LOCUS       sequence-229335-        8846 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 8846)
  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 8846)
  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-CTDSP1-201
FEATURES             Location/Qualifiers
     source          1..8846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    110..393
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        450..949
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(485..508)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     882..901
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      990..999
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             996..2648
                     /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    4312..4433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4323..4342)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4377..4396
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4678..5035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4678..4869
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4701..4721)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4886..5021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      5080..5089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5086..6018
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             6070..6861
                     /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(6121..6140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6731..6750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6925..6973
                     /note="synthetic polyadenylation signal"
     promoter        7129..7233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7234..7302,7303..8094)
                     /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(7452..7471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(8332..8351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
       61 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca
      121 ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca
      181 gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt
      241 aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt
      301 ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta
      361 aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat
      421 gcgtccggcg tagaggatcg agatctaccg ggtaggggag gcgcttttcc caaggcagtc
      481 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
      541 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
      601 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
      661 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
      721 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
      781 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
      841 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
      901 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc
      961 aagcttggca atccggtact gttggtaaag ccaccatgga agatgccaaa aacattaaga
     1021 agggcccagc gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag
     1081 ccatgaagcg ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg
     1141 tggacattac ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc
     1201 gctatgggct gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct
     1261 tcatgcccgt gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct
     1321 acaacgagcg cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga
     1381 gcaagaaagg gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga
     1441 tcatcatcat ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga
     1501 cttcccattt gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg
     1561 acaaaaccat cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag
     1621 ccctaccgca ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca
     1681 accagatcat ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca
     1741 tgttcaccac gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg
     1801 aggaggagct attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc
     1861 ccacactatt tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact
     1921 tgcacgagat cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca
     1981 aacgcttcca cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca
     2041 ttctgatcac ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct
     2101 tcgaggctaa ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg
     2161 agctgtgcgt ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa
     2221 acgctctcat cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg
     2281 acgagcactt cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg
     2341 tagccccagc cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg
     2401 tcgccggcct gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac
     2461 acggtaaaac catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg
     2521 ccaagaagct gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca
     2581 agttggacgc ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg
     2641 ccgtgtaatt ctagttgttt aaacgagtga gggtgatggg gccaggacct gcccctgacc
     2701 aatgataccc acacctcctc ccaggaagac tgcccaggcc tttgttagga aaacccatgg
     2761 gccgccgcca cactcagtgc catggggaag cgggcgtctc ccccaccagc cccaccaggc
     2821 ggtgtagggg cagcaggctg cactgaggac cgtgagctcc aggccccgtg tcagtgcctt
     2881 caaacctcct cccctattct caggggacct ggggggccct gcctgctgct ccctttttct
     2941 gtctctgtcc atgctgccat gtttctctgc tgccaaattg ggccccttgg ccccttccgg
     3001 ttctgcttcc tgggggcagg gttcctgcct tggaccccca gtctgggaac ggtggacatc
     3061 aagtaccttg catagagccc cctcttcccc gcccagcttt cccaggggca cagctctagg
     3121 ctgggagggg agaaccagcc cctccccctg ccccacctcc tcccttggga ctgagagggc
     3181 ccctaccaac ctttgcctct gccttggagg gaggggaggt ctgttaccac tggggaaggc
     3241 agcaggagtc tgtccttcag gccccacagt gcagcttctc cagggccgac agctgagggc
     3301 tgctccctgc atcatccaag caatgacctc agacttctgc cttaaccagc cccggggctt
