LOCUS       Exported                4744 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pEGFP-NLS, complete sequence.
ACCESSION   LT726832
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4744)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4744)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 4744)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4744
                     /organism="Mammalian expression vector pEGFP-NLS"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 6459."
                     /db_xref="taxon:1945022"
     regulatory      1..568
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             648..670
                     /codon_start=1
                     /note="unnamed protein product; SV40 large T NLS"
                     /protein_id="SJL86627.1"
                     /translation="PKKKRKV"
     CDS             648..668
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      684..693
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             690..1409
                     /codon_start=1
                     /note="unnamed protein product; EGFP"
                     /protein_id="SJL86628.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      complement(1510..1606)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      1510..1606
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     polyA_signal    1532..1653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1660..2115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      2116..2211
                     /regulatory_class="promoter"
                     /note="ampicillin promoter"
     promoter        2142..2246
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2212..2246
                     /label=5 UTR of amp, incl. RBS
                     /note="5 UTR of amp, incl. RBS"
     promoter        2248..2605
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      2267..2548
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     rep_origin      2456..2591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2505..2580
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             2640..3434
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL86629.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      3625..3694
                     /regulatory_class="terminator"
                     /note="HSV-TK polyA"
     polyA_signal    3666..3713
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4042..4630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4412..4738
                     /label=pMB1 ori
                     /note="pMB1 ori"