LOCUS       sequence-231314-        8305 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 8305)
  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 8305)
  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-CERS2-201
FEATURES             Location/Qualifiers
     source          1..8305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        67..566
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(102..125)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     499..518
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..2265
                     /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"
     CDS             2793..2804
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3388..3509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3399..3418)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3453..3472
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3754..4111
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3754..3945
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3777..3797)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3962..4097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4156..4165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4162..5094
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             5146..5937
                     /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(5197..5216)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5807..5826
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6001..6049
                     /note="synthetic polyadenylation signal"
     promoter        6205..6309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6310..6378,6379..7170)
                     /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(6528..6547)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7328..7916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7408..7427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(8032..8305,1..10)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
       61 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
      121 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
      181 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
      241 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
      301 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
      361 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
      421 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
      481 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
      541 cctcttcctc atctccgggc ctttcgacct gcagcccaag cttggcaatc cggtactgtt
      601 ggtaaagcca ccatggaaga tgccaaaaac attaagaagg gcccagcgcc attctaccca
      661 ctcgaagacg ggaccgccgg cgagcagctg cacaaagcca tgaagcgcta cgccctggtg
      721 cccggcacca tcgcctttac cgacgcacat atcgaggtgg acattaccta cgccgagtac
      781 ttcgagatga gcgttcggct ggcagaagct atgaagcgct atgggctgaa tacaaaccat
      841 cggatcgtgg tgtgcagcga gaatagcttg cagttcttca tgcccgtgtt gggtgccctg
      901 ttcatcggtg tggctgtggc cccagctaac gacatctaca acgagcgcga gctgctgaac
      961 agcatgggca tcagccagcc caccgtcgta ttcgtgagca agaaagggct gcaaaagatc
     1021 ctcaacgtgc aaaagaagct accgatcata caaaagatca tcatcatgga tagcaagacc
     1081 gactaccagg gcttccaaag catgtacacc ttcgtgactt cccatttgcc acccggcttc
     1141 aacgagtacg acttcgtgcc cgagagcttc gaccgggaca aaaccatcgc cctgatcatg
     1201 aacagtagtg gcagtaccgg attgcccaag ggcgtagccc taccgcaccg caccgcttgt
     1261 gtccgattca gtcatgcccg cgaccccatc ttcggcaacc agatcatccc cgacaccgct
     1321 atcctcagcg tggtgccatt tcaccacggc ttcggcatgt tcaccacgct gggctacttg
     1381 atctgcggct ttcgggtcgt gctcatgtac cgcttcgagg aggagctatt cttgcgcagc
     1441 ttgcaagact ataagattca atctgccctg ctggtgccca cactatttag cttcttcgct
     1501 aagagcactc tcatcgacaa gtacgaccta agcaacttgc acgagatcgc cagcggcggg
     1561 gcgccgctca gcaaggaggt aggtgaggcc gtggccaaac gcttccacct accaggcatc
     1621 cgccagggct acggcctgac agaaacaacc agcgccattc tgatcacccc cgaaggggac
     1681 gacaagcctg gcgcagtagg caaggtggtg cccttcttcg aggctaaggt ggtggacttg
     1741 gacaccggta agacactggg tgtgaaccag cgcggcgagc tgtgcgtccg tggccccatg
     1801 atcatgagcg gctacgttaa caaccccgag gctacaaacg ctctcatcga caaggacggc
     1861 tggctgcaca gcggcgacat cgcctactgg gacgaggacg agcacttctt catcgtggac
     1921 cggctgaaga gcctgatcaa atacaagggc taccaggtag ccccagccga actggagagc
     1981 atcctgctgc aacaccccaa catcttcgac gccggggtcg ccggcctgcc cgacgacgat
     2041 gccggcgagc tgcccgccgc agtcgtcgtg ctggaacacg gtaaaaccat gaccgagaag
     2101 gagatcgtgg actatgtggc cagccaggtt acaaccgcca agaagctgcg cggtggtgtt
     2161 gtgttcgtgg acgaggtgcc taaaggactg accggcaagt tggacgcccg caagatccgc
     2221 gagattctca ttaaggccaa gaagggcggc aagatcgccg tgtaattcta gttgtttaaa
     2281 cgagaccatt actccagctg cctcccagat taatgcataa agccaaggaa ctaccccgct
     2341 ccctgcgcta tagggtcact ttaagctctg gggaaaaagg agaaagtgag aggagagttc
     2401 tctgcatcct ccctccttgc ttgtcaccca gttgccttta aaccaaattc taaccagcct
     2461 atccccaggt agggggacgt tggttatatt ctgttagagg gggacggtcg tattttcctc
     2521 cctacccgcc aagtcatcct ttctactgct tttgaggccc tccctcagct ctctgtgggt
     2581 aggggttaca attcacattc cttattctga gaatttggcc ccagctgttt gcctttgact
     2641 ccctgacctc cagagccagg gttgtgcctt attgtcccat ctgtgggcct cattctgcca
     2701 aagctggacc aaggctaacc tttctaagct ccctaacttg ggccagaaac caaagctgag
     2761 cttttaactt tctccctcta tgacacaaat gaattgaggg taggaggagg gtgcacataa
