LOCUS       sequence-231684-        8429 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 8429)
  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 8429)
  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-FKBP1A-204
FEATURES             Location/Qualifiers
     source          1..8429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..562
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(98..121)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     495..514
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      603..612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             609..2261
                     /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    3508..3629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3519..3538)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3573..3592
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3874..4231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3874..4065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3897..3917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4082..4217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4276..4285
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4282..5214
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             5266..6057
                     /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(5317..5336)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5927..5946
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6121..6169
                     /note="synthetic polyadenylation signal"
     promoter        6325..6429
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6430..6498,6499..7290)
                     /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(6648..6667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7448..8036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7528..7547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(8152..8429,1..6)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatcta
       61 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
      121 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
      181 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
      241 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
      301 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
      361 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
      421 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
      481 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
      541 ttcctcatct ccgggccttt cgacctgcag cccaagcttg gcaatccggt actgttggta
      601 aagccaccat ggaagatgcc aaaaacatta agaagggccc agcgccattc tacccactcg
      661 aagacgggac cgccggcgag cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg
      721 gcaccatcgc ctttaccgac gcacatatcg aggtggacat tacctacgcc gagtacttcg
      781 agatgagcgt tcggctggca gaagctatga agcgctatgg gctgaataca aaccatcgga
      841 tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt gccctgttca
      901 tcggtgtggc tgtggcccca gctaacgaca tctacaacga gcgcgagctg ctgaacagca
      961 tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa aagatcctca
     1021 acgtgcaaaa gaagctaccg atcatacaaa agatcatcat catggatagc aagaccgact
     1081 accagggctt ccaaagcatg tacaccttcg tgacttccca tttgccaccc ggcttcaacg
     1141 agtacgactt cgtgcccgag agcttcgacc gggacaaaac catcgccctg atcatgaaca
     1201 gtagtggcag taccggattg cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc
     1261 gattcagtca tgcccgcgac cccatcttcg gcaaccagat catccccgac accgctatcc
     1321 tcagcgtggt gccatttcac cacggcttcg gcatgttcac cacgctgggc tacttgatct
     1381 gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga gctattcttg cgcagcttgc
     1441 aagactataa gattcaatct gccctgctgg tgcccacact atttagcttc ttcgctaaga
     1501 gcactctcat cgacaagtac gacctaagca acttgcacga gatcgccagc ggcggggcgc
     1561 cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc
     1621 agggctacgg cctgacagaa acaaccagcg ccattctgat cacccccgaa ggggacgaca
     1681 agcctggcgc agtaggcaag gtggtgccct tcttcgaggc taaggtggtg gacttggaca
     1741 ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca
     1801 tgagcggcta cgttaacaac cccgaggcta caaacgctct catcgacaag gacggctggc
     1861 tgcacagcgg cgacatcgcc tactgggacg aggacgagca cttcttcatc gtggaccggc
     1921 tgaagagcct gatcaaatac aagggctacc aggtagcccc agccgaactg gagagcatcc
     1981 tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg
     2041 gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga
     2101 tcgtggacta tgtggccagc caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt
     2161 tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag atccgcgaga
     2221 ttctcattaa ggccaagaag ggcggcaaga tcgccgtgta attctagttg tttaaacgag
     2281 ctcgatctgc catggaggga tctggtgcct ccagacatgt gcacatgaat ccatatggag
     2341 cttttcctga tgttccactc cactttgtat agacatctgc cctgactgaa tgtgttctgt
     2401 cactcagctt tgcttccgac acctctgttt cctcttcccc tttctcctcg tatgtgtgtt
     2461 tacctaaact atatgccata aacctcaagt tattcatttt attttgtttt cattttgggg
     2521 tgaagattca gtttcagtct tttggatata ggtttccaat taagtacatg gtcaagtatt
     2581 aacagcacaa gtggtaggtt aacattagaa taggaattgg tgttgggggg ggggtttgca
     2641 agaatatttt attttaattt tttggatgaa atttttatct attatatatt aaacattctt
     2701 gctgctgcgc tgcaaagcca tagcagattt gaggcgctgt tgaggactga attactctcc
