LOCUS       Exported                6306 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pFLAG-p65-S536A, complete sequence.
ACCESSION   LT726820
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  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  JOURNAL   Unpublished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 6306)
  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 6306)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6306
                     /organism="Mammalian expression vector pFLAG-p65-S536A"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 6091."
                     /db_xref="taxon:1945030"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      318..901
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    935..958
                     /label=FLAG epitope
                     /note="FLAG epitope"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             962..2614
                     /codon_start=1
                     /note="unnamed protein product; hNF-kappaB p65 mutant"
                     /protein_id="SJL86589.1"
                     /translation="DELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSI
                     PGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELC
                     PDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVT
                     VRDPSGRPLRLPPVLPHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKED
                     IEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSE
                     PMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASV
                     PKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMV
                     SALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTD
                     PAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPL
                     GAPGLPNGLLSGDEDFSAIADMDFSALLSQISS"
     CDS             2240..2611
                     /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
                     EDFSAIADMDFSALLSQISS"
     misc_feature    2564..2566
                     /label=S536A
                     /note="S536A"
     regulatory      2644..3269
                     /regulatory_class="terminator"
                     /note="GH1 polyA"
     polyA_signal    2647..3269
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     regulatory      3289..3570
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        3298..3627
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3478..3613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3527..3602
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        complement(3666..3684)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(3668..3684)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     primer_bind     complement(3698..3714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3722..3738
                     /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(3741..3824)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(3746..3776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3791..3812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3916..4318)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(4100..4688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4859..5719)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL86590.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5720..5824)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5851..6306
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"