LOCUS       sequence-230214-        7521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual luciferase assay positive control for miR-124 binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7521)
  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 7521)
  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-miR-124-sponge (pos. CTRL)
FEATURES             Location/Qualifiers
     source          1..7521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        41..540
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(76..99)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     473..492
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             587..2239
                     /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    2578..2699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2589..2608)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2643..2662
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2944..3301
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2944..3135
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2967..2987)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3152..3287
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3346..3355
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3352..4284
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4336..5127
                     /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(4387..4406)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4997..5016
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5191..5239
                     /note="synthetic polyadenylation signal"
     promoter        5395..5499
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5500..5568,5569..6360)
                     /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(5718..5737)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6518..7106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6598..6617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7222..7505
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ccggccacga tgcgtccggc gtagaggatc gagatctacc gggtagggga ggcgcttttc
       61 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
      121 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
      181 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
      241 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
      301 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
      361 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
      421 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
      481 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
      541 acctgcagcc caagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa
      601 aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca
      661 gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc
      721 acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga
      781 agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag
      841 cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc
      901 taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt
      961 cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat
     1021 catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta
     1081 caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag
     1141 cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc
     1201 caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc
     1261 catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca
     1321 cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat
     1381 gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc
     1441 cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga
     1501 cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga
     1561 ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac
     1621 aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt
     1681 ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa
     1741 ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc
     1801 cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta
     1861 ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa
     1921 gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt
     1981 cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt
     2041 cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca
     2101 ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg
     2161 actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg
     2221 cggcaagatc gccgtgtaat tctagttgtt taaacgagcc gatggaaccg tcactgatat
     2281 gtctaaaggc attcaacggt gccttaaatc ctttctaatc caaaatggca ttcaaaagtg
     2341 ccttaactcc aaaactccaa tcaatggcat tcaaaggtgc cttatttcgt aaataaaatc
     2401 aaatggcatt caaatgtgcc ttagtcgaca gtaggaattc gtcgagggga gtcgacctgc
     2461 aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2521 caccgctgag caataactag cataacccct tggggcggcc gcttcgagca gacatgataa
     2581 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2641 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2701 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     2761 taagcaagta aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt
     2821 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2881 gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     2941 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3001 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3061 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3121 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3181 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3241 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3301 aaagcttgat tcttctgaca caacagtctc gaaccaaagg ctggagccac catggcttcc
     3361 aaggtgtacg accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc
     3421 tgcaagcaaa tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc
     3481 gagaacgccg tgatttttct gcatggtaac gctgcctcca gctacctgtg gaggcacgtc
     3541 gtgcctcaca tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag
     3601 tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct
     3661 tggttcgagc tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct
     3721 tgtctggcct ttcactactc ctacgagcac caagacaaga tcaaggccat cgtccatgct
     3781 gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc
     3841 gccctgatca agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag
     3901 accatgctcc caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg
     3961 gagccattca aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc
     4021 cctctcgtta agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac
     4081 cttcgggcca gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg gttcttttcc
     4141 aacgctattg tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc
     4201 ctccacttca gccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg
     4261 gagcgcgtgc tgaagaacga gcagaccggt ggtgggagcg gaggtggcgg atcaggtggc
     4321 ggaggctccg gagggattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4381 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4441 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4501 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4561 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4621 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4681 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4741 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4801 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4861 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4921 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4981 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5041 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5101 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5161 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     5221 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc
     5281 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     5341 gctttctacg taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5401 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5461 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5521 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5581 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5641 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5701 agcactttca aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5761 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5821 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5881 agtgataaca ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc
     5941 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6001 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6061 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6121 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6181 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6241 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6301 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6361 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     6421 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6481 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6541 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6601 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6661 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6721 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6781 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6841 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6901 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6961 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7021 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7081 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     7141 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7201 tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg
     7261 ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg
     7321 taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt
     7381 ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt
     7441 ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc
     7501 atcgagataa accgcagtac t
//