     2821 cccttaccct acctctgcca aaaagtgggg gctgtactgg ggactgctcg gatgatcttt
     2881 cttagtgcta cttctttcag ctgtccctgt agcgacaggt ctaagatctg actgcctcct
     2941 cctttctctg gcctcttccc ccttccctct tctcttcagc taggctagct ggtttggagt
     3001 agaatggcaa ctaattctaa tttttattta ttaaatattt ggggttttgg ttttaaagcc
     3061 agaattacgg ctagcaccta gcatttcagc agagggacca ttttagacca aaatgtactg
     3121 ttaatgggtt tttttttaaa attaaaagat taaataaaaa atattaaata aaacatggca
     3181 ataagtgtca gactattagg aattgagaag ggggatcaac taaataaacg aagagagtct
     3241 ttcttatgcc ttccttgcaa gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa
     3301 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     3361 tggggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3421 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3481 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3541 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     3601 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     3661 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     3721 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     3781 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3841 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3901 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3961 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4021 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4081 aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc
     4141 gaaccaaagg ctggagccac catggcttcc aaggtgtacg accccgagca acgcaaacgc
     4201 atgatcactg ggcctcagtg gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc
     4261 atcaactact atgattccga gaagcacgcc gagaacgccg tgatttttct gcatggtaac
     4321 gctgcctcca gctacctgtg gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc
     4381 atcatccctg atctgatcgg aatgggtaag tccggcaaga gcgggaatgg ctcatatcgc
     4441 ctcctggatc actacaagta cctcaccgct tggttcgagc tgctgaacct tccaaagaaa
     4501 atcatctttg tgggccacga ctggggggct tgtctggcct ttcactactc ctacgagcac
     4561 caagacaaga tcaaggccat cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg
     4621 gacgagtggc ctgacatcga ggaggatatc gccctgatca agagcgaaga gggcgagaaa
     4681 atggtgcttg agaataactt cttcgtcgag accatgctcc caagcaagat catgcggaaa
     4741 ctggagcctg aggagttcgc tgcctacctg gagccattca aggagaaggg cgaggttaga
     4801 cggcctaccc tctcctggcc tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc
     4861 gtccagattg tccgcaacta caacgcctac cttcgggcca gcgacgatct gcctaagatg
     4921 ttcatcgagt ccgaccctgg gttcttttcc aacgctattg tcgagggagc taagaagttc
     4981 cctaacaccg agttcgtgaa ggtgaagggc ctccacttca gccaggagga cgctccagat
     5041 gaaatgggta agtacatcaa gagcttcgtg gagcgcgtgc tgaagaacga gcagaccggt
     5101 ggtgggagcg gaggtggcgg atcaggtggc ggaggctccg gagggattga acaagatgga
     5161 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5221 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5281 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5341 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5401 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5461 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5521 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5581 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5641 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5701 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5761 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5821 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5881 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5941 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     6001 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     6061 ataaggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     6121 atccggggtc agcaccgttt ctgcggactg gctttctacg taatggtttc ttagacgtca
     6181 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6241 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6301 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6361 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6421 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6481 tttcgccccg aagaacgttt tccaatgatg agcactttca aagttctgct atgtggcgcg
     6541 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6601 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6661 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6721 acaactatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6781 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6841 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6901 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6961 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7021 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7081 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7141 ataggtgcct cactgattaa gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac
     7201 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7501 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7741 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     7921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata
     8041 caggaacgca cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt
     8101 cagtggatta agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg
     8161 ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct
     8221 gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg
     8281 taaatccact tactgtctgc cctcg
//
