LOCUS       sequence-168133-        5127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual Fluorescent endogenous miRNA sensor. Perfect complementary 
            region of an siRNA control targeting firefly luciferase encoded by 
            the pGL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5127)
  AUTHORS   Lemus-Diaz N, Boker KO, Rodriguez-Polo I, Mitter M, Preis J, Arlt M,
            Gruber J
  TITLE     Dissecting miRNA gene repression on single cell level with an 
            advanced fluorescent reporter system.
  JOURNAL   Sci Rep. 2017 Mar 24;7:45197. doi: 10.1038/srep45197.
  PUBMED    28338079
REFERENCE   2  (bases 1 to 5127)
  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: "Sci Rep."; 
            date: "2017-03-24"; volume: "7:45197. doi"; pages: " 
            10.1038/srep45197"
COMMENT     Sequence Label: p.UTA.2.0-GL-2
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(17..19,20..22,23..736)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    776..897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(787..806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     841..860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1030..1134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1135..1203,1204..1995)
                     /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(1353..1372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2166..2754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2655..2674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2883..3240
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2883..3074
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2906..2926)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3091..3226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3310..3442
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3425..3444
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        3487..3505
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3513..3515,3516..3518,3519..4232)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4281..4329
                     /note="synthetic polyadenylation signal"
     promoter        4338..5089
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
ORIGIN
        1 acattaagaa gggcccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
       61 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      121 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      181 cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct tcgcccgcta
      241 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      301 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      361 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      421 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
      481 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
      541 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      601 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
      661 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      721 cgagctgtac aagtgatcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
      781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
      841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
      901 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
      961 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccaggtgg cacttttcgg
     1021 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1081 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1141 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1201 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1261 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1321 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1381 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1441 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1501 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1561 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1621 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1681 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1741 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1801 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1861 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1921 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1981 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2041 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2101 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2161 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2221 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2281 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2341 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2401 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2461 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2521 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2581 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2641 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2701 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2761 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
     2821 cagatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     2881 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3061 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3241 aagcttgatt cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc
     3301 actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
     3361 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     3421 cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag
     3481 agtacttaat acgactcact ataggctagc caatggtgag caagggcgag gagctgttca
     3541 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aggttcagcg
     3601 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     3661 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc
     3721 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     3781 ccgaaggcta cgtccaggag cgtaccatct tcttcaagga cgacggcaac tacaagaccc
     3841 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3901 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac atcagccaca
     3961 acgtctatat caccgccgac aagcagaaga acggcatcaa ggcccacttc aagatccgcc
     4021 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     4081 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     4141 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     4201 tcactctcgg catggacgag ctgtacaagt aagcgatcgc tcgatttcga agtattccgc
     4261 gtacgtgggc cgctggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     4321 ttttgtgtga ggatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta
     4381 gggtttgtct gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc
     4441 ggggtttggt cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc
     4501 ccccgccctc cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact
     4561 cgggcgggtc gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg
     4621 cgccgccgcg gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg
     4681 acaggctctg gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga
     4741 gccgggtcac gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca
     4801 ggtacactat cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca
     4861 gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat
     4921 atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga
     4981 attcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg
     5041 tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttggcat
     5101 tccggtactg ttggtaaagc cctaaga
//
