LOCUS       sequence-231691-        7812 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 7812)
  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 7812)
  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-SERINC2-202
FEATURES             Location/Qualifiers
     source          1..7812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..41)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     415..434
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      523..532
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             529..2181
                     /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    2811..2932
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2822..2841)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2876..2895
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3177..3534
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3177..3368
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3200..3220)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3385..3520
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3579..3588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3585..4517
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4569..5360
                     /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(4620..4639)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5230..5249
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5424..5472
                     /note="synthetic polyadenylation signal"
     promoter        5628..5732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5733..5801,5802..6593)
                     /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(5951..5970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6751..7339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6831..6850)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7455..7738
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(7795..7812,1..482)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
ORIGIN
        1 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
       61 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
      121 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
      181 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
      241 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
      301 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
      361 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
      421 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
      481 cgacctgcag cccaagcttg gcaatccggt actgttggta aagccaccat ggaagatgcc
      541 aaaaacatta agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag
      601 cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac
      661 gcacatatcg aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca
      721 gaagctatga agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat
      781 agcttgcagt tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca
      841 gctaacgaca tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc
      901 gtcgtattcg tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg
      961 atcatacaaa agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg
     1021 tacaccttcg tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag
     1081 agcttcgacc gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg
     1141 cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac
     1201 cccatcttcg gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac
     1261 cacggcttcg gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc
     1321 atgtaccgct tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct
     1381 gccctgctgg tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac
     1441 gacctaagca acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt
     1501 gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa
     1561 acaaccagcg ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag
     1621 gtggtgccct tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg
     1681 aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac
     1741 cccgaggcta caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc
     1801 tactgggacg aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac
     1861 aagggctacc aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc
     1921 ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc
     1981 gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc
     2041 caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa
     2101 ggactgaccg gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag
     2161 ggcggcaaga tcgccgtgta attctagttg tttaaacgag ctcgggcagc ctcacagcct
     2221 gccatctggt gcctcctgcc acctggtgcc tctcggctcg gtgacagcca acctgccccc
     2281 tccccacacc aatcagccag gctgagcccc cacccctgcc ccagctccag gacctgcccc
     2341 tgagccgggc cttctagtcg tagtgccttc agggtccgag gagcatcagg ctcctgcaga
     2401 gccccatccc cccgccacac ccacacggtg gagctgcctc ttccttcccc tcctccctgt
     2461 tgcccatact cagcatctcg gatgaaaggg ctcccttgtc ctcaggctcc acgggagcgg
     2521 ggctgctgga gagagcgggg aactcccacc acagtggggc atccggcact gaagccctgg
     2581 tgttcctggt cacgtccccc aggggaccct gcccccttcc tggacttcgt gccttactga
     2641 gtctctaaga ctttttctaa taaacaagcc agtgcgtgta ccagtcgacc tgcaggcatg
     2701 caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2761 gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga taagatacat
     2821 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2881 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2941 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     3001 gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg cttacaattt
     3061 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc
     3121 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     3181 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3421 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3481 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     3541 gattcttctg acacaacagt ctcgaaccaa aggctggagc caccatggct tccaaggtgt
     3601 acgaccccga gcaacgcaaa cgcatgatca ctgggcctca gtggtgggct cgctgcaagc
     3661 aaatgaacgt gctggactcc ttcatcaact actatgattc cgagaagcac gccgagaacg
     3721 ccgtgatttt tctgcatggt aacgctgcct ccagctacct gtggaggcac gtcgtgcctc
     3781 acatcgagcc cgtggctaga tgcatcatcc ctgatctgat cggaatgggt aagtccggca
     3841 agagcgggaa tggctcatat cgcctcctgg atcactacaa gtacctcacc gcttggttcg
     3901 agctgctgaa ccttccaaag aaaatcatct ttgtgggcca cgactggggg gcttgtctgg
     3961 cctttcacta ctcctacgag caccaagaca agatcaaggc catcgtccat gctgagagtg
     4021 tcgtggacgt gatcgagtcc tgggacgagt ggcctgacat cgaggaggat atcgccctga
     4081 tcaagagcga agagggcgag aaaatggtgc ttgagaataa cttcttcgtc gagaccatgc
     4141 tcccaagcaa gatcatgcgg aaactggagc ctgaggagtt cgctgcctac ctggagccat
     4201 tcaaggagaa gggcgaggtt agacggccta ccctctcctg gcctcgcgag atccctctcg
     4261 ttaagggagg caagcccgac gtcgtccaga ttgtccgcaa ctacaacgcc taccttcggg
     4321 ccagcgacga tctgcctaag atgttcatcg agtccgaccc tgggttcttt tccaacgcta
     4381 ttgtcgaggg agctaagaag ttccctaaca ccgagttcgt gaaggtgaag ggcctccact
     4441 tcagccagga ggacgctcca gatgaaatgg gtaagtacat caagagcttc gtggagcgcg
     4501 tgctgaagaa cgagcagacc ggtggtggga gcggaggtgg cggatcaggt ggcggaggct
     4561 ccggagggat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4621 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4681 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4741 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4801 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4861 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4921 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4981 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5041 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5101 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5161 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5221 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5281 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5341 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5401 caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     5461 gttttttgtg tgaatcgata gcgataagga tcctctttgc gcttgcgttt tcccttgtcc
     5521 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     5581 acgtaatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5701 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5761 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5821 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5881 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5941 tcaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6001 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6061 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6121 acactgcggc caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt
     6181 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6241 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6301 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6361 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6421 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6481 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6541 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa
     6601 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     6661 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6721 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6781 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6841 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6901 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6961 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7021 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7081 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7141 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7201 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7261 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7321 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7381 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7441 gatcctttta tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt
     7501 gaaaccatga aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc
     7561 tggttgcata ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg
     7621 gtggtttgtt gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg
     7681 tgcgtacaat taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga
     7741 taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc taccgggtag
     7801 gggaggcgct tt
//
