LOCUS       Exported                3400 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo665, complete sequence.
ACCESSION   MG437024
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3400)
  AUTHORS   Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
  TITLE     Generalized extracellular molecule sensor platform for programming 
            cellular behavior
  JOURNAL   Nat. Chem. Biol. 14 (7), 723-729 (2018)
  PUBMED    29686358
REFERENCE   2  (bases 1 to 3400)
  AUTHORS   Scheller L, Strittmatter T.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 3400)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3400
                     /organism="Expression vector pLeo665"
                     /mol_type="other DNA"
                     /db_xref="taxon:2172858"
     protein_bind    15..285
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     misc_feature    15..33
                     /label=tetO
                     /note="tetO"
     misc_feature    57..75
                     /label=tetO
                     /note="tetO"
     misc_feature    141..159
                     /label=tetO
                     /note="tetO"
     misc_feature    183..201
                     /label=tetO
                     /note="tetO"
     misc_feature    225..243
                     /label=tetO
                     /note="tetO"
     misc_feature    267..285
                     /label=tetO
                     /note="tetO"
     misc_feature    318..442
                     /label=CMVmin
                     /note="CMVmin"
     promoter        318..355
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    464..526
                     /label=IgK-Secretion-Signal
                     /note="IgK-Secretion-Signal"
     CDS             467..526
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="ETDTLLLWVLLLWVPGSTGD"
     misc_feature    545..1057
                     /label=nluc
                     /note="nluc"
     CDS             545..1057
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    1087..1221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1091..1282
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyA"
     rep_origin      complement(1565..2247)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      complement(1583..2171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2565..3200)
                     /label=AmpR
                     /note="AmpR"
     CDS             complement(2638..3016)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5-AACGTT-3 (Sato et al., 1996)"
     promoter        complement(3201..3305)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3242..3270)
                     /regulatory_class="promoter"
                     /note="Amp prom"