LOCUS       Exported                5000 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-FLAG-hUb-K63, complete sequence.
ACCESSION   LT726812
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5000)
  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  JOURNAL   Unpublished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 5000)
  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 5000)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5000
                     /organism="Mammalian expression vector
                     pCAGGS-FLAG-hUb-K63"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5903."
                     /db_xref="taxon:1944841"
     misc_feature    13..36
                     /label=FLAG epitope
                     /note="FLAG epitope"
     CDS             13..36
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             46..276
                     /codon_start=1
                     /note="unnamed protein product; hUBB mutated mature 
                     sequence"
                     /protein_id="SJL86572.1"
                     /translation="MQIFVRTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             46..273
                     /codon_start=1
                     /product="human ubiquitin variant containing Lys-63, with 
                     all other lysines mutated to arginine"
                     /label=ubiquitin-K63
                     /translation="MQIFVRTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     misc_feature    61..63
                     /label=K6R
                     /note="K6R"
     misc_feature    76..78
                     /label=K11R
                     /note="K11R"
     misc_feature    124..126
                     /label=K27R
                     /note="K27R"
     misc_feature    130..132
                     /label=K29R
                     /note="K29R"
     misc_feature    142..144
                     /label=K33R
                     /note="K33R"
     misc_feature    187..189
                     /label=K48R
                     /note="K48R"
     regulatory      286..377
                     /regulatory_class="terminator"
                     /note="3 UTR of rabbit beta-globin, incl. polyA"
     polyA_site      353..408
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    378..734
                     /label=3 flanking region of rabbit beta-globin
                     /note="3 flanking region of rabbit beta-globin"
     primer_bind     complement(769..785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    793..809
                     /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(812..894)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(817..847)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    861..882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        941..1136
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      987..1122
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      1036..1111
                     /label=SV40 ori
                     /note="SV40 ori"
     polyA_signal    1142..1276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(1142..1216)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      1142..1216
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      complement(1331..1732)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(1514..2102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2273..3133)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL86573.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3134..3238)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3273..3652
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      3273..3650
                     /regulatory_class="promoter"
                     /note="hCMV-IE enhancer"
     promoter        3654..3931
                     /label=chicken beta-actin promoter
     regulatory      3656..4997
                     /regulatory_class="promoter"
                     /note="AG promoter"
     intron          3932..4949
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    4025..4949
                     /label=intron AG promoter
                     /note="intron AG promoter"