LOCUS       sequence-272192-        3391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5 copies of the NF-kappa-beta DNA binding motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3391)
  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 3391)
  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: TB:5xNF- kappa-beta_RE:MiniP
FEATURES             Location/Qualifiers
     source          1..3391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        267..298
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      368..377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      372..381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(411..430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(412..430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(437..454)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(437..453)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(446..468)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      594..1049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(681..700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     891..912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1127..1231
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1232..1300,1301..2092)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1450..1469)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2263..2851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2752..2771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3005..3022
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3139..3160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3175..3180,3181..3198,3199..3205)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3213..3229
                     /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)."
     primer_bind     3218..3240
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3237..3253
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3237..3253
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3271..3289
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3271..3288
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctcgcatg catgagggga atttccgggg
      181 actttcccgg gaatttgcgg gggactttcc gggaatttcc tccctcgagg atatcaaatc
      241 tggcctcggc ggccaagctt agacactaga gggtatataa tggaagctcg acttccagct
      301 tggcaatccg gtactgttgg taaagccacc aatgcgagac cactcatcgc ccaatcgaat
      361 tcccgcggcc gccatggcgg ccgggagcat gcgacgtcgg gcccaattcg ccctatagtg
      421 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
      481 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      541 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
      601 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      661 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      721 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      781 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccagcgcc
      841 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      901 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
      961 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1021 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     1081 tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1141 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1201 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1261 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1321 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1381 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1441 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1501 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1561 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1621 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1681 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1741 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1801 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1861 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1921 tgataaatct ggagccggtg agcgtgggtc gcgcggtatc attgcagcac tggggccaga
     1981 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2041 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2101 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2161 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2221 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2281 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2341 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2401 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2461 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2521 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2581 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2641 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2701 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2761 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2821 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2881 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2941 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3001 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3061 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3121 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     3181 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     3241 aaacagctat gaccatgatt acgccaagct atttaggtga cactatagaa tactcaagct
     3301 atgcatccaa cgcgttggga gctctcccat atggtcgacc tgcaggcggc cgcgaattca
     3361 ctagtgattg ggcgatgagt ggtctcggga g
//
