LOCUS       sequence-162050-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual Fluorescent endogenous miRNA sensor. Perfect complementary 
            region of hsa-miR-27a-3p in three copies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  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 5175)
  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 miR-27a-3p 3 perfect copies.
FEATURES             Location/Qualifiers
     source          1..5175
                     /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    4329..4377
                     /note="synthetic polyadenylation signal"
     promoter        4386..5137
                     /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 tcgagcggaa cttagccact
     4261 gtgaaccggg cggaacttag ccactgtgaa ccgggcggaa cttagccact gtgaaggccg
     4321 ctggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgagg
     4381 atctaaatga gtcttcggac ctcgcggggg ccgcttaagc ggtggttagg gtttgtctga
     4441 cgcgggggga gggggaagga acgaaacact ctcattcgga ggcggctcgg ggtttggtct
     4501 tggtggccac gggcacgcag aagagcgccg cgatcctctt aagcaccccc ccgccctccg
     4561 tggaggcggg ggtttggtcg gcgggtggta actggcgggc cgctgactcg ggcgggtcgc
     4621 gcgccccaga gtgtgacctt ttcggtctgc tcgcagaccc ccgggcggcg ccgccgcggc
     4681 ggcgacgggc tcgctgggtc ctaggctcca tggggaccgt atacgtggac aggctctgga
     4741 gcatccgcac gactgcggtg atattaccgg agaccttctg cgggacgagc cgggtcacgc
     4801 ggctgacgcg gagcgtccgt tgggcgacaa acaccaggac ggggcacagg tacactatct
     4861 tgtcacccgg aggcgcgagg gactgcagga gcttcaggga gtggcgcagc tgcttcatcc
     4921 ccgtggcccg ttgctcgcgt ttgctggcgg tgtccccgga agaaatatat ttgcatgtct
     4981 ttagttctat gatgacacaa accccgccca gcgtcttgtc attggcgaat tcgaacacgc
     5041 agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg acgcgtgtgg
     5101 cctcgaacac cgagcgaccc tgcagcgacc cgcttaaaag cttggcattc cggtactgtt
     5161 ggtaaagccc taaga
//
