LOCUS       sequence-231677-        8025 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 8025)
  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 8025)
  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-TSPAN15-201
FEATURES             Location/Qualifiers
     source          1..8025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..536
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(72..95)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     469..488
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             583..2235
                     /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    3078..3199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3089..3108)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3143..3162
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3444..3801
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3444..3635
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3467..3487)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3652..3787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3846..3855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3852..4784
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4836..5627
                     /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(4887..4906)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5497..5516
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5691..5739
                     /note="synthetic polyadenylation signal"
     promoter        5895..5999
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6000..6068,6069..6860)
                     /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(6218..6237)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7018..7606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7098..7117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7722..8005
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ccacgatgcg tccggcgtag aggatcgaga tctaccgggt aggggaggcg cttttcccaa
       61 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
      121 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg
      181 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
      241 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
      301 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
      361 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg
      421 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc
      481 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct
      541 gcagcccaag cttggcaatc cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac
      601 attaagaagg gcccagcgcc attctaccca ctcgaagacg ggaccgccgg cgagcagctg
      661 cacaaagcca tgaagcgcta cgccctggtg cccggcacca tcgcctttac cgacgcacat
      721 atcgaggtgg acattaccta cgccgagtac ttcgagatga gcgttcggct ggcagaagct
      781 atgaagcgct atgggctgaa tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg
      841 cagttcttca tgcccgtgtt gggtgccctg ttcatcggtg tggctgtggc cccagctaac
      901 gacatctaca acgagcgcga gctgctgaac agcatgggca tcagccagcc caccgtcgta
      961 ttcgtgagca agaaagggct gcaaaagatc ctcaacgtgc aaaagaagct accgatcata
     1021 caaaagatca tcatcatgga tagcaagacc gactaccagg gcttccaaag catgtacacc
     1081 ttcgtgactt cccatttgcc acccggcttc aacgagtacg acttcgtgcc cgagagcttc
     1141 gaccgggaca aaaccatcgc cctgatcatg aacagtagtg gcagtaccgg attgcccaag
     1201 ggcgtagccc taccgcaccg caccgcttgt gtccgattca gtcatgcccg cgaccccatc
     1261 ttcggcaacc agatcatccc cgacaccgct atcctcagcg tggtgccatt tcaccacggc
     1321 ttcggcatgt tcaccacgct gggctacttg atctgcggct ttcgggtcgt gctcatgtac
     1381 cgcttcgagg aggagctatt cttgcgcagc ttgcaagact ataagattca atctgccctg
     1441 ctggtgccca cactatttag cttcttcgct aagagcactc tcatcgacaa gtacgaccta
     1501 agcaacttgc acgagatcgc cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc
     1561 gtggccaaac gcttccacct accaggcatc cgccagggct acggcctgac agaaacaacc
     1621 agcgccattc tgatcacccc cgaaggggac gacaagcctg gcgcagtagg caaggtggtg
     1681 cccttcttcg aggctaaggt ggtggacttg gacaccggta agacactggg tgtgaaccag
     1741 cgcggcgagc tgtgcgtccg tggccccatg atcatgagcg gctacgttaa caaccccgag
     1801 gctacaaacg ctctcatcga caaggacggc tggctgcaca gcggcgacat cgcctactgg
     1861 gacgaggacg agcacttctt catcgtggac cggctgaaga gcctgatcaa atacaagggc
     1921 taccaggtag ccccagccga actggagagc atcctgctgc aacaccccaa catcttcgac
     1981 gccggggtcg ccggcctgcc cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg
     2041 ctggaacacg gtaaaaccat gaccgagaag gagatcgtgg actatgtggc cagccaggtt
     2101 acaaccgcca agaagctgcg cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg
     2161 accggcaagt tggacgcccg caagatccgc gagattctca ttaaggccaa gaagggcggc
     2221 aagatcgccg tgtaattcta gttgtttaaa cgagctcggg cccagcctgc catggcagct
     2281 ccaacaagga ccgtctggga tagcacctct cagtcaacat cgtggggctg gacagggctg
     2341 cggcccctct gcccacactc agtactgacc aaagccaggg ctgtgtgtgc ctgtgtgtag
     2401 gtcccacggc ctctgcctcc ccagggagca gagcctgggc ctcccctaag aggctttccc
     2461 cgaggcagct ctggaatctg tgcccacctg gggcctgggg aacaaggccc tcctttctcc
     2521 aggcctgggc tacgggggag ggagagcctg aggctctgct cagggcccat ttcatctctg
     2581 gcagtgcctt ggcggtggta ttcaaggcag ttttgtagca cctgtaattg gggagaggga
