctt
     5701 ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagcatc
     5761 gatgctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5821 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5881 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5941 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6001 cgtactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6061 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctat ttaggtgaca
     6121 ctatagaagg tacgcctgca ggtac
//
