pSA83 vector (V003523)

LOCUS       40924_38393        5599 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSA83, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5599)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature"; 
            date: "2012"; volume: "487"; issue: "7405"; pages: "123-127"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-FEB-2012) Department of Biosystems Science and Engineering, 
            D-BSSE, ETH Zurich, Mattenstrasse 26, Basel, BS 4058, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             427..1269
                     /codon_start=1
                     /label=d2EYFP
                     /note="EYFP destabilized by residues 422-461 of mouse
                     ornithine decarboxylase, giving an in vivo half-life of ~2 
                     hours"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     polyA_signal    1321..1545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1591..2019
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2033..2362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2429..3220
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3397..3530
                     /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    complement(3591..3607)
                     /label=lac operator
                     /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    complement(3660..3681)
                     /label=CAP binding site
                     /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(4728..5585)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(5586..5599,1..91))
                     /label=AmpR promoter