LOCUS       sequence-231687-        7610 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 7610)
  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 7610)
  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-SRM-201
FEATURES             Location/Qualifiers
     source          1..7610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..554
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(90..113)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     487..506
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             601..2253
                     /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    2681..2802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2692..2711)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2746..2765
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3047..3404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3047..3238
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3070..3090)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3255..3390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3449..3458
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3455..4387
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4439..5230
                     /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(4490..4509)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5100..5119
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5294..5342
                     /note="synthetic polyadenylation signal"
     promoter        5498..5602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5603..5671,5672..6463)
                     /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(5821..5840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6621..7209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6701..6720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7325..7608
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc taccgggtag
       61 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
      121 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
      181 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
      241 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
      301 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
      361 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
      421 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
      481 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
      541 ctccgggcct ttcgacctgc agcccaagct tggcaatccg gtactgttgg taaagccacc
      601 atggaagatg ccaaaaacat taagaagggc ccagcgccat tctacccact cgaagacggg
      661 accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc cggcaccatc
      721 gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt cgagatgagc
      781 gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg gatcgtggtg
      841 tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg
      901 gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag catgggcatc
      961 agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa
     1021 aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga ctaccagggc
     1081 ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa cgagtacgac
     1141 ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa cagtagtggc
     1201 agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt
     1261 catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat cctcagcgtg
     1321 gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat ctgcggcttt
     1381 cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt gcaagactat
     1441 aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa gagcactctc
     1501 atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc gccgctcagc
     1561 aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg ccagggctac
     1621 ggcctgacag aaacaaccag cgccattctg atcacccccg aaggggacga caagcctggc
     1681 gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga caccggtaag
     1741 acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat catgagcggc
     1801 tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg gctgcacagc
     1861 ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg gctgaagagc
     1921 ctgatcaaat acaagggcta ccaggtagcc ccagccgaac tggagagcat cctgctgcaa
     1981 caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg
     2041 cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga gatcgtggac
     2101 tatgtggcca gccaggttac aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac
     2161 gaggtgccta aaggactgac cggcaagttg gacgcccgca agatccgcga gattctcatt
     2221 aaggccaaga agggcggcaa gatcgccgtg taattctagt tgtttaaacg agctcggccc
     2281 aggcgccacc actgatgcca cccaggacct cggaccttgg agcctgcggg gtgcctcggc
     2341 ccctccagcc ccgggccgga cctcctgctg gctctcgccc accaaccaag tgttacaagc
     2401 cccagaatgc tgcccggcct gccctgctgg gcggactgtc tgtgtgtctg tctctctggc
     2461 gttccacctc caagcctata ccagctgtgt acagcgccat ctctctgcct tctgttgccc
     2521 ctcactcacc aaacacgtgt atttatagca aaagtcgacc tgcaggcatg caagctgatc
     2581 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     2641 tagcataacc ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt
     2701 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2761 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2821 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     2881 tacaaatgtg gtaaaatcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     2941 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac
     3001 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga
     3061 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3241 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3301 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3361 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg
     3421 acacaacagt ctcgaaccaa aggctggagc caccatggct tccaaggtgt acgaccccga
     3481 gcaacgcaaa cgcatgatca ctgggcctca gtggtgggct cgctgcaagc aaatgaacgt
     3541 gctggactcc ttcatcaact actatgattc cgagaagcac gccgagaacg ccgtgatttt
     3601 tctgcatggt aacgctgcct ccagctacct gtggaggcac gtcgtgcctc acatcgagcc
     3661 cgtggctaga tgcatcatcc ctgatctgat cggaatgggt aagtccggca agagcgggaa
     3721 tggctcatat cgcctcctgg atcactacaa gtacctcacc gcttggttcg agctgctgaa
     3781 ccttccaaag aaaatcatct ttgtgggcca cgactggggg gcttgtctgg cctttcacta
     3841 ctcctacgag caccaagaca agatcaaggc catcgtccat gctgagagtg tcgtggacgt
     3901 gatcgagtcc tgggacgagt ggcctgacat cgaggaggat atcgccctga tcaagagcga
     3961 agagggcgag aaaatggtgc ttgagaataa cttcttcgtc gagaccatgc tcccaagcaa
     4021 gatcatgcgg aaactggagc ctgaggagtt cgctgcctac ctggagccat tcaaggagaa
     4081 gggcgaggtt agacggccta ccctctcctg gcctcgcgag atccctctcg ttaagggagg
     4141 caagcccgac gtcgtccaga ttgtccgcaa ctacaacgcc taccttcggg ccagcgacga
     4201 tctgcctaag atgttcatcg agtccgaccc tgggttcttt tccaacgcta ttgtcgaggg
     4261 agctaagaag ttccctaaca ccgagttcgt gaaggtgaag ggcctccact tcagccagga
     4321 ggacgctcca gatgaaatgg gtaagtacat caagagcttc gtggagcgcg tgctgaagaa
     4381 cgagcagacc ggtggtggga gcggaggtgg cggatcaggt ggcggaggct ccggagggat
     4441 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4501 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4561 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4621 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4681 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4741 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4801 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4861 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4921 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4981 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5041 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5101 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5161 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5221 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5281 tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     5341 tgaatcgata gcgataagga tcctctttgc gcttgcgttt tcccttgtcc agatagccca
     5401 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtaatggt
     5461 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5521 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5581 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5641 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5701 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5761 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt tcaaagttct
     5821 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5881 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5941 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6001 caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6061 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6121 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6181 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6241 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6301 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6361 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6421 acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa tgaccgacca
     6481 agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6541 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6601 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6661 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6721 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6781 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6841 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6901 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6961 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7021 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7081 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7141 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7201 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7261 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7321 tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt gaaaccatga
     7381 aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata
     7441 ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt
     7501 gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat
     7561 taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga
//
