LOCUS       Exported                7755 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo669, complete sequence.
ACCESSION   MG437027
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7755)
  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 7755)
  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 7755)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7755
                     /organism="Expression vector pLeo669"
                     /mol_type="other DNA"
                     /db_xref="taxon:2172861"
     misc_feature    8..216
                     /label=Psv40
                     /note="Psv40"
     promoter        19..215
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      66..201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      241..250
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    247..309
                     /label=igk secretion signal
                     /note="igk secretion signal"
     misc_feature    247..295
                     /label=secrete
                     /note="secrete"
     misc_feature    310..1044
                     /label=scfv
                     /note="scfv"
     misc_feature    1060..1803
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    1333..1335
                     /label=F93A
                     /note="F93A"
     misc_feature    1735..1803
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1804..3075
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    3129..3353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3130..3354
                     /label=BGH PolyA
                     /note="BGH PolyA"
     rep_origin      3399..3827
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3416..3722
                     /label=F1 ori
                     /note="F1 ori"
     promoter        3841..4170
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4021..4156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4237..5031
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4240..5031
                     /label=Kan/neoR
                     /note="Kan/neoR"
     polyA_signal    5205..5326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5375..5391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5399..5415
                     /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(5423..5453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5468..5489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    5718..6388
                     /label=pUC ORI
                     /note="pUC ORI"
     rep_origin      complement(5777..6362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6533..7393)
                     /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    complement(6536..7195)
                     /label=AmpR
                     /note="AmpR"
     promoter        complement(7394..7498)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    complement(7435..7463)
                     /label=Amp promoter
                     /note="Amp promoter"