LOCUS       sequence-165424-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  [Edit] Dual Fluorescent endogenous miRNA sensor. To insert the 
            complementary region at the 3'UTR of CFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  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 5131)
  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 Empty
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(18..20,21..23,24..737)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    777..898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(788..807)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     842..861
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1031..1135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1136..1204,1205..1996)
                     /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(1354..1373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2167..2755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2656..2675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2884..3241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2884..3075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2907..2927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3092..3227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3311..3443
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3426..3445
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        3488..3506
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3514..3516,3517..3519,3520..4233)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4286..4334
                     /note="synthetic polyadenylation signal"
     promoter        4343..5094
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
ORIGIN
        1 aacattaaga agggcccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
       61 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      121 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      181 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct
      241 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      301 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      361 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
      421 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
      481 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
      541 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
      601 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
      661 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
      721 acgagctgta caagtgatct agagtcgggg cggccggccg cttcgagcag acatgataag
      781 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      841 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      901 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      961 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccaggtg gcacttttcg
     1021 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1081 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1141 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1201 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1261 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1321 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1381 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1441 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1501 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1561 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1621 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1681 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1741 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1801 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1861 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1921 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1981 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2041 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2101 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2161 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2221 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2281 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2341 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2401 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2461 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2521 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2581 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2641 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2701 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2761 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     2821 acagatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     2881 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2941 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3001 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3061 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3121 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3181 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3241 aaagcttgat tcttctgaca caacagtctc gaacttaagc tgcagaagtt ggtcgtgagg
     3301 cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     3361 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
     3421 ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct cttaaggcta
     3481 gagtacttaa tacgactcac tataggctag ccaatggtga gcaagggcga ggagctgttc
     3541 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caggttcagc
     3601 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     3661 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg
     3721 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3781 cccgaaggct acgtccagga gcgtaccatc ttcttcaagg acgacggcaa ctacaagacc
     3841 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3901 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac
     3961 aacgtctata tcaccgccga caagcagaag aacggcatca aggcccactt caagatccgc
     4021 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     4081 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     4141 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     4201 atcactctcg gcatggacga gctgtacaag taagcgatcg ctcgagcccg ggaattcgtt
     4261 taaacctaga gcggccgctg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4321 tggttttttg tgtgaggatc taaatgagtc ttcggacctc gcgggggccg cttaagcggt
     4381 ggttagggtt tgtctgacgc ggggggaggg ggaaggaacg aaacactctc attcggaggc
     4441 ggctcggggt ttggtcttgg tggccacggg cacgcagaag agcgccgcga tcctcttaag
     4501 cacccccccg ccctccgtgg aggcgggggt ttggtcggcg ggtggtaact ggcgggccgc
     4561 tgactcgggc gggtcgcgcg ccccagagtg tgaccttttc ggtctgctcg cagacccccg
     4621 ggcggcgccg ccgcggcggc gacgggctcg ctgggtccta ggctccatgg ggaccgtata
     4681 cgtggacagg ctctggagca tccgcacgac tgcggtgata ttaccggaga ccttctgcgg
     4741 gacgagccgg gtcacgcggc tgacgcggag cgtccgttgg gcgacaaaca ccaggacggg
     4801 gcacaggtac actatcttgt cacccggagg cgcgagggac tgcaggagct tcagggagtg
     4861 gcgcagctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga
     4921 aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt
     4981 ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat
     5041 taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaaaagctt
     5101 ggcattccgg tactgttggt aaagccctaa g
//
