LOCUS       Exported                6358 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pEYFP-p65, complete sequence.
ACCESSION   LT726794
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6358)
  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  JOURNAL   Unpublished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 6358)
  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 6358)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6358
                     /organism="Mammalian expression vector pEYFP-p65"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 8224."
                     /db_xref="taxon:1945025"
     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"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1329
                     /codon_start=1
                     /note="unnamed protein product; EYFP"
                     /protein_id="SJL86501.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1363..3015
                     /codon_start=1
                     /note="unnamed protein product; hNF-kappaB p65"
                     /protein_id="SJL86502.1"
                     /translation="MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGS
                     IPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAEL
                     CPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQV
                     TVRDPSGRPLRLPPVLPHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKE
                     DIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELS
                     EPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSAS
                     VPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAM
                     VSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNST
                     DPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAP
                     LGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS"
     CDS             2644..3015
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (OShea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     regulatory      complement(3124..3220)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3124..3220
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     polyA_signal    3146..3267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3274..3729)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      3730..3825
                     /regulatory_class="promoter"
                     /note="ampicillin resistance promoter"
     promoter        3756..3860
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3826..3860
                     /label=5 UTR of amp, incl. RBS
                     /note="5 UTR of amp, incl. RBS"
     promoter        3862..4219
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      3881..4162
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     rep_origin      4070..4205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4119..4194
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             4254..5048
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL86503.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      5239..5308
                     /regulatory_class="terminator"
                     /note="HSV-TK polyA"
     polyA_signal    5280..5327
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5656..6244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6026..6352
                     /label=pMB1 ori
                     /note="pMB1 ori"