LOCUS       sequence-272264-        4538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NFKB RedFirefly luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4538)
  AUTHORS   Sarrion-Perdigones A, Gonzalez Y, Venken KJT
  TITLE     Rapid and Efficient Synthetic Assembly of Multiplex Luciferase 
            Reporter Plasmids for the Simultaneous Monitoring of Up to Six 
            Cellular Signaling Pathways.
  JOURNAL   Curr Protoc Mol Biol. 2020 Jun;131(1):e121. doi: 10.1002/cpmb.121.
  PUBMED    32539183
REFERENCE   2  (bases 1 to 4538)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr Protoc
            Mol Biol."; date: "2020-06"; volume: "131(1):e121. doi"; pages: " 
            10.1002/cpmb.121"
COMMENT     Sequence Label: 5xNF-kb_RE::RedF
FEATURES             Location/Qualifiers
     source          1..4538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1317..1541
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1665..2250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1745..1764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2365..2467
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2468..3283
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2468..3283
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2541..2560)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3397..3852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3484..3503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3694..3715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3873..3921
                     /note="synthetic polyadenylation signal"
     misc_feature    3935..4026
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3975..3994
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4139..4170
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
ORIGIN
        1 cctgcgcgag ctggtccaca gcctgggtat cagcaagccg accatcgttt tctcgagcaa
       61 gaagggcctc gacaaggtta tcaccgtgca aaaaaccgtc accaccatca agaccatcgt
      121 tatcctggat tccaaggttg actaccgcgg ttaccagtgc ctcgacacct tcatcaagcg
      181 taacacccca cccggcttcc aagcctcctc cttcaaaacc gtggaggttg atcgcaagga
      241 gcaggtggcc ctgatcatga actcctccgg ctccaccggt ctgccaaagg gcgtgcagct
      301 gacccacgag aacaccgtca cccgcttctc gcatgcccgt gatcccatct acggcaacca
      361 ggtgtcccca ggcaccgccg tcctgaccgt cgtgcccctc caccacggct tcggcatgtt
      421 caccactctc ggctacttga tctgcggctt ccgtgtggtg atgctgacta agttcgacga
      481 ggagactttc ctcaagaccc tgcaagacta caagtgcact tacgtgatcc tggtgccgac
      541 cctgttcgcc atcctgaaca agagcgagct gttgaacaaa tacgacctgt cgaacctggt
      601 ggagatcgct tccggcggcg ccccactgtc gaaggaagtt ggcgaggccg ttgctcgccg
      661 cttcaacctg cccggcgtcc gtcagggtta cggcctgacc gagactacct cggctatcat
      721 catcaccccc gagggcgacg ataagcccgg cgcttccggc aaggttgttc cactcttcaa
      781 agccaaggtg atcgacctgg acaccaagaa gtccctgggt cccaaccgtc gcggagaagt
      841 gtgcgtcaag ggccccatgc tgatgaaggg ctacgtcaac aacccagagg ccaccaagga
      901 gttgatcgac gaggagggct ggctgcacac tggtgacatc ggttactacg acgaagagaa
      961 gcacttcttc atcgttgacc gcctgaagtc cttgatcaag tacaagggct accaggttcc
     1021 ccccgccgag ctcgagagcg tcctgctgca gcacccctcc attttcgatg ccggagtggc
     1081 cggtgtcccc gatcccgtgg ctggcgagct gccaggtgcc gtcgtcgtgc tggagtccgg
     1141 caagaacatg accgagaagg aagtgatgga ttacgtggcc tcgcaagtgt ccaacgccaa
     1201 gcgcctgcgc ggcggcgtgc gcttcgtgga tgaggtgccc aagggcctca ccggaaagat
     1261 cgacggtcgc gctatccgcg agatcctcaa gaagcccgtg gccaagatgt gagcttctgt
     1321 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1381 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1441 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1501 agacaatagc aggcatgctg gggatgcggt gggctctatg gcgctgtcat gagacgaatt
     1561 ctgacacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2101 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2161 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2221 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2281 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2341 atcttcacct agatcctttt gtgatgatcg gcacgtaaga ggttccaact ttcaccataa
     2401 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     2461 agctaaaatg agccatattc aacgggaaac gtcttgctca aggccgcgat taaattccaa
     2521 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2581 gacaatctac cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2641 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2701 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2761 cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2821 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2881 ttgtcctttt aacagcgatc gcgtatttcg actcgctcag gcgcaatcac gaatgaataa
     2941 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3001 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3061 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3121 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3181 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3241 gaataaattg cagtttcatt tgatgctcga tgagtttttc tgattttttt taaggcagtt
     3301 attggtgccc ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg
     3361 gcagaaattc gatgataagc tgtcaaacat gatgttacgc gccctgtagc ggcgcattaa
     3421 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3481 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3541 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3601 aaaaacttga ttagggtgat ggttcacgta gtgggccagc gccctgatag acggtttttc
     3661 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3721 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3781 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3841 cgcttacaat tttaacgaat tcgtctcagg agaataaaat atctttattt tcattacatc
     3901 tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa
     3961 cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat
     4021 ttctctcgca tgcatgaggg gaatttccgg ggactttccc gggaatttgc gggggacttt
     4081 ccgggaattt cctccctcga ggatatcaaa tctggcctcg gcggccaagc ttagacacta
     4141 gagggtatat aatggaagct cgacttccag cttggcaatc cggtactgtt ggtaaagcca
     4201 ccaatggaga acatggagaa cgacgagaac attgtggtgg gacccaagcc gttctacccg
     4261 atcgaggagg gttccgctgg cacccagctg cgcaagtaca tggagcgcta cgctaagctg
     4321 ggtgctatcg ccttcaccaa cgccgtgacc ggtgtggatt actcctacgc cgagtacctg
     4381 gagaagtcct gctgcctcgg aaaggccctg cagaactacg gtctggtcgt ggatggccgc
     4441 attgccctct gctcggagaa ctgcgaggag ttcttcattc ccgtgatcgc tggtctgttc
     4501 atcggagtcg gcgttgcccc aaccaacgag atctacac
//
