LOCUS       sequence-233126-        7639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual luciferase assay for miRNA-targeted UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7639)
  AUTHORS   Kutsche LK, Gysi DM, Fallmann J, Lenk K, Petri R, Swiersy A, Klapper
            SD, Pircs K, Khattak S, Stadler PF, Jakobsson J, Nowick K, Busskamp 
            V
  TITLE     Combined Experimental and System-Level Analyses Reveal the Complex 
            Regulatory Network of miR-124 during Human Neurogenesis.
  JOURNAL   Cell Syst. 2018 Sep 28. pii: S2405-4712(18)30358-2. doi: 
            10.1016/j.cels.2018.08.011.
  PUBMED    30292704
REFERENCE   2  (bases 1 to 7639)
  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: "Cell Syst. 
            2018 Sep 28. pii: S2405-4712(18)30358-2. doi: 
            10.1016/j.cels.2018.08.011."
COMMENT     Sequence Label: pmiRGLO-RPAIN-202
FEATURES             Location/Qualifiers
     source          1..7639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        43..542
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(78..101)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     475..494
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      583..592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             589..2241
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2698..2819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2709..2728)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2763..2782
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3064..3421
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3064..3255
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3087..3107)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3272..3407
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3466..3475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3472..4404
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4456..5247
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4507..4526)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5117..5136
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5311..5359
                     /note="synthetic polyadenylation signal"
     promoter        5515..5619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5620..5688,5689..6480)
                     /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(5838..5857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6638..7226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6718..6737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7342..7625
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ctccggccac gatgcgtccg gcgtagagga tcgagatcta ccgggtaggg gaggcgcttt
       61 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
      121 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
      181 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
      241 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
      301 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
      361 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
      421 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
      481 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
      541 cgacctgcag cccaagcttg gcaatccggt actgttggta aagccaccat ggaagatgcc
      601 aaaaacatta agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag
      661 cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac
      721 gcacatatcg aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca
      781 gaagctatga agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat
      841 agcttgcagt tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca
      901 gctaacgaca tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc
      961 gtcgtattcg tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg
     1021 atcatacaaa agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg
     1081 tacaccttcg tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag
     1141 agcttcgacc gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg
     1201 cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac
     1261 cccatcttcg gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac
     1321 cacggcttcg gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc
     1381 atgtaccgct tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct
     1441 gccctgctgg tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac
     1501 gacctaagca acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt
     1561 gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa
     1621 acaaccagcg ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag
     1681 gtggtgccct tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg
     1741 aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac
     1801 cccgaggcta caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc
     1861 tactgggacg aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac
     1921 aagggctacc aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc
     1981 ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc
     2041 gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc
     2101 caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa
     2161 ggactgaccg gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag
     2221 ggcggcaaga tcgccgtgta attctagttg tttaaacgag ctcgagccag cttggactca
     2281 catcattcta tggggttgaa gacaactcat tccctctgag gagccttgta catacaagcc
     2341 ttttatttat aacttatttt gtattgaaac ttttaaacaa tactgaagaa aaaaaaactt
     2401 ttccgacatc tgttcttggt cttttgtgac gcaggttgaa gggggaggaa tagaaaaaga
     2461 caaactgcct tggaggagat aaaccaattt tatgtctatc atgttataca aaaatctaga
     2521 aataatagat ttgtacagaa aaaaatgata ataaatgaga acacaaaaca gtcgacctgc
     2581 aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2641 caccgctgag caataactag cataacccct tggggcggcc gcttcgagca gacatgataa
     2701 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2761 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2821 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     2881 taagcaagta aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt
     2941 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3001 gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     3061 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3241 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3301 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3361 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3421 aaagcttgat tcttctgaca caacagtctc gaaccaaagg ctggagccac catggcttcc
     3481 aaggtgtacg accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc
     3541 tgcaagcaaa tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc
     3601 gagaacgccg tgatttttct gcatggtaac gctgcctcca gctacctgtg gaggcacgtc
     3661 gtgcctcaca tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag
     3721 tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct
     3781 tggttcgagc tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct
     3841 tgtctggcct ttcactactc ctacgagcac caagacaaga tcaaggccat cgtccatgct
     3901 gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc
     3961 gccctgatca agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag
     4021 accatgctcc caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg
     4081 gagccattca aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc
     4141 cctctcgtta agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac
     4201 cttcgggcca gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg gttcttttcc
     4261 aacgctattg tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc
     4321 ctccacttca gccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg
     4381 gagcgcgtgc tgaagaacga gcagaccggt ggtgggagcg gaggtggcgg atcaggtggc
     4441 ggaggctccg gagggattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4501 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4561 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4621 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4681 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4741 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4801 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4861 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4921 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4981 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5041 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5101 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5161 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5221 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5281 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     5341 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc
     5401 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     5461 gctttctacg taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5521 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5581 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5641 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5701 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5761 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5821 agcactttca aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5881 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5941 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6001 agtgataaca ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc
     6061 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6121 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6181 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6241 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6301 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6361 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6421 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6481 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     6541 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6601 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6661 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6721 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6781 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6841 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6901 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6961 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7021 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7081 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7141 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7201 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     7261 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7321 tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg
     7381 ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg
     7441 taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt
     7501 ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt
     7561 ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc
     7621 atcgagataa accgcagta
//