     2761 aagttgagag atgtctttgg gttaaattaa aagccctacc taaaactgag gtggggatgg
     2821 ggagagcctt tgcctccacc attcccaccc accctcccct taaaccctct gcctttgaaa
     2881 gtagatcatg ttcactgcaa tgctggacac tacaggtatc tgtccctggg ccagcaggga
     2941 cctctgaagc cttctttgtg gccttttttt tttttcatcc tgtggttttt ctaatggact
     3001 ttcaggaatt ttgtaatctc ataactttcc aagctccacc acttcctaaa tcttaagaac
     3061 tttaattgac agtttcaatt gaaggtgctg tttgtagact taacacccag tgaaagccca
     3121 gccatcatga caaatccttg aatgttctct taagaaaatg atgctggtca tcgcagcttc
     3181 agcatctcct gttttttgat gcttggctcc ctctgctgat ctcagtttcc tggcttttcc
     3241 tccctcagcc ccttctcacc cctttgctgt cctgtgtagt gatttggtga gaaatcgttg
     3301 ctgcaccctt cccccagcac catttatgag tctcaagttt tattattgca ataaaagtgc
     3361 tttatgccgg cttttctcaa gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     3421 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     3481 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3541 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3601 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3661 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     3721 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     3781 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     3841 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc
     4261 gaaccaaagg ctggagccac catggcttcc aaggtgtacg accccgagca acgcaaacgc
     4321 atgatcactg ggcctcagtg gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc
     4381 atcaactact atgattccga gaagcacgcc gagaacgccg tgatttttct gcatggtaac
     4441 gctgcctcca gctacctgtg gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc
     4501 atcatccctg atctgatcgg aatgggtaag tccggcaaga gcgggaatgg ctcatatcgc
     4561 ctcctggatc actacaagta cctcaccgct tggttcgagc tgctgaacct tccaaagaaa
     4621 atcatctttg tgggccacga ctggggggct tgtctggcct ttcactactc ctacgagcac
     4681 caagacaaga tcaaggccat cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg
     4741 gacgagtggc ctgacatcga ggaggatatc gccctgatca agagcgaaga gggcgagaaa
     4801 atggtgcttg agaataactt cttcgtcgag accatgctcc caagcaagat catgcggaaa
     4861 ctggagcctg aggagttcgc tgcctacctg gagccattca aggagaaggg cgaggttaga
     4921 cggcctaccc tctcctggcc tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc
     4981 gtccagattg tccgcaacta caacgcctac cttcgggcca gcgacgatct gcctaagatg
     5041 ttcatcgagt ccgaccctgg gttcttttcc aacgctattg tcgagggagc taagaagttc
     5101 cctaacaccg agttcgtgaa ggtgaagggc ctccacttca gccaggagga cgctccagat
     5161 gaaatgggta agtacatcaa gagcttcgtg gagcgcgtgc tgaagaacga gcagaccggt
     5221 ggtgggagcg gaggtggcgg atcaggtggc ggaggctccg gagggattga acaagatgga
     5281 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5341 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5401 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5461 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5521 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5581 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5641 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5701 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5761 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5821 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5881 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5941 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6001 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6061 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     6121 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     6181 ataaggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     6241 atccggggtc agcaccgttt ctgcggactg gctttctacg taatggtttc ttagacgtca
     6301 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6361 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6421 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6481 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6541 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6601 tttcgccccg aagaacgttt tccaatgatg agcactttca aagttctgct atgtggcgcg
     6661 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6721 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6781 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6841 acaactatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6901 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6961 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7021 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7081 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7141 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7201 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7261 ataggtgcct cactgattaa gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac
     7321 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7381 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7861 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     8041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata
     8161 caggaacgca cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt
     8221 cagtggatta agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg
     8281 ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct
     8341 gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg
     8401 taaatccact tactgtctgc cctcgtagc
//
