LOCUS       sequence-231734-        8551 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 8551)
  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 8551)
  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-201_UTR
FEATURES             Location/Qualifiers
     source          1..8551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        38..537
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(73..96)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     470..489
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      578..587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             584..2236
                     /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    3605..3726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3616..3635)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3670..3689
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3971..4328
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3971..4162
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3994..4014)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4179..4314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4373..4382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4379..5311
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             5363..6154
                     /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(5414..5433)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6024..6043
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6218..6266
                     /note="synthetic polyadenylation signal"
     promoter        6422..6526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6527..6595,6596..7387)
                     /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(6745..6764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7545..8133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7625..7644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8249..8532
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 gccacgatgc gtccggcgta gaggatcgag atctaccggg taggggaggc gcttttccca
       61 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
      121 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
      181 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
      241 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
      301 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
      361 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
      421 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
      481 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc
      541 tgcagcccaa gcttggcaat ccggtactgt tggtaaagcc accatggaag atgccaaaaa
      601 cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg gcgagcagct
      661 gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta ccgacgcaca
      721 tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc tggcagaagc
      781 tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg agaatagctt
      841 gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg ccccagctaa
      901 cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc ccaccgtcgt
      961 attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc taccgatcat
     1021 acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa gcatgtacac
     1081 cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc ccgagagctt
     1141 cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg gattgcccaa
     1201 gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc gcgaccccat
     1261 cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg
     1321 cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg tgctcatgta
     1381 ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc aatctgccct
     1441 gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca agtacgacct
     1501 aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc
     1561 cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac
     1621 cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt
     1681 gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca
     1741 gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga
     1801 ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg
     1861 ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg
     1921 ctaccaggta gccccagccg aactggagag catcctgctg caacacccca acatcttcga
     1981 cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt
     2041 gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     2101 tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact
     2161 gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg
     2221 caagatcgcc gtgtaattct agttgtttaa acgagctcgt ggccatggct gagacccaat
     2281 ccctgggcgc ccctggtatc caaagggccc agggaccctg ttgcgctgcc ctggcctcgg
     2341 cattcgaggc tcccctaggg ccgtgcctgt gcgtgtgcgt gtgcgtgtgt gtgtgtgtgt
     2401 actgcatgcc cacccgggta gcaggctgct gggcagttct gctctgtgga ggggcgggca
     2461 ccagcgccac ttatgtgcct ctgctccgag ggccagtggg ctgcagggcc tgcttggagg
     2521 aaggatttgt gtgtcggagg ccactccgag ggcaattctg ttaggatttt tggatctttc
     2581 tacagctacg gggctccggg ctactttgca gggatgcgat gcgtaggtgc ctttctcttc
     2641 ctgctgacca caagctctgt gctgggggta cctgtgccct gaagtcctgg cccctgtgtc
     2701 atagccccaa gtacgactca ctgagccatg ctcattgcag gggaggctgg gtctgggggt
     2761 gcgtgaccca atccccttcc tcctggcctg gacgctgtgg cttaagagta acagcagcca
     2821 ccgcccagtt ccagtggccc cacgaagccc ccagtggctg gctgtccagc tgggcaaaca
     2881 gtggcacccc tcccagctct tctgagtggg gagtcttcca ggcctcttca gaggtcttcc
     2941 ctttgcctcc ccaggacagg gtgagtcaga gctcagcatt taatctcctc tccaaagtag
     3001 agagcaagtc gcccacagtg ggcgtgtctg taaatattgt gacagtattt ttttactgtg
     3061 ctgttttttt tgaaagggga tgggtaaaaa gtagggtgtt cttgtttttt gcttgggagg
     3121 gtgggggtgg gggagggtct tattttatct tatcttttct gtggatcaga aaaaacagaa
     3181 gccaaactcg gggtcatctt tgtttttaaa gctgaagtgg gactgtctgg cactctgtgt
     3241 atttatgcgt tccagcatct ggaacctccc atccctgccc tcctcctgtg tagctgccac
     3301 ctccccgctg ggcccagcat ggctcacctg tcccgtgggc tgtgtttctt gttgtttttc
     3361 tctttgcaaa gacatagcta ggaaagcgaa tgataaggga aaagttctca gggaattgaa
     3421 gtgttgttgc tatggtgacg tccttttgct gtgaataaag gtgctctttg cagcaaagtc
     3481 gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     3541 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     3601 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3661 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3721 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     3781 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt
     3841 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3901 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     3961 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4021 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4081 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4141 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4201 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4261 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4321 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa ccaaaggctg gagccaccat
     4381 ggcttccaag gtgtacgacc ccgagcaacg caaacgcatg atcactgggc ctcagtggtg
     4441 ggctcgctgc aagcaaatga acgtgctgga ctccttcatc aactactatg attccgagaa
     4501 gcacgccgag aacgccgtga tttttctgca tggtaacgct gcctccagct acctgtggag
     4561 gcacgtcgtg cctcacatcg agcccgtggc tagatgcatc atccctgatc tgatcggaat
     4621 gggtaagtcc ggcaagagcg ggaatggctc atatcgcctc ctggatcact acaagtacct
     4681 caccgcttgg ttcgagctgc tgaaccttcc aaagaaaatc atctttgtgg gccacgactg
     4741 gggggcttgt ctggcctttc actactccta cgagcaccaa gacaagatca aggccatcgt
     4801 ccatgctgag agtgtcgtgg acgtgatcga gtcctgggac gagtggcctg acatcgagga
     4861 ggatatcgcc ctgatcaaga gcgaagaggg cgagaaaatg gtgcttgaga ataacttctt
     4921 cgtcgagacc atgctcccaa gcaagatcat gcggaaactg gagcctgagg agttcgctgc
     4981 ctacctggag ccattcaagg agaagggcga ggttagacgg cctaccctct cctggcctcg
     5041 cgagatccct ctcgttaagg gaggcaagcc cgacgtcgtc cagattgtcc gcaactacaa
     5101 cgcctacctt cgggccagcg acgatctgcc taagatgttc atcgagtccg accctgggtt
     5161 cttttccaac gctattgtcg agggagctaa gaagttccct aacaccgagt tcgtgaaggt
     5221 gaagggcctc cacttcagcc aggaggacgc tccagatgaa atgggtaagt acatcaagag
     5281 cttcgtggag cgcgtgctga agaacgagca gaccggtggt gggagcggag gtggcggatc
     5341 aggtggcgga ggctccggag ggattgaaca agatggattg cacgcaggtt ctccggccgc
     5401 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5461 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5521 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5581 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5641 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5701 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5761 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5821 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5881 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5941 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6001 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6061 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6121 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     6181 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     6241 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc
     6301 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     6361 cggactggct ttctacgtaa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6421 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6481 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6541 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6601 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6661 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6721 aatgatgagc actttcaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6781 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6841 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6901 aaccatgagt gataacactg cggccaactt acttctgaca actatcggag gaccgaagga
     6961 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7021 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7081 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7141 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7201 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7261 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7321 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7381 ttggtaattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     7441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7741 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7981 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     8041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     8101 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     8161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     8221 aggatcttca cctagatcct tttatagtcc ggaaatacag gaacgcacgc tggatggccc
     8281 ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag tggattaagt gggggtaatg
     8341 tggcctgtac cctctggttg cataggtatt catacggtta aaatttatca ggcgcgattg
     8401 cggcagtttt tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg atcgcgctga
     8461 acgcgtttta gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct
     8521 cgtagccatc gagataaacc gcagtactcc g
//
