LOCUS       Exported                5790 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCDM8-His-PKR, complete sequence.
ACCESSION   LT727246
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5790)
  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  JOURNAL   Unpublished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 5790)
  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 5790)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5790
                     /organism="Mammalian expression vector pCDM8-His-PKR"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4512."
                     /db_xref="taxon:1944912"
     rep_origin      25..160
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(36..111)
                     /direction=LEFT
                     /label=SV40 ori
                     /note="SV40 ori"
     misc_feature    complement(262..1004)
                     /label=PY
                     /note="PY"
     rep_origin      complement(262..374)
                     /direction=LEFT
                     /label=PY-O
                     /note="PY-O"
     polyA_signal    1003..1137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(1063..1159)
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     regulatory      1063..1159
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     misc_feature    complement(1160..1558)
                     /label=3 UTR of SV40 small t-antigen
                     /note="3 UTR of SV40 small t-antigen"
     CDS             complement(1409..1429)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    complement(1559..1625)
                     /label=intron of SV40 small t-antigen
                     /note="intron of SV40 small t-antigen"
     CDS             complement(1626..1700)
                     /codon_start=1
                     /note="unnamed protein product; part of SV40 small 
                     t-antigen"
                     /protein_id="SJL87866.1"
                     /translation="DLCEGTLLLWCDIIGQTTYRDLKL"
     CDS             complement(1793..3448)
                     /codon_start=1
                     /note="unnamed protein product; hPKR"
                     /protein_id="SJL87867.1"
                     /translation="MAGDLSAGFFMEELNTYRQKQGVVLKYQELPNSGPPHDRRFTFQV
                     IIDGREFPEGEGRSKKEAKNAAAKLAVEILNKEKKAVSPLLLTTTNSSEGLSMGNYIGL
                     INRIAQKKRLTVNYEQCASGVHGPEGFHYKCKMGQKEYSIGTGSTKQEAKQLAAKLAYL
                     QILSEETSVKSDYLSSGSFATTCESQSNSLVTSTLASESSSEGDFSADTSEINSNSDSL
                     NSSSLLMNGLRNNQRKAKRSLAPRFDLPDMKETKYTVDKRFGMDFKEIELIGSGGFGQV
                     FKAKHRIDGKTYVIKRVKYNNEKAEREVKALAKLDHVNIVHYNGCWDGFDYDPETSDDS
                     LESSDYDPENSKNSSRSKTKCLFIQMEFCDKGTLEQWIEKRRGEKLDKVLALELFEQIT
                     KGVDYIHSKKLIHRDLKPSNIFLVDTKQVKIGDFGLVTSLKNDGKRTRSKGTLRYMSPE
                     QISSQDYGKEVDLYALGLILAELLHVCDTAFETSKFFTDLRDGIISDIFDKKEKTLLQK
                     LLSKKPEDRPNTSEILRTLTVWKKSPEKNERHTC"
     misc_feature    complement(3449..3511)
                     /label=H6TH
                     /note="H6TH"
     CDS             complement(3455..3472)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(3482..3499)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3518..3540
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(3603..3621)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(3605..3621)
                     /regulatory_class="promoter"
                     /note="T7"
     regulatory      complement(3666..4249)
                     /regulatory_class="promoter"
                     /note="hCMV"
     promoter        complement(3666..3869)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        3870..4249
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    complement(4404..4606)
                     /label=supF
                     /note="supF"
     tRNA            complement(4408..4587)
                     /product="E. coli amber-suppressor tyrosine tRNA"
                     /label=supF
     rep_origin      4730..5188
                     /label=M13-O
                     /note="M13-O"
     rep_origin      complement(5207..5439)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"