LOCUS       Exported                6145 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSV-Sport-di-FR-HIF, complete sequence.
ACCESSION   LT726961
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6145
                     /organism="Mammalian expression vector
                     pSV-Sport-di-FR-HIF"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5202."
                     /db_xref="taxon:1945142"
     regulatory      77..86
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             83..1735
                     /codon_start=1
                     /note="unnamed protein product; LUCm"
                     /protein_id="SJL87064.1"
                     /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..2992
                     /codon_start=1
                     /note="unnamed protein product; rLUC"
                     /protein_id="SJL87065.1"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(3082..3100)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(3084..3100)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    3189..3228
                     /label=intron SV40 small t-antigen
                     /note="intron SV40 small t-antigen"
     regulatory      complement(3465..3561)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3465..3561
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     polyA_signal    3487..3621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3674..4076)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     CDS             complement(4607..5467)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL87066.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5468..5572)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      5663..5720
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     protein_bind    5723..5739
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     regulatory      5765..6046
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        5774..6103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5954..6089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      6003..6078
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        6109..6127
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      6109..6125
                     /regulatory_class="promoter"
                     /note="SP6 promoter"