LOCUS       sequence-231449-        8532 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 8532)
  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 8532)
  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-ARF1-214
FEATURES             Location/Qualifiers
     source          1..8532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     333..352
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      441..450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             447..2099
                     /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    3449..3570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3460..3479)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3514..3533
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3815..4172
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3815..4006
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3838..3858)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4023..4158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4217..4226
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4223..5155
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             5207..5998
                     /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(5258..5277)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5868..5887
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6062..6110
                     /note="synthetic polyadenylation signal"
     promoter        6266..6370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6371..6439,6440..7231)
                     /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(6589..6608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7389..7977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7469..7488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8093..8376
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(8433..8532,1..400)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8468..8491)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
       61 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
      121 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca
      181 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg
      241 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg
      301 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc
      361 tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttggc
      421 aatccggtac tgttggtaaa gccaccatgg aagatgccaa aaacattaag aagggcccag
      481 cgccattcta cccactcgaa gacgggaccg ccggcgagca gctgcacaaa gccatgaagc
      541 gctacgccct ggtgcccggc accatcgcct ttaccgacgc acatatcgag gtggacatta
      601 cctacgccga gtacttcgag atgagcgttc ggctggcaga agctatgaag cgctatgggc
      661 tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg
      721 tgttgggtgc cctgttcatc ggtgtggctg tggccccagc taacgacatc tacaacgagc
      781 gcgagctgct gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag
      841 ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat catacaaaag atcatcatca
      901 tggatagcaa gaccgactac cagggcttcc aaagcatgta caccttcgtg acttcccatt
      961 tgccacccgg cttcaacgag tacgacttcg tgcccgagag cttcgaccgg gacaaaacca
     1021 tcgccctgat catgaacagt agtggcagta ccggattgcc caagggcgta gccctaccgc
     1081 accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc aaccagatca
     1141 tccccgacac cgctatcctc agcgtggtgc catttcacca cggcttcggc atgttcacca
     1201 cgctgggcta cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc
     1261 tattcttgcg cagcttgcaa gactataaga ttcaatctgc cctgctggtg cccacactat
     1321 ttagcttctt cgctaagagc actctcatcg acaagtacga cctaagcaac ttgcacgaga
     1381 tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc
     1441 acctaccagg catccgccag ggctacggcc tgacagaaac aaccagcgcc attctgatca
     1501 cccccgaagg ggacgacaag cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta
     1561 aggtggtgga cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg
     1621 tccgtggccc catgatcatg agcggctacg ttaacaaccc cgaggctaca aacgctctca
     1681 tcgacaagga cggctggctg cacagcggcg acatcgccta ctgggacgag gacgagcact
     1741 tcttcatcgt ggaccggctg aagagcctga tcaaatacaa gggctaccag gtagccccag
     1801 ccgaactgga gagcatcctg ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc
     1861 tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa
     1921 ccatgaccga gaaggagatc gtggactatg tggccagcca ggttacaacc gccaagaagc
     1981 tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg actgaccggc aagttggacg
     2041 cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc gccgtgtaat
     2101 tctagttgtt taaacgagct cgacgcgacc cccctccctc tcactcctct tgccctctgc
     2161 tttactctca tgtggcaaac gtgcggctcg tggtgtgagt gccagaagct gcctccgtgg
     2221 tttggtcacc gtgtgcatcg caccgtgctg taaatgtggc agacgcagcc tgcggccagg
     2281 ctttttattt aatgtaaata gtttttgttt ccaatgaggc agtttctggt actcctatgc
     2341 aatattactc agcttttttt attgtaaaaa gaaaaatcaa ctcactgttc agtgctgaga
     2401 ggggatgtag gcccatgggc acctggcctc caggagtcgc tgtgttggga gagccggcca
     2461 cgcccttggc tttagagctg tgttgaaatc cattttggtg gttggttttt aacccaaact
     2521 cagtgcattt tttaaaatag ttaagaatcc aagtcgagaa cacttgaaca cacagaaggg
     2581 agaccccgcc tagcatagat ttgcagttac ggcctggatg ccagtcgcca gcccagctgt
     2641 tcccctcggg aacatgaggt ggtggtggcg cagcagactg cgatcaattc tgcatggtca
     2701 cagtagagat ccccgcaact cgcttgtcct tgggtcaccc tgcattccat agccatgtgc
     2761 ttgtccctgt gctcccacgg ttcccagggg ccaggctggg agcccacagc caccccacta
     2821 tgccgcaggc cgccctaccc accttcaggc agcctatggg acgcagggcc ccatctgtcc
