LOCUS       sequence-359618-        3878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SMAD GreenRenilla luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3878)
  AUTHORS   Sarrion-Perdigones A, Gonzalez Y, Venken KJT
  TITLE     Synthetic Assembly DNA Cloning of Multiplex Hextuple Luciferase 
            Reporter Plasmids.
  JOURNAL   Methods Mol Biol. 2022;2524:409-432. doi: 
            10.1007/978-1-0716-2453-1_32.
  PUBMED    35821490
REFERENCE   2  (bases 1 to 3878)
  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: "Methods Mol
            Biol."; date: "2022"; volume: "2524:409-432. doi"; pages: " 
            10.1007/978-1-0716-2453-1_32"
COMMENT     Sequence Label: 7xSMAD_RE::GrRenilla
FEATURES             Location/Qualifiers
     source          1..3878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    25..73
                     /note="synthetic polyadenylation signal"
     misc_feature    87..178
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     127..146
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        339..370
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     polyA_signal    1344..1568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1688..2276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1768..1787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2391..2493
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2494..3309
                     /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             2494..3309
                     /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(2567..2586)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3423..3851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3510..3529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3720..3741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacgatatc gtctcatggc ggagaataaa atatctttat tttcattaca tctgtgtgtt
       61 ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa
      121 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctcg
      181 ctgtcaggag gtctagactg ccgtctagac ttaacagtac gtctagactc ggccgtctag
      241 acttagtacg tctagactcg gtatgtctag actgacccgt ctagacggat cctccctcga
      301 ggatatcaaa tctggcctcg gcggccaagc ttagacacta gagggtatat aatggaagct
      361 cgacttccag cttggcaatc cggtactgtt ggtaaagcca ccaatggctt ccaaagtcta
      421 cgacccagaa caaaggaaga ggatgattac tggaccgcag tggtgggcaa ggtgcaagca
      481 gatgaatgta ctcgatagtt ttatcaatta ttacgactcc gaaaaacacg cggaaaatgc
      541 tgtcatattc ctgcacggga acgccactag cagctacttg tggcggcacg tagtccccca
      601 catagagcct gttgcccgat gtattattcc cgatctcatt ggaatgggca aatcaggcaa
      661 atctgggaac ggttcttacc gcctgctcga tcactataaa tatttgaccg cttggtttga
      721 gctgcttaat ctccctaaaa aaataatatt cgttggacac gactggggct cagctctggc
      781 gttccactac gcatacgagc atcaagaccg aataaaagca atagtacata tggagtcagt
      841 tgttgacgtc atagagtcat ggatgggttg gccagatatt gaagaggaat tggccctcat
      901 caagagtgag gagggggaaa aaatggtcct ggaaaataat tttttcgtgg aaaccttgct
      961 cccgagtaaa ataatgcgaa aacttgaacc cgaagagttt gctgcttatt tggagccttt
     1021 caaggaaaag ggggaagtaa gacgaccaac cctttcttgg ccgagggaga tacctcttgt
     1081 taagggtgga aagcccgacg tagtccagat agtccgaaat tacaacgcgt acctgagagc
     1141 tagtgatgac ctccctaagt tgtttatcga gtcagaccct gggttcttca gcaacgcaat
     1201 tgtagaagga gcgaaaaaat ttccaaatac agaatttgtg aaggttaagg ggctccactt
     1261 tttgcaagag gacgcccccg atgagatggg aaaatacatt aaatctttcg tagagcgcgt
     1321 ccttaagaat gaacagtgag cttctgtgcc ttctagttgc cagccatctg ttgtttgccc
     1381 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1441 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1501 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     1561 ctctatggcg cttgagacga tatctgacac ggttatccac agaatcaggg gataacgcag
     1621 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1681 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1741 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1801 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1861 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1921 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1981 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2041 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2101 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2161 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2221 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2281 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2341 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgtga tgatcggcac
     2401 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
     2461 ttatcgagat tttcaggagc taaggaagct aaaatgagcc atattcaacg ggaaacgtct
     2521 tgctcaaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2581 cgcgataatg tcgggcaatc aggtgcgaca atctaccgat tgtatgggaa gcccgatgcg
     2641 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2701 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2761 actcctgatg atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta
     2821 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2881 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgactc
     2941 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3001 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3061 ccggattcag tagtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3121 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3181 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3241 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3301 tttttctgat tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     3361 aataagtgat aataagcgga tgaatggcag aaattcgatg ataagctgtc aaacatgatg
     3421 ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3481 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3541 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3601 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3661 gccagcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3721 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3781 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3841 taacgcgaat tttaacaaaa tattaacgct tacaattt
//
