LOCUS       Exported                6528 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pEF6-FLAG-hPKR-ND, complete sequence.
ACCESSION   LT726883
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6528)
  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  JOURNAL   Unpublished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 6528)
  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 6528)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6528
                     /organism="Mammalian expression vector pEF6-FLAG-hPKR-ND"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5387."
                     /db_xref="taxon:1944998"
     promoter        351..1529
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     regulatory      386..1529
                     /regulatory_class="promoter"
                     /note="hEF1alpha promoter"
     misc_feature    582..1520
                     /label=intron
                     /note="intron"
     intron          582..1520
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1546..1564
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1546..1562
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    1600..1623
                     /label=FLAG epitope
                     /note="FLAG epitope"
     CDS             1600..1623
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1633..2388
                     /codon_start=1
                     /note="unnamed protein product; hPKR-ND"
                     /protein_id="SJL86764.1"
                     /translation="MAGDLSAGFFMEELNTYRQKQGVVLKYQELPNSGPPHDRRFTFQV
                     IIDGREFPEGEGRSKKEAKNAAAKLAVEILNKEKKAVSPLLLTTTNSSEGLSMGNYIGL
                     INRIAQKKRLTVNYEQCASGVHGPEGFHYKCKMGQKEYSIGTGSTKQEAKQLAAKLAYL
                     QILSEETSVKSDYLSSGSFATTCESQSNSLVTSTLASESSSEGDFSADTSEINSNSDSL
                     NSSSLLMNGLRNNQRKAKRSLAPRFDLPD"
     misc_feature    2410..2439
                     /label=Myc epitope
                     /note="Myc epitope"
     CDS             2410..2439
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2455..2472
                     /label=His-tag
                     /note="His-tag"
     CDS             2455..2472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      2501..2725
                     /regulatory_class="terminator"
                     /note="bovine growth hormone polyA"
     polyA_signal    2501..2725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2771..3199
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      3204..3485
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        3213..3542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3393..3528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3442..3517
                     /label=SV40 ori
                     /note="SV40 ori"
     regulatory      3590..3655
                     /regulatory_class="promoter"
                     /note="EM7 promoter"
     promoter        3590..3637
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3656..4054
                     /codon_start=1
                     /note="unnamed protein product; blasticidin"
                     /protein_id="SJL86765.1"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4212..4333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(4212..4286)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      4212..4286
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(4382..4398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4406..4422
                     /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)."
     promoter        complement(4430..4460)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4475..4496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4600..4991)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(4773..5361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5532..6392)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL86766.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6393..6497)
                     /gene="bla"
                     /label=AmpR promoter