     2881 ctcggtcgcc gtgtggccag agtgggtccg tcgtccccaa cactcgtgct cgctcagaca
     2941 ctttggcagg atgtctgggg cctcaccagc aggagcgcgt gcaagccggg caggcggtcc
     3001 acctagaccc acagcccctc gggagcaccc cacctctgtg tgtgatgtag ctttctctcc
     3061 ctcagcctgc aagggtccga tttgccatcg aaaaagacaa cctctacttt tttcttttgt
     3121 attttgataa acactgaagc tggagctgtt aaatttatct tggggaaacc tcagaactgg
     3181 tctatttggt gtcgtggaac ctcttactgc tttcaataca cgattagtaa tcaactgttt
     3241 tgtatacttg ttttcagttt tcatttcgac aaacaagcac tgtaattata gctattagaa
     3301 taaaatctct taactatttc agtcgacctg caggcatgca agctgatccg gctgctaaca
     3361 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     3421 ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3481 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3541 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3601 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     3661 aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     3721 acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa
     3781 taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga
     3841 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3901 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3961 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4021 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4081 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4141 gaggcttttt tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct
     4201 cgaaccaaag gctggagcca ccatggcttc caaggtgtac gaccccgagc aacgcaaacg
     4261 catgatcact gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc tggactcctt
     4321 catcaactac tatgattccg agaagcacgc cgagaacgcc gtgatttttc tgcatggtaa
     4381 cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac atcgagcccg tggctagatg
     4441 catcatccct gatctgatcg gaatgggtaa gtccggcaag agcgggaatg gctcatatcg
     4501 cctcctggat cactacaagt acctcaccgc ttggttcgag ctgctgaacc ttccaaagaa
     4561 aatcatcttt gtgggccacg actggggggc ttgtctggcc tttcactact cctacgagca
     4621 ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga tcgagtcctg
     4681 ggacgagtgg cctgacatcg aggaggatat cgccctgatc aagagcgaag agggcgagaa
     4741 aatggtgctt gagaataact tcttcgtcga gaccatgctc ccaagcaaga tcatgcggaa
     4801 actggagcct gaggagttcg ctgcctacct ggagccattc aaggagaagg gcgaggttag
     4861 acggcctacc ctctcctggc ctcgcgagat ccctctcgtt aagggaggca agcccgacgt
     4921 cgtccagatt gtccgcaact acaacgccta ccttcgggcc agcgacgatc tgcctaagat
     4981 gttcatcgag tccgaccctg ggttcttttc caacgctatt gtcgagggag ctaagaagtt
     5041 ccctaacacc gagttcgtga aggtgaaggg cctccacttc agccaggagg acgctccaga
     5101 tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg agcagaccgg
     5161 tggtgggagc ggaggtggcg gatcaggtgg cggaggctcc ggagggattg aacaagatgg
     5221 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5281 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5341 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5401 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5461 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5521 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5581 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5641 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5701 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5761 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5821 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5881 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5941 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6001 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     6061 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     6121 gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     6181 catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt cttagacgtc
     6241 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6301 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6361 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6421 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6481 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6541 ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc tatgtggcgc
     6601 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6661 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6721 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6781 gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6841 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6901 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6961 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     7021 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7081 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7141 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7201 gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag cgacgcccaa
     7261 ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7801 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa
     7981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     8041 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaaat
     8101 acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa aatggcagct
     8161 tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg tattcatacg
     8221 gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc catttttacc
     8281 tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta agggattatg
     8341 gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac
     8401 gatgcgtccg gcgtagagga tcgagatcta ccgggtaggg gaggcgcttt tcccaaggca
     8461 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     8521 gcctcgcaca ca
//
