LOCUS       Exported                6067 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pKR43, complete sequence.
ACCESSION   KF528987
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6067)
  AUTHORS   Rossger K, Charpin-El Hamri G, Fussenegger M.
  TITLE     Reward-based hypertension control by a synthetic brain-dopamine 
            interface
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 110 (45), 18150-18155 (2013)
  PUBMED    24127594
REFERENCE   2  (bases 1 to 6067)
  AUTHORS   Roessger K, Fussenegger M, Charpin-El-Hamri G.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-AUG-2013) BSSE, ETH Zurich, Mattenstrasse 26, Basel, 
            BS 4058, Switzerland
REFERENCE   3  (bases 1 to 6067)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6067
                     /organism="Mammalian expression vector pKR43"
                     /mol_type="other DNA"
                     /db_xref="taxon:1419743"
     regulatory      61..364
                     /regulatory_class="enhancer"
                     /note="CMV enhancer"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      365..568
                     /regulatory_class="promoter"
                     /note="CMV promoter"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             656..2749
                     /codon_start=1
                     /product="dopamine receptor D1/EGFP fusion protein"
                     /label=dopamine receptor D1/EGFP fusion protein
                     /protein_id="AHA35648.1"
                     /translation="MRTLNTSAMDGTGLVVERDFSVRILTACFLSLLILSTLLGNTLVC
                     AAVIRFRHLRSKVTNFFVISLAVSDLLVAVLVMPWKAVAEIAGFWPFGSFCNIWVAFDI
                     MCSTASILNLCVISVDRYWAISSPFRYERKMTPKAAFILISVAWTLSVLISFIPVQLSW
                     HKAKPTSPSDGNATSLAETIDNCDSSLSRTYAISSSVISFYIPVAIMIVTYTRIYRIAQ
                     KQIRRIAALERAAVHAKNCQTTTGNGKPVECSQPESSFKMSFKRETKVLKTLSVIMGVF
                     VCCWLPFFILNCILPFCGSGETQPFCIDSNTFDVFVWFGWANSSLNPIIYAFNADFRKA
                     FSTLLGCYRLCPATNNAIETVSINNNGAAMFSSHHEPRGSISKECNLVYLIPHAVGSSE
                     DLKKEEAAGIARPLEKLSPALSVILDYDTDVSLEKIQPITQNGQHPTGSTGRHHGEQGR
                     GAVHRGGAHPGRAGRRRKRPQVQRVRRGRGRCHLRQADPEVHLHHRQAARALAHPRDHP
                     DLRRAVLQPLPRPHEAARLLQVRHARRLRPGAHHLLQGRRQLQDPRRGEVRGRHPGEPH
                     RAEGHRLQGGRQHPGAQAGVQLQQPQRLYHGRQAEERHQGELQDPPQHRGRQRAARRPL
                     PAEHPHRRRPRAAARQPLPEHPVRPEQRPQREARSHGPAGVRDRRRDHSRHGRAVQVKR
                     PRL"
     regulatory      2007..2016
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2013..2732
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2855..2976
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    2855..2976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2983..3438
     promoter        3465..3569
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3571..3928
                     /regulatory_class="promoter"
     promoter        3571..3928
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3779..3914
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3963..4757
                     /codon_start=1
                     /product="NeoR/KanR"
                     /label=NeoR/KanR
                     /note="resistance to neomycin/kanamycin"
                     /protein_id="AHA35649.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      4989..5036
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    4989..5036
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5365..5953
                     /direction=RIGHT