LOCUS       sequence-212838-        5651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter with internal control for NFkB signal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5651)
  AUTHORS   Zhou W, Pal AS, Hsu AY, Gurol T, Zhu X, Wirbisky-Hershberger SE, 
            Freeman JL, Kasinski AL, Deng Q
  TITLE     MicroRNA-223 Suppresses the Canonical NF-kappaB Pathway in Basal 
            Keratinocytes to Dampen Neutrophilic Inflammation.
  JOURNAL   Cell Rep. 2018 Feb 13;22(7):1810-1823. doi: 
            10.1016/j.celrep.2018.01.058.
  PUBMED    29444433
REFERENCE   2  (bases 1 to 5651)
  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"; doi: 
            "10.1016/j.celrep.2018.01"; journalName: "Cell Rep"; date: 
            "2018-02-13- 13"; volume: "22"; issue: "7"; pages: "1810-1823"
COMMENT     Sequence Label: pSI-Check2-hRluc-NFkB-firefly
FEATURES             Location/Qualifiers
     source          1..5651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1688..1736
                     /note="synthetic polyadenylation signal"
     promoter        1846..1877
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1904..1913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1910..3562
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             2869..2877
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3606..3727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3617..3636)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3671..3690
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3860..3964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3965..4033,4034..4825)
                     /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(4183..4202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4996..5584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5485..5504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgagcc cgggaattcg tttaaaccta gagcggccgc
     1681 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgagga
     1741 tctcttatca tgtctggatc caagctaggg gactttccgc ttggggactt tccgctgggg
     1801 actttccgct ggggactttc cgctggggac tttccgcgga gactctagag ggtatataat
     1861 ggaagctcga attccagctt ggcattccgg tactgttggt aaagccacca tggccgatgc
     1921 taagaacatt aagaagggcc ctgctccctt ctaccctctg gaggatggca ccgctggcga
     1981 gcagctgcac aaggccatga agaggtatgc cctggtgcct ggcaccattg ccttcaccga
     2041 tgcccacatt gaggtggaca tcacctatgc cgagtacttc gagatgtctg tgcgcctggc
     2101 cgaggccatg aagaggtacg gcctgaacac caaccaccgc atcgtggtgt gctctgagaa
     2161 ctctctgcag ttcttcatgc cagtgctggg cgccctgttc atcggagtgg ccgtggcccc
     2221 tgctaacgac atttacaacg agcgcgagct gctgaacagc atgggcattt ctcagcctac
     2281 cgtggtgttc gtgtctaaga agggcctgca gaagatcctg aacgtgcaga agaagctgcc
     2341 tatcatccag aagatcatca tcatggactc taagaccgac taccagggct tccagagcat
     2401 gtacacattc gtgacatctc atctgcctcc tggcttcaac gagtacgact tcgtgccaga
     2461 gtctttcgac agggacaaaa ccattgccct gatcatgaac agctctgggt ctaccggcct
     2521 gcctaagggc gtggccctgc ctcatcgcac cgcctgtgtg cgcttctctc acgcccgcga
     2581 ccctattttc ggcaaccaga tcatccccga caccgctatt ctgagcgtgg tgccattcca
     2641 ccacggcttc ggcatgttca ccaccctggg ctacctgatt tgcggctttc gggtggtgct
     2701 gatgtaccgc ttcgaggagg agctgttcct gcgcagcctg caagactaca aaattcagtc
     2761 tgccctgctg gtgccaaccc tgttcagctt cttcgctaag agcaccctga tcgacaagta
     2821 cgacctgtct aacctgcacg agattgcctc tggcggcgcc ccactgtcta aggaggtggg
     2881 cgaagccgtg gccaagcgct ttcatctgcc aggcatccgc cagggctacg gcctgaccga
     2941 gacaaccagc gccattctga ttaccccaga gggcgacgac aagcctggcg ccgtgggcaa
     3001 ggtggtgcca ttcttcgagg ccaaggtggt ggacctggac accggcaaga ccctgggagt
     3061 gaaccagcgc ggcgagctgt gtgtgcgcgg ccctatgatt atgtccggct acgtgaataa
     3121 ccctgaggcc acaaacgccc tgatcgacaa ggacggctgg ctgcactctg gcgacattgc
     3181 ctactgggac gaggacgagc acttcttcat cgtggaccgc ctgaagtctc tgatcaagta
     3241 caagggctac caggtggccc cagccgagct ggagtctatc ctgctgcagc accctaacat
     3301 tttcgacgcc ggagtggccg gcctgcccga cgacgatgcc ggcgagctgc ctgccgccgt
     3361 cgtcgtgctg gaacacggca agaccatgac cgagaaggag atcgtggact atgtggccag
     3421 ccaggtgaca accgccaaga agctgcgcgg cggagtggtg ttcgtggacg aggtgcccaa
     3481 gggcctgacc ggcaagctgg acgcccgcaa gatccgcgag atcctgatca aggctaagaa
     3541 aggcggcaag atcgccgtgt aataattcta gagtcggggc ggccggccgc ttcgagcaga
     3601 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3661 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3721 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3781 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccaggtgg
     3841 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3901 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3961 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4021 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4081 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4141 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4201 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4261 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4321 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4381 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4441 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4501 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4561 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4621 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4681 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4741 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4801 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4861 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4921 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4981 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5041 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5101 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5161 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5221 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5281 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5341 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5401 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5461 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5521 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5581 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5641 catggctcga c
//
