LOCUS       Exported                6986 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pGal4-p65S276A, complete sequence.
ACCESSION   LT727142
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6986)
  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  JOURNAL   Unpublished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 6986)
  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 6986)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6986
                     /organism="Mammalian expression vector pGal4-p65S276A"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4647."
                     /db_xref="taxon:1945039"
     misc_feature    1..825
                     /label=SV40 VP1 with internal deletion
                     /note="SV40 VP1 with internal deletion"
     polyA_signal    766..900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(826..900)
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     regulatory      826..900
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     misc_feature    complement(934..1723)
                     /label=3 UTR of hNF-kappaB p65 subunit
                     /note="3 UTR of hNF-kappaB p65 subunit"
     CDS             complement(1724..3379)
                     /codon_start=1
                     /note="unnamed protein product; hNF-kappaB p65 mutant"
                     /protein_id="SJL87583.1"
                     /translation="MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGS
                     IPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAEL
                     CPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQV
                     TVRDPSGRPLRLPPVLPHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKE
                     DIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPADRELS
                     EPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSAS
                     VPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAM
                     VSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNST
                     DPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAP
                     LGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS"
     CDS             complement(1727..2098)
                     /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    complement(2552..2557)
                     /label=S276A
                     /note="S276A"
     misc_feature    complement(3380..3415)
                     /label=linker
                     /note="linker"
     CDS             complement(3416..3853)
                     /codon_start=1
                     /note="unnamed protein product; GALbd"
                     /protein_id="SJL87584.1"
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     regulatory      complement(3997..4178)
                     /regulatory_class="promoter"
                     /note="RSV-LTR"
     promoter        complement(3998..4259)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     regulatory      4575..4856
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        4584..4913
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4764..4899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4813..4888
                     /label=SV40 ori
                     /note="SV40 ori"
     rep_origin      complement(4975..5377)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(5159..5747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5918..6778)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL87585.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6779..6883)
                     /gene="bla"
                     /label=AmpR promoter