LOCUS       sequence-257564-        4541 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 4541)
  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 4541)
  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..4541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    21..69
                     /note="synthetic polyadenylation signal"
     misc_feature    83..174
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     123..142
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        287..318
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     polyA_signal    2003..2227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2351..2939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2431..2450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3054..3156
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3157..3972
                     /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             3157..3972
                     /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(3230..3249)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4086..4514
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4173..4192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4383..4404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacgaattc gtctcaggag aataaaatat ctttattttc attacatctg tgtgttggtt
       61 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
      121 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctcgcatg
      181 catgagggga atttccgggg actttcccgg gaatttgcgg gggactttcc gggaatttcc
      241 tccctcgagg atatcaaatc tggcctcggc ggccaagctt agacactaga gggtatataa
      301 tggaagctcg acttccagct tggcaatccg gtactgttgg taaagccacc aatggagaac
      361 atggagaacg acgagaacat tgtggtggga cccaagccgt tctacccgat cgaggagggt
      421 tccgctggca cccagctgcg caagtacatg gagcgctacg ctaagctggg tgctatcgcc
      481 ttcaccaacg ccgtgaccgg tgtggattac tcctacgccg agtacctgga gaagtcctgc
      541 tgcctcggaa aggccctgca gaactacggt ctggtcgtgg atggccgcat tgccctctgc
      601 tcggagaact gcgaggagtt cttcattccc gtgatcgctg gtctgttcat cggagtcggc
      661 gttgccccaa ccaacgagat ctacaccctg cgcgagctgg tccacagcct gggtatcagc
      721 aagccgacca tcgttttctc gagcaagaag ggcctcgaca aggttatcac cgtgcaaaaa
      781 accgtcacca ccatcaagac catcgttatc ctggattcca aggttgacta ccgcggttac
      841 cagtgcctcg acaccttcat caagcgtaac accccacccg gcttccaagc ctcctccttc
      901 aaaaccgtgg aggttgatcg caaggagcag gtggccctga tcatgaactc ctccggctcc
      961 accggtctgc caaagggcgt gcagctgacc cacgagaaca ccgtcacccg cttctcgcat
     1021 gcccgtgatc ccatctacgg caaccaggtg tccccaggca ccgccgtcct gaccgtcgtg
     1081 cccctccacc acggcttcgg catgttcacc actctcggct acttgatctg cggcttccgt
     1141 gtggtgatgc tgactaagtt cgacgaggag actttcctca agaccctgca agactacaag
     1201 tgcacttacg tgatcctggt gccgaccctg ttcgccatcc tgaacaagag cgagctgttg
     1261 aacaaatacg acctgtcgaa cctggtggag atcgcttccg gcggcgcccc actgtcgaag
     1321 gaagttggcg aggccgttgc tcgccgcttc aacctgcccg gcgtccgtca gggttacggc
     1381 ctgaccgaga ctacctcggc tatcatcatc acccccgagg gcgacgataa gcccggcgct
     1441 tccggcaagg ttgttccact cttcaaagcc aaggtgatcg acctggacac caagaagtcc
     1501 ctgggtccca accgtcgcgg agaagtgtgc gtcaagggcc ccatgctgat gaagggctac
     1561 gtcaacaacc cagaggccac caaggagttg atcgacgagg agggctggct gcacactggt
     1621 gacatcggtt actacgacga agagaagcac ttcttcatcg ttgaccgcct gaagtccttg
     1681 atcaagtaca agggctacca ggttcccccc gccgagctcg agagcgtcct gctgcagcac
     1741 ccctccattt tcgatgccgg agtggccggt gtccccgatc ccgtggctgg cgagctgcca
     1801 ggtgccgtcg tcgtgctgga gtccggcaag aacatgaccg agaaggaagt gatggattac
     1861 gtggcctcgc aagtgtccaa cgccaagcgc ctgcgcggcg gcgtgcgctt cgtggatgag
     1921 gtgcccaagg gcctcaccgg aaagatcgac ggtcgcgcta tccgcgagat cctcaagaag
     1981 cccgtggcca agatgtgagc ttctgtgcct tctagttgcc agccatctgt tgtttgcccc
     2041 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2101 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2161 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2221 tctatggcgc tgtcatgaga cgaattctga cacggttatc cacagaatca ggggataacg
     2281 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2341 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2401 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2461 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2521 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2581 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2641 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2701 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2761 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2821 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2881 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2941 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3001 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg tgatgatcgg
     3061 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     3121 gagttatcga gattttcagg agctaaggaa gctaaaatga gccatattca acgggaaacg
     3181 tcttgctcaa ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     3241 gctcgcgata atgtcgggca atcaggtgcg acaatctacc gattgtatgg gaagcccgat
     3301 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     3361 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     3421 cgtactcctg atgatgcatg gttactcacc actgcgatcc cagggaaaac agcattccag
     3481 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     3541 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcga
     3601 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     3661 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     3721 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3781 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3841 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3901 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     3961 gagtttttct gatttttttt aaggcagtta ttggtgccct taaacgcctg gttgctacgc
     4021 ctgaataagt gataataagc ggatgaatgg cagaaattcg atgataagct gtcaaacatg
     4081 atgttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4141 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4201 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4261 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4321 tgggccagcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4381 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4441 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4501 atttaacgcg aattttaaca aaatattaac gcttacaatt t
//
