LOCUS       Exported                6049 bp ds-DNA    circular SYN 27-7-2013
DEFINITION  Vector with a minimal promoter and an NF-kappa-B response element 
            for studying cell signaling using destabilized luciferase.
ACCESSION   EU581860
VERSION     .
KEYWORDS    pGL4.32[luc2P NF-kB-RE Hygro]
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6049)
  AUTHORS   Promega
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6049
                     /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..1829
                     /codon_start=1
                     /product="firefly luciferase"
                     /note="luciferase"
                     /note="synthetic luc2 version of the luciferase gene"
                     /protein_id="
                     "
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             1833..1952
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /note="hPEST"
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2004..2125
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        2319..2676
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2527..2662
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2707..3744
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /note="HygR"
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     polyA_signal    3768..3816
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4143..4731)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4931..5791)
                     /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    5896..5944
                     /note="synthetic polyadenylation signal"
     misc_feature    5958..6049
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"