LOCUS       sequence-231499-        8058 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 8058)
  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 8058)
  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-KIAA2013-201
FEATURES             Location/Qualifiers
     source          1..8058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     343..362
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      451..460
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             457..2109
                     /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    2985..3106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2996..3015)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3050..3069
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3351..3708
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3351..3542
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3374..3394)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3559..3694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3753..3762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3759..4691
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4743..5534
                     /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(4794..4813)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5404..5423
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5598..5646
                     /note="synthetic polyadenylation signal"
     promoter        5802..5906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5907..5975,5976..6767)
                     /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(6125..6144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6925..7513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7005..7024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7629..7912
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(7969..8058,1..410)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8004..8027)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
       61 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
      121 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
      181 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
      241 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
      301 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
      361 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
      421 caagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa aaacattaag
      481 aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca gctgcacaaa
      541 gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc acatatcgag
      601 gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga agctatgaag
      661 cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag cttgcagttc
      721 ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc taacgacatc
      781 tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg
      841 agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat catacaaaag
      901 atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta caccttcgtg
      961 acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag cttcgaccgg
     1021 gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc caagggcgta
     1081 gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc
     1141 aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca cggcttcggc
     1201 atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc
     1261 gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc cctgctggtg
     1321 cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga cctaagcaac
     1381 ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc
     1441 aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac aaccagcgcc
     1501 attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt ggtgcccttc
     1561 ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc
     1621 gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc cgaggctaca
     1681 aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta ctgggacgag
     1741 gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa gggctaccag
     1801 gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt cgacgccggg
     1861 gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa
     1921 cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca ggttacaacc
     1981 gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg actgaccggc
     2041 aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc
     2101 gccgtgtaat tctagttgtt taaacgagct cggtgaacta acagtcctgc tttcagccac
     2161 catttgcaca agacacccag cactgaaagt cccgctgcca ggagcaaggg atcctttgga
     2221 agcacccgcc ctttgtgcct tgttggggga aaccggtgac gcagaagtga gtgtggatac
     2281 accagagttt gcattggaag gaatgagtgt cacgtgggga gggaaggggc cagtggacct
     2341 tttgtaagct ttccactcaa taaaatgaac ctgtatggca aatacttgaa atggaactca
     2401 ctccttccac tttccccctt tcttctgtcc caggaaatag atcatctttt gaaaagactc
     2461 ttgtctagga aaagttgtgt ccttttccta atttaacgtg ttctttctta atgaagtttt
     2521 aatttatttt tgttgagatt ttgctagatg gcttttgcat cccctgtaga tggtgagtgt
     2581 tggcggtgat gtccatctcg gcgttcggag gccccacggt cccgaggctg ggccggggcc
     2641 ccccagggtg gctgtgctgc tgcctgtagg agggtgcggg ttgtgctgtc atcctcgggt
     2701 ttgcacgccc ttttttagga gcctgtggac atctgtggtt ttgtactttg gggcttcagg
     2761 ggaggtgttt aactttctag tgattgatga ttgtcaggtt ttgaaatacc aaagcttttt
     2821 tgttctgttt ttaaataaat atctttcaaa ctttcaagtc gacctgcagg catgcaagct
     2881 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2941 taactagcat aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga
     3001 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3061 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3121 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     3181 cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat
     3241 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     3301 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     3361 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3541 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3601 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3661 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct
     3721 tctgacacaa cagtctcgaa ccaaaggctg gagccaccat ggcttccaag gtgtacgacc
     3781 ccgagcaacg caaacgcatg atcactgggc ctcagtggtg ggctcgctgc aagcaaatga
     3841 acgtgctgga ctccttcatc aactactatg attccgagaa gcacgccgag aacgccgtga
     3901 tttttctgca tggtaacgct gcctccagct acctgtggag gcacgtcgtg cctcacatcg
     3961 agcccgtggc tagatgcatc atccctgatc tgatcggaat gggtaagtcc ggcaagagcg
     4021 ggaatggctc atatcgcctc ctggatcact acaagtacct caccgcttgg ttcgagctgc
     4081 tgaaccttcc aaagaaaatc atctttgtgg gccacgactg gggggcttgt ctggcctttc
     4141 actactccta cgagcaccaa gacaagatca aggccatcgt ccatgctgag agtgtcgtgg
     4201 acgtgatcga gtcctgggac gagtggcctg acatcgagga ggatatcgcc ctgatcaaga
     4261 gcgaagaggg cgagaaaatg gtgcttgaga ataacttctt cgtcgagacc atgctcccaa
     4321 gcaagatcat gcggaaactg gagcctgagg agttcgctgc ctacctggag ccattcaagg
     4381 agaagggcga ggttagacgg cctaccctct cctggcctcg cgagatccct ctcgttaagg
     4441 gaggcaagcc cgacgtcgtc cagattgtcc gcaactacaa cgcctacctt cgggccagcg
     4501 acgatctgcc taagatgttc atcgagtccg accctgggtt cttttccaac gctattgtcg
     4561 agggagctaa gaagttccct aacaccgagt tcgtgaaggt gaagggcctc cacttcagcc
     4621 aggaggacgc tccagatgaa atgggtaagt acatcaagag cttcgtggag cgcgtgctga
     4681 agaacgagca gaccggtggt gggagcggag gtggcggatc aggtggcgga ggctccggag
     4741 ggattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4801 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4861 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4921 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4981 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5041 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5101 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5161 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5221 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5281 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5341 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5401 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5461 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5521 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5581 gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     5641 tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc gttttccctt gtccagatag
     5701 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtaa
     5761 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5821 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5881 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5941 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6001 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6061 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc actttcaaag
     6121 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6181 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6241 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6301 cggccaactt acttctgaca actatcggag gaccgaagga gctaaccgct tttttgcaca
     6361 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6421 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6481 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6541 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6601 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6661 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6721 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaattc gaaatgaccg
     6781 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6841 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6901 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6961 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7021 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7081 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7141 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7201 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7261 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7321 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7381 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7441 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7501 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7561 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7621 tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc
     7681 atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg
     7741 cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt
     7801 tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta
     7861 caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc
     7921 gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatctaccgg gtaggggagg
     7981 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
     8041 acacaagtgg cctctggc
//
