LOCUS       Exported                5599 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSA83, complete sequence.
ACCESSION   JQ624673
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5599)
  AUTHORS   Auslaender S, Auslaender D, Mueller M, Wieland M, Fussenegger M.
  TITLE     Programmable single-cell mammalian biocomputers
  JOURNAL   Nature 487 (7405), 123-127 (2012)
  PUBMED    22722847
REFERENCE   2  (bases 1 to 5599)
  AUTHORS   Auslaender S, Auslaender D, Mueller M, Wieland M, Fussenegger M.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2012) Department of Biosystems Science and 
            Engineering, D-BSSE, ETH Zurich, Mattenstrasse 26, Basel, BS 4058, 
            Switzerland
REFERENCE   3  (bases 1 to 5599)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5599
                     /organism="Mammalian expression vector pSA83"
                     /mol_type="other DNA"
                     /db_xref="taxon:1323710"
     regulatory      complement(28..56)
                     /regulatory_class="promoter"
                     /note="AmpR promoter"
     regulatory      123..315
                     /regulatory_class="promoter"
                     /note="TtgO operator sequence fused to minimal human 
                     cytomegalovirus immediate-early promoter; TtgR1"
     promoter        189..227
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        349..367
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    385..407
                     /label=C/D RNA box
                     /note="C/D RNA box"
     regulatory      421..430
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             427..1272
                     /codon_start=1
                     /gene="d2EYFP"
                     /product="d2EYFP"
                     /label=d2EYFP
                     /protein_id="AGJ84355.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     gene            427..1272
                     /gene="d2EYFP"
                     /label=d2EYFP
     regulatory      1321..1545
                     /regulatory_class="polyA_signal_sequence"
                     /note="BGH polyA"
     polyA_signal    1321..1545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1591..2019
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     regulatory      complement(2024..2367)
                     /regulatory_class="enhancer"
                     /note="simian virus 40 enhancer"
     promoter        2033..2362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2213..2348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2429..3223
                     /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    3397..3518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3567..3583)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3591..3607
                     /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(3615..3645)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3660..3681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3969..4554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4725..5585)
                     /codon_start=1
                     /gene="AmpR"
                     /product="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance protein"
                     /protein_id="AGJ84356.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4725..5585)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(5586..91)
                     /gene="bla"
                     /label=AmpR promoter