LOCUS       sequence-272280-        4780 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 4780)
  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 4780)
  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..4780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      297..752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(384..403)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     594..615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    778..826
                     /note="synthetic polyadenylation signal"
     misc_feature    840..931
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     880..899
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1092..1123
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             1157..2809
                     /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(1223..1241)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2817..3024
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3153..3741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3233..3252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4147..4166)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(4158..4780,1..193)
                     /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             join(4158..4780,1..193)
                     /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(4231..4250)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
       61 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
      121 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
      181 tgagtttttc taatcactag accaatgtta cacatatata ctttagattg atttaaaact
      241 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgtgttacgc
      301 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      361 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      421 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      481 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccagc
      541 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      601 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      661 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      721 gaattttaac aaaatattaa cgcttacaat tttaacagat ctcgtctcag tcaggagaat
      781 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta
      841 acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc
      901 agtgcaagtg caggtgccag aacatttctc tcgctgtcag gaggtctaga ctgccgtcta
      961 gacttaacag tacgtctaga ctcggccgtc tagacttagt acgtctagac tcggtatgtc
     1021 tagactgacc cgtctagacg gatcctccct cgaggatatc aaatctggcc tcggcggcca
     1081 agcttagaca ctagagggta tataatggaa gctcgacttc cagcttggca atccggtact
     1141 gttggtaaag ccaccaatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta
     1201 tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct
     1261 ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga
     1321 gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa
     1381 tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc
     1441 gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct
     1501 caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa
     1561 aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa
     1621 aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg
     1681 ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat
     1741 catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc
     1801 ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac
     1861 tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta cactcggata
     1921 tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag
     1981 gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt
     2041 cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg
     2101 tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg
     2161 tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg
     2221 ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga
     2281 tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc
     2341 tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga
     2401 tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt
     2461 tgaccgcctg aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga
     2521 atccatcttg ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga
     2581 tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga
     2641 aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg
     2701 agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat
     2761 cagagagatc ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat gagcttctgt
     2821 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2881 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2941 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3001 agagaatagc aggcatgctg gggagcggcg ctaatgtgag acgagatctt gacacggtta
     3061 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3121 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3181 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3241 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3301 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3361 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3421 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3481 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3541 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3601 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3661 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3721 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3781 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3841 tagatccttt tgtgagctgt tggatgggga tcgcctgaat cgccccatca tccagccaga
     3901 aagtgaggga gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga
     3961 acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca
     4021 actcagcaaa agttcgattt attcaacaaa gccacgttgt gtctcaaaat ctctggtgtt
     4081 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     4141 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctca aggccgcgat
     4201 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     4261 aatcaggtgc gacaatctac cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     4321 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     4381 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     4441 ggttactcac cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg
     4501 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     4561 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg actcgctcag gcgcaatcac
     4621 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     4681 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     4741 ctcatggtga tttctcactt gataacctta tttttgacga
//
