LOCUS       sequence-165664-        5139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  [Edit] Dual Fluorescent endogenous miRNA sensor. To insert the 
            complementary region of the 3'UTR of RFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5139)
  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 5139)
  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.3.0 Empty
FEATURES             Location/Qualifiers
     source          1..5139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             32..751
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    802..923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(813..832)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     867..886
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1056..1160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1161..1229,1230..2021)
                     /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(1379..1398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2192..2780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2681..2700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2909..3266
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2909..3100
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2932..2952)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3117..3252
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3336..3468
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3451..3470
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        3513..3531
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3553..4248
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     polyA_signal    4294..4342
                     /note="synthetic polyadenylation signal"
     promoter        4351..5102
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
ORIGIN
        1 aacattaaga agggcccggg atccaccggt catggtgagc aagggcgccg agctgttcac
       61 cggcatcgtg cccatcctga tcgagctgaa tggcgatgtg aatggccaca agttcagcgt
      121 gagcggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      181 caccggcaag ctgcctgtgc cctggcccac cctggtgacc accctgagct acggcgtgca
      241 gtgcttctca cgctaccccg atcacatgaa gcagcacgac ttcttcaaga gcgccatgcc
      301 tgagggctac atccaggagc gcaccatctt cttcgaggat gacggcaact acaagtcgcg
      361 cgccgaggtg aagttcgagg gcgataccct ggtgaatcgc atcgagctga ccggcaccga
      421 tttcaaggag gatggcaaca tcctgggcaa taagatggag tacaactaca acgcccacaa
      481 tgtgtacatc atgaccgaca aggccaagaa tggcatcaag gtgaacttca agatccgcca
      541 caacatcgag gatggcagcg tgcagctggc cgaccactac cagcagaata cccccatcgg
      601 cgatggccct gtgctgctgc ccgataacca ctacctgtcc acccagagcg ccctgtccaa
      661 ggaccccaac gagaagcgcg atcacatgat ctacttcggc ttcgtgaccg ccgccgccat
      721 cacccacggc atggatgagc tgtacaagtg agctgccgcg actctagagt cggggcggcc
      781 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
      841 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
      901 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
      961 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     1021 ataaggatcc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1081 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1141 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1201 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1261 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1321 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1381 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1441 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1501 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1561 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1621 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1681 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1741 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1801 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1861 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1921 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1981 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2041 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2101 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2161 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2221 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2281 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2341 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2401 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2461 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2521 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2581 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2641 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2701 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2761 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2821 gctggccttt tgctcacatg gctcgacaga tctgcgcagc accatggcct gaaataacct
     2881 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     2941 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3001 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3061 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3121 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3181 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3241 tttttggagg cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact
     3301 taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg
     3361 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
     3421 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc
     3481 agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag gctagcgcta
     3541 ccggtcgcca ccatgagcga gctgatcaag gagaacatgc acatgaagct gtacatggag
     3601 ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag
     3661 ggcacccaga ccatgaagat caaggtggtc gagggcggcc ctctcccctt cgccttcgac
     3721 atcctggcta ccagcttcat gtacggcagc aaagccttca tcaaccacac ccagggcatc
     3781 cccgacttct ttaagcagtc cttccctgag ggcttcacat gggagagaat caccacatac
     3841 gaagacgggg gcgtgctgac cgctacccag gacaccagct tccagaacgg ctgcatcatc
     3901 tacaacgtca agatcaacgg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa
     3961 acacgcggct gggaggccaa caccgagatg ctgtaccccg ctgacggcgg cctgagaggc
     4021 cacagccaga tggccctgaa gctcgtgggc gggggctacc tgcactgctc cttcaagacc
     4081 acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcttcca cttcgtggac
     4141 cacagactgg aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgagatg
     4201 gctgtggcca agtactgcga cctccctagc aaactggggc acagataact cgagcccggg
     4261 aattcgttta aacctagagc ggccgctggc cgcaataaaa tatctttatt ttcattacat
     4321 ctgtgtgttg gttttttgtg tgaggatcta aatgagtctt cggacctcgc gggggccgct
     4381 taagcggtgg ttagggtttg tctgacgcgg ggggaggggg aaggaacgaa acactctcat
     4441 tcggaggcgg ctcggggttt ggtcttggtg gccacgggca cgcagaagag cgccgcgatc
     4501 ctcttaagca cccccccgcc ctccgtggag gcgggggttt ggtcggcggg tggtaactgg
     4561 cgggccgctg actcgggcgg gtcgcgcgcc ccagagtgtg accttttcgg tctgctcgca
     4621 gacccccggg cggcgccgcc gcggcggcga cgggctcgct gggtcctagg ctccatgggg
     4681 accgtatacg tggacaggct ctggagcatc cgcacgactg cggtgatatt accggagacc
     4741 ttctgcggga cgagccgggt cacgcggctg acgcggagcg tccgttgggc gacaaacacc
     4801 aggacggggc acaggtacac tatcttgtca cccggaggcg cgagggactg caggagcttc
     4861 agggagtggc gcagctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc
     4921 ccggaagaaa tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc
     4981 ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact
     5041 tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt
     5101 aaaagcttgg cattccggta ctgttggtaa agccctaag
//