     2641 gtgtgcccct cggggcagga gggaagggca tctggggaag ggcaggaggg aagagctgtc
     2701 catgcagcca cgcccatggc caggttggcc tcttctcagc ctcccaggtg ccttgagccc
     2761 tcttgcaagg gcggctgctt ccttgagcct agttttttta cgtgattttt gtaacattca
     2821 tttttttgta cagataacag gagtttctga ctaatcaaag ctggtatttc cccgcatgtc
     2881 ttattcttgc ccttccccca accagtttgt taatcaaaca ataaaaacat gttttttttt
     2941 ttttttttta gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     3001 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcggcc
     3061 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3121 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3181 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3241 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     3301 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3361 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     3421 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3481 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3601 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3661 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3721 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3781 ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaaccaaagg
     3841 ctggagccac catggcttcc aaggtgtacg accccgagca acgcaaacgc atgatcactg
     3901 ggcctcagtg gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc atcaactact
     3961 atgattccga gaagcacgcc gagaacgccg tgatttttct gcatggtaac gctgcctcca
     4021 gctacctgtg gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc atcatccctg
     4081 atctgatcgg aatgggtaag tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc
     4141 actacaagta cctcaccgct tggttcgagc tgctgaacct tccaaagaaa atcatctttg
     4201 tgggccacga ctggggggct tgtctggcct ttcactactc ctacgagcac caagacaaga
     4261 tcaaggccat cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc
     4321 ctgacatcga ggaggatatc gccctgatca agagcgaaga gggcgagaaa atggtgcttg
     4381 agaataactt cttcgtcgag accatgctcc caagcaagat catgcggaaa ctggagcctg
     4441 aggagttcgc tgcctacctg gagccattca aggagaaggg cgaggttaga cggcctaccc
     4501 tctcctggcc tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc gtccagattg
     4561 tccgcaacta caacgcctac cttcgggcca gcgacgatct gcctaagatg ttcatcgagt
     4621 ccgaccctgg gttcttttcc aacgctattg tcgagggagc taagaagttc cctaacaccg
     4681 agttcgtgaa ggtgaagggc ctccacttca gccaggagga cgctccagat gaaatgggta
     4741 agtacatcaa gagcttcgtg gagcgcgtgc tgaagaacga gcagaccggt ggtgggagcg
     4801 gaggtggcgg atcaggtggc ggaggctccg gagggattga acaagatgga ttgcacgcag
     4861 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4921 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4981 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5041 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5101 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5161 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5221 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5281 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5341 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5401 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5461 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5521 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5581 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5641 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat
     5701 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc
     5761 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     5821 agcaccgttt ctgcggactg gctttctacg taatggtttc ttagacgtca ggtggcactt
     5881 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5941 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6001 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6061 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6121 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6181 aagaacgttt tccaatgatg agcactttca aagttctgct atgtggcgcg gtattatccc
     6241 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6301 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6361 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaactatcg
     6421 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6481 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6541 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6601 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6661 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6721 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6781 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6841 cactgattaa gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc
     6901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7441 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     7621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7681 gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca
     7741 cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta
     7801 agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta
     7861 tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc
     7921 gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact
     7981 tactgtctgc cctcgtagcc atcgagataa accgcagtac tccgg
//
