LOCUS       Exported                6275 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pYL4, complete sequence.
ACCESSION   MH607407
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  AUTHORS   Liu Y, Charpin-El Hamri G, Ye H, Fussenegger M.
  TITLE     A synthetic free fatty acid-regulated transgene switch in mammalian 
            cells and mice
  JOURNAL   Nucleic Acids Res. (2018) In press
  PUBMED    30219861
REFERENCE   2  (bases 1 to 6275)
  AUTHORS   Liu Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUL-2018) Department of Biosystems Science and 
            Engineering, ETH Zurich, Mattenstrasse 26, basel 4058, Switzerland
REFERENCE   3  (bases 1 to 6275)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6275
                     /organism="Expression vector pYL4"
                     /mol_type="other DNA"
                     /db_xref="taxon:2340780"
     CDS             20..922
                     /codon_start=1
                     /gene="FFAR1"
                     /product="free fatty acid receptor 1"
                     /label=FFAR1
                     /note="G-protein coupled receptor 40"
                     /protein_id="AYA60343.1"
                     /translation="MDLPPQLSFGLYVAAFALGFPLNVLAIRGATAHARLRLTPSLVYA
                     LNLGCSDLLLTVSLPLKAVEALASGAWPLPASLCPVFAVAHFFPLYAGGGFLAALSAGR
                     YLGAAFPLGYQAFRRPCYSWGVCAAIWALVLCHLGLVFGLEAPGGWLDHSNTSLGINTP
                     VNGSPVCLEAWDPASAGPARFSLSLLLFFLPLAITAFCYVGCLRALARSGLTHRRKLRA
                     AWVAGGALLTLLLCVGPYNASNVASFLYPNLGGSWRKLGLITGAWSVVLNPLVTGYLGR
                     GPGLKTVCAARTQGGKSQK"
     gene            20..922
                     /gene="FFAR1"
                     /label=FFAR1
     regulatory      960..1184
                     /regulatory_class="polyA_signal_sequence"
                     /note="BGH polyadenylation sequence"
     polyA_signal    960..1184
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1230..1658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1672..2001
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1852..1987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2068..2862
                     /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"
     polyA_signal    3036..3157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3206..3222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3230..3246
                     /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(3254..3284)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3299..3320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3608..4193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4364..5224)
                     /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"
     promoter        complement(5225..5329)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      5592..6179
                     /regulatory_class="promoter"
                     /note="CMV promoter"
     enhancer        5595..5974
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5975..6178
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        6223..6241
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"