LOCUS       sequence-231488-        7687 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 7687)
  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 7687)
  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-ARHGAP4-203
FEATURES             Location/Qualifiers
     source          1..7687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        46..545
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(81..104)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     478..497
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      586..595
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             592..2244
                     /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    2749..2870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2760..2779)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2814..2833
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3115..3472
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3115..3306
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3138..3158)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3323..3458
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3517..3526
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3523..4455
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4507..5298
                     /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(4558..4577)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5168..5187
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5362..5410
                     /note="synthetic polyadenylation signal"
     promoter        5566..5670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5671..5739,5740..6531)
                     /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(5889..5908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6689..7277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6769..6788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7393..7676
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ctaccgggta ggggaggcgc
       61 ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca
      121 caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt
      181 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg
      241 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg
      301 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat
      361 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg
      421 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
      481 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc
      541 tttcgacctg cagcccaagc ttggcaatcc ggtactgttg gtaaagccac catggaagat
      601 gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg gaccgccggc
      661 gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat cgcctttacc
      721 gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag cgttcggctg
      781 gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag
      841 aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc
      901 ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat cagccagccc
      961 accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca aaagaagcta
     1021 ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg cttccaaagc
     1081 atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga cttcgtgccc
     1141 gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg cagtaccgga
     1201 ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag tcatgcccgc
     1261 gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt ggtgccattt
     1321 caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt tcgggtcgtg
     1381 ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta taagattcaa
     1441 tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct catcgacaag
     1501 tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag caaggaggta
     1561 ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta cggcctgaca
     1621 gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg cgcagtaggc
     1681 aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa gacactgggt
     1741 gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac
     1801 aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag cggcgacatc
     1861 gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag cctgatcaaa
     1921 tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca acaccccaac
     1981 atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct gcccgccgca
     2041 gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga ctatgtggcc
     2101 agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct
     2161 aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat taaggccaag
     2221 aagggcggca agatcgccgt gtaattctag ttgtttaaac gagctcgggt gccgctgctg
     2281 gagatgcgtg cccccggcgg ctacccgctg gaccggccac tctccccagc ccccttgctt
     2341 ctctccagcc ctgtccagca agtgcagggt gcctgcactt caccctgtgc agagaggtgg
     2401 gatggggccg tgcacacagg gatgcccgct ccacatcctg cctgcccctc agccctggcc
     2461 caggcccctt ttggaggcag ctgaggaagg atgctgggga aagccctctt ctgcagcttt
     2521 gtggaaggct gatcagtggc tgctgggtgg cgggtaccct tgctcagatg cctggcaggg
     2581 ctgggtggcg attcataaag acctcgtgtt gattccccga agtcgacctg caggcatgca
     2641 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     2701 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     2761 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2821 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2881 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     2941 aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc
     3001 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     3061 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     3121 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3181 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3241 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3301 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3361 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3421 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga
     3481 ttcttctgac acaacagtct cgaaccaaag gctggagcca ccatggcttc caaggtgtac
     3541 gaccccgagc aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa
     3601 atgaacgtgc tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc
     3661 gtgatttttc tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac
     3721 atcgagcccg tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag
     3781 agcgggaatg gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag
     3841 ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc
     3901 tttcactact cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc
     3961 gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc
     4021 aagagcgaag agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc
     4081 ccaagcaaga tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc
     4141 aaggagaagg gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt
     4201 aagggaggca agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc
     4261 agcgacgatc tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt
     4321 gtcgagggag ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc
     4381 agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg
     4441 ctgaagaacg agcagaccgg tggtgggagc ggaggtggcg gatcaggtgg cggaggctcc
     4501 ggagggattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4561 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4621 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4681 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4741 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4801 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4861 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4921 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4981 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5041 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5101 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5161 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5221 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5281 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5341 acctgccatc acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt
     5401 tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc ccttgtccag
     5461 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     5521 gtaatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5641 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5701 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5761 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5821 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttc
     5881 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5941 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6001 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6061 actgcggcca acttacttct gacaactatc ggaggaccga aggagctaac cgcttttttg
     6121 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6181 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6241 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6301 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6361 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6421 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6481 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta attcgaaatg
     6541 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     6601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7261 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7381 tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga
     7441 aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg
     7501 gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt
     7561 ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg
     7621 cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata
     7681 aaccgca
//
