LOCUS       Exported                6575 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMSK1-E-D565A, complete sequence.
ACCESSION   LT727062
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6575)
  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  JOURNAL   Unpublished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 6575)
  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 6575)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6575
                     /organism="Shuttle vector pMSK1-E-D565A"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4982."
                     /db_xref="taxon:1945226"
     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"
     rep_origin      complement(262..374)
                     /direction=LEFT
                     /label=polyoma ori
                     /note="polyoma ori"
     polyA_signal    1006..1140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(1066..1162)
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     regulatory      1066..1162
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     misc_feature    complement(1163..1561)
                     /label=3 UTR of SV40 small t-antigen
                     /note="3 UTR of SV40 small t-antigen"
     CDS             complement(1412..1432)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    complement(1562..1628)
                     /label=intron of SV40 small t-antigen
                     /note="intron of SV40 small t-antigen"
     intron          1562..1627
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(1629..1703)
                     /codon_start=1
                     /note="unnamed protein product; part of SV40 small 
                     t-antigen"
                     /protein_id="SJL87321.1"
                     /translation="DLCEGTLLLWCDIIGQTTYRDLKL"
     promoter        1711..1729
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      1711..1727
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     misc_feature    complement(1871..1909)
                     /label=E-tag
                     /note="E-tag"
     CDS             complement(1919..4324)
                     /codon_start=1
                     /note="unnamed protein product; hMSK1 mutant"
                     /protein_id="SJL87322.1"
                     /translation="MEEEGGSSGGAAGTSADGGDGGEQLLTVKHELRTANLTGHAEKVG
                     IENFELLKVLGTGAYGKVFLVRKISGHDTGKLYAMKVLKKATIVQKAKTTEHTRTERQV
                     LEHIRQSPFLVTLHYAFQTETKLHLILDYINGGELFTHLSQRERFTEHEVQIYVGEIVL
                     ALEHLHKLGIIYRDIKLENILLDSNGHVVLTDFGLSKEFVADETERAYSFCGTIEYMAP
                     DIVRGGDSGHDKAVDWWSLGVLMYELLTGASPFTVDGEKNSQAEISRRILKSEPPYPQE
                     MSALAKDLIQRLLMKDPKKRLGCGPRDADEIKEHLFFQKINWDDLAAKKVPAPFKPVIR
                     DELDVSNFAEEFTEMDPTYSPAALPQSSEKLFQGYSFVAPSILFKRNAAVIDPLQFHMG
                     VERPGVTNVARSAMMKDSPFYQHYDLDLKDKPLGEGSFSICRKCVHKKSNQAFAVKIIS
                     KRMEANTQKEITALKLCEGHPNIVKLHEVFHDQLHTFLVMELLNGGELFERIKKKKHFS
                     ETEASYIMRKLVSAVSHMHDVGVVHRDLKPENLLFTDENDNLEIKIIAFGFARLKPPDN
                     QPLKTPCFTLHYAAPELLNQNGYDESCDLWSLGVILYTMLSGQVPFQSHDRSLTCTSAV
                     EIMKKIKKGDFSFEGEAWKNVSQEAKDLIQGLLTVDPNKRLKMSGLRYNEWLQDGSQLS
                     SNPLMTPDILGSSGAAVHTCVKATFHAFNKYKREGFCLQNVDKAPLAKRRKMKKTSTST
                     ETRSSSSESSHSSSSHSHGKTTPTKTLQPSNPADSNNPETLFQFSDSVA"
     misc_feature    complement(2630..2632)
                     /label=D565A
                     /note="D565A"
     promoter        complement(4388..4406)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(4390..4406)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     regulatory      complement(4451..5034)
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     promoter        complement(4451..4654)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        4655..5034
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    complement(5189..5391)
                     /label=supF
                     /note="supF"
     tRNA            complement(5193..5372)
                     /product="E. coli amber-suppressor tyrosine tRNA"
                     /label=supF
     rep_origin      5515..5973
                     /label=M13 ori
                     /note="M13 ori"
     rep_origin      complement(5992..6224)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"