LOCUS       sequence-257565-        4793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SMAD2 FLuc luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4793)
  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 4793)
  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: 7xSMAD_RE::FLuc
FEATURES             Location/Qualifiers
     source          1..4793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(296..315)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             307..1122
                     /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             307..1122
                     /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(380..399)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1226..1681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1313..1332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1523..1544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1707..1755
                     /note="synthetic polyadenylation signal"
     misc_feature    1769..1860
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1809..1828
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2021..2052
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             2086..3738
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2152..2170)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3746..3970
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4095..4683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4175..4194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgagctgtt ggatggggat cgcctgaatc gccccatcat ccagccagaa agtgagggag
       61 ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt
      121 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
      181 gttcgattta ttcaacaaag ccacgttgtg tctcaaaatc tctggtgtta cattgcacaa
      241 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
      301 ggtgttatga gccatattca acgggaaacg tcttgctcaa ggccgcgatt aaattccaac
      361 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
      421 acaatctacc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
      481 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
      541 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
      601 actgcgatcc cagggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
      661 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
      721 tgtcctttta acagcgatcg cgtatttcga ctcgctcagg cgcaatcacg aatgaataac
      781 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
      841 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
      901 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
      961 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     1021 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     1081 aataaattgc agtttcattt gatgctcgat gagtttttct aatcactaga ccaatgttac
     1141 acatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1201 atcctttttg ataatctcat gtgttacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1261 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1321 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1381 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1441 tagggtgatg gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg
     1501 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1561 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1621 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1681 ttaacagatc tcgtctcagt caggagaata aaatatcttt attttcatta catctgtgtg
     1741 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
     1801 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
     1861 cgctgtcagg aggtctagac tgccgtctag acttaacagt acgtctagac tcggccgtct
     1921 agacttagta cgtctagact cggtatgtct agactgaccc gtctagacgg atcctccctc
     1981 gaggatatca aatctggcct cggcggccaa gcttagacac tagagggtat ataatggaag
     2041 ctcgacttcc agcttggcaa tccggtactg ttggtaaagc caccaatgga agacgccaaa
     2101 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
     2161 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
     2221 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
     2281 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
     2341 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     2401 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
     2461 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
     2521 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     2581 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
     2641 ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct
     2701 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
     2761 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     2821 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     2881 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     2941 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     3001 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     3061 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     3121 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     3181 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     3241 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     3301 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     3361 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     3421 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     3481 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     3541 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     3601 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     3661 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     3721 ggaaagatcg ccgtgtaatg agcttctgtg ccttctagtt gccagccatc tgttgtttgc
     3781 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3841 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3901 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3961 ggctctatgg cgctaatgtg agacgagatc ttgacacggt tatccacaga atcaggggat
     4021 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4081 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4141 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4201 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4261 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4321 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4381 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4441 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4501 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4561 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4621 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4681 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4741 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct ttt
//
