LOCUS       sequence-231316-        8031 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 8031)
  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 8031)
  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-BAX-201
FEATURES             Location/Qualifiers
     source          1..8031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..561
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(97..120)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     494..513
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      602..611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             608..2260
                     /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    3109..3230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3120..3139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3174..3193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3475..3832
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3475..3666
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3498..3518)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3683..3818
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3877..3886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3883..4815
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4867..5658
                     /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(4918..4937)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5528..5547
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5722..5770
                     /note="synthetic polyadenylation signal"
     promoter        5926..6030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6031..6099,6100..6891)
                     /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(6249..6268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7049..7637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7129..7148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(7753..8031,1..5)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatctac
       61 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg
      121 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg
      181 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc
      241 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
      301 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
      361 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
      421 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
      481 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
      541 tcctcatctc cgggcctttc gacctgcagc ccaagcttgg caatccggta ctgttggtaa
      601 agccaccatg gaagatgcca aaaacattaa gaagggccca gcgccattct acccactcga
      661 agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc tggtgcccgg
      721 caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga
      781 gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat
      841 cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat
      901 cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat
      961 gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa
     1021 cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta
     1081 ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga
     1141 gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag
     1201 tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg
     1261 attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct
     1321 cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg
     1381 cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc gcagcttgca
     1441 agactataag attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag
     1501 cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc
     1561 gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca
     1621 gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa
     1681 gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac
     1741 cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat
     1801 gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct
     1861 gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct
     1921 gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct
     1981 gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg
     2041 cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accatgaccg agaaggagat
     2101 cgtggactat gtggccagcc aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt
     2161 cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat
     2221 tctcattaag gccaagaagg gcggcaagat cgccgtgtaa ttctagttgt ttaaacgagt
     2281 caatccccga ttcatctacc ctgctgacct cccagtgacc cctgacctca ctgtgacctt
     2341 gacttgatta gtgccttctg ccctccctgg agcctccact gcctctggaa ttgctcaagt
     2401 tcattgatga ccctctgacc ctagctcttt cctttttttt tttttcccca ctgagaaggg
     2461 gtctcgctat gttgcccagg ttggtctcga actcctggcc tcaagcgatc ctcccgcctc
     2521 agcctctcaa agtgctggga ttacaggtgt gagccaccat gcctggcctg agtccagctc
     2581 tttaatgccc gttcatctca gtcccctgcc cgcaatcctg ccttctggcc tcctccgtcc
     2641 ctgatcccgc ctctgcctgc ccaggggctg cccctggccg agtcactgaa gcgactgatg
     2701 tccctgtctc caggacggcc tcctctccta ctttgggacg cccacgtggc agaccgtgac
     2761 catctttgtg gcgggagtgc tcaccgcctc actcaccatc tggaagaaga tgggctgagg
     2821 cccccagctg ccttggactg tgtttttcct ccataaatta tggcattttt ctgggagggg
     2881 tggggattgg gggacatggg catttttctt acttttgtaa ttattggggg gtgtggggaa
     2941 gagtggtctt gagggggtaa taaacctcct tcgggacaca agtcgacctg caggcatgca
     3001 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     3061 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     3121 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3181 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3241 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     3301 aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc
     3361 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     3421 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     3481 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3541 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3601 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3661 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3721 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3781 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga
     3841 ttcttctgac acaacagtct cgaaccaaag gctggagcca ccatggcttc caaggtgtac
     3901 gaccccgagc aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa
     3961 atgaacgtgc tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc
     4021 gtgatttttc tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac
     4081 atcgagcccg tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag
     4141 agcgggaatg gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag
     4201 ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc
     4261 tttcactact cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc
     4321 gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc
     4381 aagagcgaag agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc
     4441 ccaagcaaga tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc
     4501 aaggagaagg gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt
     4561 aagggaggca agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc
     4621 agcgacgatc tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt
     4681 gtcgagggag ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc
     4741 agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg
     4801 ctgaagaacg agcagaccgg tggtgggagc ggaggtggcg gatcaggtgg cggaggctcc
     4861 ggagggattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4921 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4981 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5041 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5101 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5161 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5221 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5281 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5341 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5401 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5461 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5521 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5581 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5641 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5701 acctgccatc acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt
     5761 tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc ccttgtccag
     5821 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     5881 gtaatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5941 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6001 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6061 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6121 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6181 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttc
     6241 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6301 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6361 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6421 actgcggcca acttacttct gacaactatc ggaggaccga aggagctaac cgcttttttg
     6481 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6541 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6601 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6661 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6721 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6781 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6841 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta attcgaaatg
     6901 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     6961 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7021 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7081 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7141 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7201 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7261 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7321 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7381 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7441 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7501 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7561 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7621 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7681 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7741 tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga
     7801 aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg
     7861 gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt
     7921 ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg
     7981 cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc c
//
