LOCUS       40924_41515        5787 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSV-Sport-di-RF, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  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  JOURNAL   Unpublished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 5787)
  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 5787)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5787)
  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            "Unpublished"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..5787
                     /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"
     CDS             1400..3049
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        complement(3105..3123)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3212..3251
                     /label=intron SV40 small t-antigen
                     /note="intron SV40 small t-antigen"
     polyA_signal    3510..3644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3881..4459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4633..5490)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5491..5595)
                     /label=AmpR promoter
     promoter        5707..5738
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    5746..5762
                     /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 cgggtcgaga tctgcgatct aagtaagctt ggcattccgg
     1381 tactgttggt aaagccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt
     1441 ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc
     1501 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc
     1561 tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac
     1621 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg
     1681 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt
     1741 gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca
     1801 aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc
     1861 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc
     1921 cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact
     1981 gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac
     2041 tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga
     2101 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg
     2161 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct
     2221 gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt
     2281 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc
     2341 tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc
     2401 aggtatcagg caaggatatg ggctcactga gactacatca gctattctga ttacacccga
     2461 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt
     2521 ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg
     2581 tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa
     2641 ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat
     2701 cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt
     2761 ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga
     2821 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac
     2881 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg
     2941 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa
     3001 aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagag
     3061 gatccaagct tacgtacgcg tgcatgcgac gtcatagctc tctccctata gtgagtcgta
     3121 ttataagcta gcttgggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
     3181 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
     3241 gtgttaaact agctgcatat gcttgctgct tgagagtttt gcttactgag tatgatttat
     3301 gaaaatatta tacacaggag ctagtgattc taattgtttg tgtattttag attcacagtc
     3361 ccaaggctca tttcaggccc ctcagtcctc acagtctgtt catgatcata atcagccata
     3421 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3481 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3541 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3601 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcctgcat taatgaatcg
     3661 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3721 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3781 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3841 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3901 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3961 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4021 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     4081 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4141 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4201 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4261 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4321 ggacagtatt tggtatctgc ggccagttac cttcggaaaa agagttggta gctcttgatc
     4381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5641 tatcatgaca ttaacctata aaaataggcg tatacgaggc cctttcactc attaggcacc
     5701 ccaggcttta cactttatag cttccggctc gtataatgtg tggaattgtg agcggataac
     5761 aatttcacac aggaaacagc atcgatg
//