     3361 ggctccccca gctctgagcg tgggggcata ggcaggaccc cccttgtggt gccatataaa
     3421 tatgtacatg tgtatataga tttttagggg aaggagagag ggaagggtca gggtagagac
     3481 acccctccct tgcccctttc ctgggcccag aagttggggg gagggaggga aaggattttt
     3541 acatttttta aactgctatt ttctgaatgg aacaagctgg gccaaggggc ccaggccctg
     3601 tcctctgtcc ctcacacccc tttgctccgt tcattcattc aaaaaaacat ttcttgagca
     3661 ccttctgtgc ccagcatatg ctaggcccac cagctaagtg tgtgtggggg gtctctacgc
     3721 cagctcatca gtgcctcctt gcccatcctt caccggtgcc tttgggggat ctgtaggagg
     3781 tgggaccttc tgtggggttt ggggatctcc aggaagcccg accaagctgt ccccttcccc
     3841 tgtgccaacc catctcctac agccccctgc ctgatcccct gctggctggg ggcagctccc
     3901 aggatatcct gccttccaac tgtttctgaa gcccctcctc ctaacatggc gattccggag
     3961 gtcaaggcct tgggctctcc ccagggtcta acggttaagg ggacccacat accagtgcca
     4021 agggggatgt caagtggtga tgtcgttgtg ctcccctccc ccagagcggg tgggcggggg
     4081 gtgaatatgg ttggcctgca tcaggtggcc ttcccattta agtgccttct ctgtgactga
     4141 gagccctagt gtgatgagaa ctaaagagaa agccagaccc ctaagtcgac ctgcaggcat
     4201 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     4261 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     4321 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     4381 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     4441 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     4501 agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt
     4561 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     4621 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     4681 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4741 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4861 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4921 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4981 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5041 tgattcttct gacacaacag tctcgaacca aaggctggag ccaccatggc ttccaaggtg
     5101 tacgaccccg agcaacgcaa acgcatgatc actgggcctc agtggtgggc tcgctgcaag
     5161 caaatgaacg tgctggactc cttcatcaac tactatgatt ccgagaagca cgccgagaac
     5221 gccgtgattt ttctgcatgg taacgctgcc tccagctacc tgtggaggca cgtcgtgcct
     5281 cacatcgagc ccgtggctag atgcatcatc cctgatctga tcggaatggg taagtccggc
     5341 aagagcggga atggctcata tcgcctcctg gatcactaca agtacctcac cgcttggttc
     5401 gagctgctga accttccaaa gaaaatcatc tttgtgggcc acgactgggg ggcttgtctg
     5461 gcctttcact actcctacga gcaccaagac aagatcaagg ccatcgtcca tgctgagagt
     5521 gtcgtggacg tgatcgagtc ctgggacgag tggcctgaca tcgaggagga tatcgccctg
     5581 atcaagagcg aagagggcga gaaaatggtg cttgagaata acttcttcgt cgagaccatg
     5641 ctcccaagca agatcatgcg gaaactggag cctgaggagt tcgctgccta cctggagcca
     5701 ttcaaggaga agggcgaggt tagacggcct accctctcct ggcctcgcga gatccctctc
     5761 gttaagggag gcaagcccga cgtcgtccag attgtccgca actacaacgc ctaccttcgg
     5821 gccagcgacg atctgcctaa gatgttcatc gagtccgacc ctgggttctt ttccaacgct
     5881 attgtcgagg gagctaagaa gttccctaac accgagttcg tgaaggtgaa gggcctccac
     5941 ttcagccagg aggacgctcc agatgaaatg ggtaagtaca tcaagagctt cgtggagcgc
     6001 gtgctgaaga acgagcagac cggtggtggg agcggaggtg gcggatcagg tggcggaggc
     6061 tccggaggga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6121 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6181 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6241 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6301 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6361 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6421 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6481 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6541 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6601 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6661 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6721 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6781 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6841 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6901 ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     6961 ggttttttgt gtgaatcgat agcgataagg atcctctttg cgcttgcgtt ttcccttgtc
     7021 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     7081 tacgtaatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7141 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7201 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7261 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7321 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7381 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7441 ttcaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     7501 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     7561 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7621 aacactgcgg ccaacttact tctgacaact atcggaggac cgaaggagct aaccgctttt
     7681 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7741 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     7801 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7861 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7921 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7981 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8041 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaattcgaa
     8101 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     8161 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     8221 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     8281 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     8341 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     8401 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     8461 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     8521 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     8581 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     8641 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     8701 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     8761 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     8821 gcagaaaaaa aggatttcaa gaagat
//
