LOCUS       Exported                8969 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo694, complete sequence.
ACCESSION   MG437033
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8969)
  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 8969)
  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 8969)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8969
                     /organism="Expression vector pLeo694"
                     /mol_type="other DNA"
                     /db_xref="taxon:2172867"
     promoter        3..1184
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     misc_feature    5..1171
                     /label=P-EF1a
                     /note="P-EF1a"
     misc_feature    1200..1204
                     /label=SfiI
                     /note="SfiI"
     regulatory      1237..1246
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1243..1305
                     /label=igk secretion signal
                     /note="igk secretion signal"
     misc_feature    1306
                     /label=scfv
                     /note="scfv"
     misc_feature    1312..1689
                     /label=anti RR120
                     /note="anti RR120"
     misc_feature    1702..2445
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    1975..1977
                     /label=F93A
                     /note="F93A"
     misc_feature    2446..3717
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     regulatory      3809..4036
                     /regulatory_class="terminator"
                     /note="bGH PA"
     polyA_signal    3812..4036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    complement(4085..4104)
                     /label=itr
                     /note="itr"
     promoter        4468..4572
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      4503..4531
                     /regulatory_class="promoter"
                     /note="Amp prom"
     CDS             4573..5433
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    4771..5430
                     /label=AmpR
                     /note="AmpR"
     rep_origin      5528..6210
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      5582..6210
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      5604..6192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    6619..6638
                     /label=itr
                     /note="itr"
     polyA_site      6755..6810
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     CDS             complement(6975..7574)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    complement(6978..7574)
                     /label=PuroR
                     /note="PuroR"
     misc_feature    complement(7575..7634)
                     /label=P2A
                     /note="P2A"
     CDS             complement(7575..7631)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    complement(7641..8339)
                     /label=BFP
                     /note="BFP"
     regulatory      complement(8353..8964)
                     /regulatory_class="promoter"
                     /note="RPBSA"
     regulatory      8353..8964
                     /regulatory_class="promoter"
                     /note="RPBSA"