LOCUS       Exported                4912 bp ds-DNA    circular SYN 26-7-2013
DEFINITION  Vector with a minimal promoter and NF-kappa-B response element for 
            studying cell signaling using destabilized NanoLuc(R) luciferase.
ACCESSION   JQ513377
VERSION     .
KEYWORDS    pNL3.2.NF-kB-RE[NlucP NF-kB-RE Hygro]
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4912)
  AUTHORS   Promega
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4912
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     protein_bind    33..84
                     /bound_moiety="NF-kappa-B"
                     /note="NF-kappa-B RE"
                     /note="NF-kappa-B response element (5 copies)"
     promoter        116..147
                     /note="minP"
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             180..692
                     /codon_start=1
                     /gene="
                     "
                     /product="NanoLuc(R) luciferase"
                     /note="Nluc"
                     /note="human codon-optimized"
                     /protein_id="
                     "
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             696..815
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /note="hPEST"
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    867..988
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        1182..1539
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      1390..1525
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             1570..2607
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /note="HygR"
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     polyA_signal    2631..2679
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3006..3594)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3794..4654)
                     /codon_start=1
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4759..4807
                     /note="synthetic polyadenylation signal"
     misc_feature    4821..4912
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"