LOCUS       sequence-231735-        7625 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 7625)
  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 7625)
  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-LFNG-203
FEATURES             Location/Qualifiers
     source          1..7625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..534
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(70..93)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     467..486
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      575..584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             581..2233
                     /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    2676..2797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2687..2706)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2741..2760
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3042..3399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3042..3233
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3065..3085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3250..3385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3444..3453
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3450..4382
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4434..5225
                     /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(4485..4504)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5095..5114
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5289..5337
                     /note="synthetic polyadenylation signal"
     promoter        5493..5597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5598..5666,5667..6458)
                     /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(5816..5835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6616..7204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6696..6715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7320..7603
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 acgatgcgtc cggcgtagag gatcgagatc taccgggtag gggaggcgct tttcccaagg
       61 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
      121 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
      181 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
      241 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
      301 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
      361 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
      421 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
      481 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc
      541 agcccaagct tggcaatccg gtactgttgg taaagccacc atggaagatg ccaaaaacat
      601 taagaagggc ccagcgccat tctacccact cgaagacggg accgccggcg agcagctgca
      661 caaagccatg aagcgctacg ccctggtgcc cggcaccatc gcctttaccg acgcacatat
      721 cgaggtggac attacctacg ccgagtactt cgagatgagc gttcggctgg cagaagctat
      781 gaagcgctat gggctgaata caaaccatcg gatcgtggtg tgcagcgaga atagcttgca
      841 gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc cagctaacga
      901 catctacaac gagcgcgagc tgctgaacag catgggcatc agccagccca ccgtcgtatt
      961 cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa aagaagctac cgatcataca
     1021 aaagatcatc atcatggata gcaagaccga ctaccagggc ttccaaagca tgtacacctt
     1081 cgtgacttcc catttgccac ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga
     1141 ccgggacaaa accatcgccc tgatcatgaa cagtagtggc agtaccggat tgcccaaggg
     1201 cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg accccatctt
     1261 cggcaaccag atcatccccg acaccgctat cctcagcgtg gtgccatttc accacggctt
     1321 cggcatgttc accacgctgg gctacttgat ctgcggcttt cgggtcgtgc tcatgtaccg
     1381 cttcgaggag gagctattct tgcgcagctt gcaagactat aagattcaat ctgccctgct
     1441 ggtgcccaca ctatttagct tcttcgctaa gagcactctc atcgacaagt acgacctaag
     1501 caacttgcac gagatcgcca gcggcggggc gccgctcagc aaggaggtag gtgaggccgt
     1561 ggccaaacgc ttccacctac caggcatccg ccagggctac ggcctgacag aaacaaccag
     1621 cgccattctg atcacccccg aaggggacga caagcctggc gcagtaggca aggtggtgcc
     1681 cttcttcgag gctaaggtgg tggacttgga caccggtaag acactgggtg tgaaccagcg
     1741 cggcgagctg tgcgtccgtg gccccatgat catgagcggc tacgttaaca accccgaggc
     1801 tacaaacgct ctcatcgaca aggacggctg gctgcacagc ggcgacatcg cctactggga
     1861 cgaggacgag cacttcttca tcgtggaccg gctgaagagc ctgatcaaat acaagggcta
     1921 ccaggtagcc ccagccgaac tggagagcat cctgctgcaa caccccaaca tcttcgacgc
     1981 cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg cccgccgcag tcgtcgtgct
     2041 ggaacacggt aaaaccatga ccgagaagga gatcgtggac tatgtggcca gccaggttac
     2101 aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta aaggactgac
     2161 cggcaagttg gacgcccgca agatccgcga gattctcatt aaggccaaga agggcggcaa
     2221 gatcgccgtg taattctagt tgtttaaacg agctcgggca cggagcagct gatgcccaag
     2281 gtccagcagc catgctggtg ggctggggag gcgctcagac tccaagccag gcaggccgtg
     2341 ggtccggccg ccccacagct ggtaatgctg cacctccctc tcctgtgatg tggccccagg
     2401 ccccgacccc gctcaggggt ggtgggaaga gtcctggaga gcaggagcag tgccggagcc
     2461 cccactgcag ctggtcccag cccctgcaga gatccagacg cttgtttttt aaatacagca
     2521 gctctgcggt gcttttcctt cctcaaaagt cgacctgcag gcatgcaagc tgatccggct
     2581 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     2641 taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2701 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2761 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2821 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     2881 atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     2941 tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg
     3001 cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag
     3061 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3301 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgattc ttctgacaca
     3421 acagtctcga accaaaggct ggagccacca tggcttccaa ggtgtacgac cccgagcaac
     3481 gcaaacgcat gatcactggg cctcagtggt gggctcgctg caagcaaatg aacgtgctgg
     3541 actccttcat caactactat gattccgaga agcacgccga gaacgccgtg atttttctgc
     3601 atggtaacgc tgcctccagc tacctgtgga ggcacgtcgt gcctcacatc gagcccgtgg
     3661 ctagatgcat catccctgat ctgatcggaa tgggtaagtc cggcaagagc gggaatggct
     3721 catatcgcct cctggatcac tacaagtacc tcaccgcttg gttcgagctg ctgaaccttc
     3781 caaagaaaat catctttgtg ggccacgact ggggggcttg tctggccttt cactactcct
     3841 acgagcacca agacaagatc aaggccatcg tccatgctga gagtgtcgtg gacgtgatcg
     3901 agtcctggga cgagtggcct gacatcgagg aggatatcgc cctgatcaag agcgaagagg
     3961 gcgagaaaat ggtgcttgag aataacttct tcgtcgagac catgctccca agcaagatca
     4021 tgcggaaact ggagcctgag gagttcgctg cctacctgga gccattcaag gagaagggcg
     4081 aggttagacg gcctaccctc tcctggcctc gcgagatccc tctcgttaag ggaggcaagc
     4141 ccgacgtcgt ccagattgtc cgcaactaca acgcctacct tcgggccagc gacgatctgc
     4201 ctaagatgtt catcgagtcc gaccctgggt tcttttccaa cgctattgtc gagggagcta
     4261 agaagttccc taacaccgag ttcgtgaagg tgaagggcct ccacttcagc caggaggacg
     4321 ctccagatga aatgggtaag tacatcaaga gcttcgtgga gcgcgtgctg aagaacgagc
     4381 agaccggtgg tgggagcgga ggtggcggat caggtggcgg aggctccgga gggattgaac
     4441 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4501 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4561 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4621 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4681 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4741 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4801 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4861 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4921 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4981 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5041 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5101 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5161 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5221 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5281 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     5341 cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     5401 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgta atggtttctt
     5461 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5521 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5581 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5641 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5701 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5761 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cactttcaaa gttctgctat
     5821 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5881 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5941 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6001 tacttctgac aactatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6061 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6121 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6181 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6241 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6301 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6361 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6421 tcgctgagat aggtgcctca ctgattaagc attggtaatt cgaaatgacc gaccaagcga
     6481 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6541 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6601 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6661 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6721 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6781 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6841 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6901 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6961 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7021 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7081 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7141 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     7201 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7261 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     7321 cggaaataca ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat
     7381 ggcagcttca gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat
     7441 tcatacggtt aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat
     7501 ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg
     7561 gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc
     7621 cggcc
//
