LOCUS       Exported                7087 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pRcRSVp65S276C, complete sequence.
ACCESSION   LT727083
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7087)
  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  JOURNAL   Unpublished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 7087)
  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 7087)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7087
                     /organism="Mammalian expression vector pRcRSVp65S276C"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5086."
                     /db_xref="taxon:1945057"
     promoter        340..602
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     regulatory      421..603
                     /regulatory_class="promoter"
                     /note="RSV-LTR"
     CDS             700..2355
                     /codon_start=1
                     /note="unnamed protein product; mutated hNF-kappaB p65 
                     subunit"
                     /protein_id="SJL87382.1"
                     /translation="MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGS
                     IPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAEL
                     CPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQV
                     TVRDPSGRPLRLPPVLPHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKE
                     DIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPCDRELS
                     EPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSAS
                     VPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAM
                     VSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNST
                     DPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAP
                     LGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS"
     misc_feature    1519..1527
                     /label=S276C
                     /note="S276C"
     CDS             1981..2352
                     /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"
     misc_feature    2356..2541
                     /label=3 UTR of hNF-kappaB p65 subunit
                     /note="3 UTR of hNF-kappaB p65 subunit"
     regulatory      2588..2812
                     /regulatory_class="terminator"
                     /note="BGH polyA"
     polyA_signal    2588..2812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2858..3371
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2858..3313
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      3390..3672
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        3399..3729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3580..3715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3629..3704
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             3796..4590
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL87383.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4764..4898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(4764..4838)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      4764..4838
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(4930..4946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4954..4970
                     /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)."
     regulatory      complement(4973..5056)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(4978..5008)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5023..5044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5148..5550)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(5332..5920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6091..6951)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87384.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6952..7056)
                     /gene="bla"
                     /label=AmpR promoter