LOCUS       sequence-231451-        8020 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 8020)
  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 8020)
  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-RPLP1-201
FEATURES             Location/Qualifiers
     source          1..8020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..519
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(55..78)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     452..471
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      560..569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             566..2218
                     /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    3056..3177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3067..3086)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3121..3140
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3422..3779
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3422..3613
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3445..3465)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3630..3765
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3824..3833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3830..4762
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4814..5605
                     /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(4865..4884)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5475..5494
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5669..5717
                     /note="synthetic polyadenylation signal"
     promoter        5873..5977
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5978..6046,6047..6838)
                     /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(6196..6215)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6996..7584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7076..7095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7700..7983
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tagaggatcg agatctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
       61 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
      121 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
      181 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
      241 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
      301 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
      361 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
      421 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
      481 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttggca
      541 atccggtact gttggtaaag ccaccatgga agatgccaaa aacattaaga agggcccagc
      601 gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg
      661 ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac
      721 ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct
      781 gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt
      841 gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg
      901 cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg
      961 gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat
     1021 ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt
     1081 gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat
     1141 cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca
     1201 ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat
     1261 ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac
     1321 gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct
     1381 attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt
     1441 tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat
     1501 cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca
     1561 cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac
     1621 ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa
     1681 ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt
     1741 ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat
     1801 cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt
     1861 cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc
     1921 cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct
     1981 gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac
     2041 catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct
     2101 gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc
     2161 ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgtaatt
     2221 ctagttgttt aaacgagctc gacctctttt ataacatgtt caataaaaag ctgaacttta
     2281 ctgctgttgg tcttgtccat agttttggaa tgtgctctgc aaaaatggtc tgttttgtaa
     2341 tgttggcttt cagcctattc tgccatgaca caggctggat tttccctgcc accattgccg
     2401 gatgtgagat ttagacaatc ctgatgctaa caagaaggca cctcatggta cagtgttgaa
     2461 aactgcactg gggtggcagg aggaaggatc agagcagatg ctttgtccag gttacctgtg
     2521 taaacttgat gattataagg tgtgtcccct acactggggg tggagggtaa tacagaccag
     2581 ctttagaaag cctcctgggt atttacccta tacaaccctg cctaagaacg aactttgacg
     2641 tatgctgaat gttgactgct acatcccaag gttgtaaaaa aacaagactt agctatgtaa
     2701 tgtctcacca aatgtttggg agttttatga taacactgaa ggagtgcaga aagtttactt
     2761 aatagtttat taaggtctcc agttacacaa agtggaaaaa aaaaagattc aaacctgtgt
     2821 tccaaaaact aagagctttt tttgtttgcc aattaataat accatgaatg gaattttcaa
     2881 cttttcaggc ttggatttat ataataaacc gtgggtattg tgtgttcagt cgacctgcag
     2941 gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     3001 ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga
     3061 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3121 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3181 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta
     3241 agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat taacgcttac
     3301 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc
     3361 ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
     3421 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3481 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3541 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3601 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3661 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3721 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3781 agcttgattc ttctgacaca acagtctcga accaaaggct ggagccacca tggcttccaa
     3841 ggtgtacgac cccgagcaac gcaaacgcat gatcactggg cctcagtggt gggctcgctg
     3901 caagcaaatg aacgtgctgg actccttcat caactactat gattccgaga agcacgccga
     3961 gaacgccgtg atttttctgc atggtaacgc tgcctccagc tacctgtgga ggcacgtcgt
     4021 gcctcacatc gagcccgtgg ctagatgcat catccctgat ctgatcggaa tgggtaagtc
     4081 cggcaagagc gggaatggct catatcgcct cctggatcac tacaagtacc tcaccgcttg
     4141 gttcgagctg ctgaaccttc caaagaaaat catctttgtg ggccacgact ggggggcttg
     4201 tctggccttt cactactcct acgagcacca agacaagatc aaggccatcg tccatgctga
     4261 gagtgtcgtg gacgtgatcg agtcctggga cgagtggcct gacatcgagg aggatatcgc
     4321 cctgatcaag agcgaagagg gcgagaaaat ggtgcttgag aataacttct tcgtcgagac
     4381 catgctccca agcaagatca tgcggaaact ggagcctgag gagttcgctg cctacctgga
     4441 gccattcaag gagaagggcg aggttagacg gcctaccctc tcctggcctc gcgagatccc
     4501 tctcgttaag ggaggcaagc ccgacgtcgt ccagattgtc cgcaactaca acgcctacct
     4561 tcgggccagc gacgatctgc ctaagatgtt catcgagtcc gaccctgggt tcttttccaa
     4621 cgctattgtc gagggagcta agaagttccc taacaccgag ttcgtgaagg tgaagggcct
     4681 ccacttcagc caggaggacg ctccagatga aatgggtaag tacatcaaga gcttcgtgga
     4741 gcgcgtgctg aagaacgagc agaccggtgg tgggagcgga ggtggcggat caggtggcgg
     4801 aggctccgga gggattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4861 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4921 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4981 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5041 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5101 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5161 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5221 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5281 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5341 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5401 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5461 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5521 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5581 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5641 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5701 tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct
     5761 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     5821 tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5881 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5941 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6001 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6061 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6121 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6181 cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6241 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6301 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6361 tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg agctaaccgc
     6421 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6481 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6541 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6601 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6661 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6721 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6781 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatt
     6841 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     6901 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6961 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7021 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7081 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7141 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7201 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7261 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7321 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7381 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7441 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7501 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7561 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     7621 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7681 acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc cttcgctggg
     7741 atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat gtggcctgta
     7801 ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt gcggcagttt
     7861 ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt
     7921 agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat
     7981 cgagataaac cgcagtactc cggccacgat gcgtccggcg
//
