LOCUS       sequence-231455-        8283 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 8283)
  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 8283)
  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-SEMA5B-202
FEATURES             Location/Qualifiers
     source          1..8283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     393..412
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      501..510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             507..2159
                     /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    3260..3381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3271..3290)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3325..3344
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3626..3983
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3626..3817
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3649..3669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3834..3969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4028..4037
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4034..4966
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             5018..5809
                     /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(5069..5088)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5679..5698
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5873..5921
                     /note="synthetic polyadenylation signal"
     promoter        6077..6181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6182..6250,6251..7042)
                     /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(6400..6419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7200..7788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7280..7299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7904..8187
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(8244..8283,1..460)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(join(8279..8283,1..19))
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca
       61 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
      121 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
      181 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca
      241 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg
      301 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg
      361 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc
      421 tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttggc
      481 aatccggtac tgttggtaaa gccaccatgg aagatgccaa aaacattaag aagggcccag
      541 cgccattcta cccactcgaa gacgggaccg ccggcgagca gctgcacaaa gccatgaagc
      601 gctacgccct ggtgcccggc accatcgcct ttaccgacgc acatatcgag gtggacatta
      661 cctacgccga gtacttcgag atgagcgttc ggctggcaga agctatgaag cgctatgggc
      721 tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg
      781 tgttgggtgc cctgttcatc ggtgtggctg tggccccagc taacgacatc tacaacgagc
      841 gcgagctgct gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag
      901 ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat catacaaaag atcatcatca
      961 tggatagcaa gaccgactac cagggcttcc aaagcatgta caccttcgtg acttcccatt
     1021 tgccacccgg cttcaacgag tacgacttcg tgcccgagag cttcgaccgg gacaaaacca
     1081 tcgccctgat catgaacagt agtggcagta ccggattgcc caagggcgta gccctaccgc
     1141 accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc aaccagatca
     1201 tccccgacac cgctatcctc agcgtggtgc catttcacca cggcttcggc atgttcacca
     1261 cgctgggcta cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc
     1321 tattcttgcg cagcttgcaa gactataaga ttcaatctgc cctgctggtg cccacactat
     1381 ttagcttctt cgctaagagc actctcatcg acaagtacga cctaagcaac ttgcacgaga
     1441 tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc
     1501 acctaccagg catccgccag ggctacggcc tgacagaaac aaccagcgcc attctgatca
     1561 cccccgaagg ggacgacaag cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta
     1621 aggtggtgga cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg
     1681 tccgtggccc catgatcatg agcggctacg ttaacaaccc cgaggctaca aacgctctca
     1741 tcgacaagga cggctggctg cacagcggcg acatcgccta ctgggacgag gacgagcact
     1801 tcttcatcgt ggaccggctg aagagcctga tcaaatacaa gggctaccag gtagccccag
     1861 ccgaactgga gagcatcctg ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc
     1921 tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa
     1981 ccatgaccga gaaggagatc gtggactatg tggccagcca ggttacaacc gccaagaagc
     2041 tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg actgaccggc aagttggacg
     2101 cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc gccgtgtaat
     2161 tctagttgtt taaacgagct cgtaccgccg tcctggggac ttgggcttct tgccttcata
     2221 aggcacagag cagatggaga tgggacagtg gagccagttt ggttttctcc ctctgcacta
     2281 ggccaagaac ttgctgcctt gcctgtgggg ggtcccatcc ggcttcagag agctctggct
     2341 ggcattgacc atgggggaaa gggctggttt caggctgaca tatggccgca ggtccagttc
     2401 agcccaggtc tctcatggtt atcttccaac ccactgtcac gctgacacta tgctgccatg
     2461 cctgggctgt ggacctactg ggcatttgag gaattggaga atggagatgg caagagggca
     2521 ggcttttaag tttgggttgg agacaacttc ctgtggcccc cacaagctga gtctggcctt
     2581 ctccagctgg ccccaaaaaa ggcctttgct acatcctgat tatctctgaa agtaatcaat
     2641 caagtggctc cagtagctct ggattttctg ccagggctgg gccattgtgg tgctgcccca
     2701 gtatgacatg ggaccaaggc cagcgcaggt tatccacctc tgcctggaag tctatactct
     2761 acccagggca tccctctggt cagaggcagt gagtactggg aactggaggc tgacctgtgc
     2821 ttagaagtcc tttaatctgg gctggtacag gcctcagcct tgccctcaat gcacgaaagg
     2881 tggcccagga gagaggatca atgccatagg aggcagaagt ctggcctctg tgcctctatg
     2941 gagactatct tccagttgct gctcaacaga gttgttggct gagacctgct tgggagtctc
     3001 tgctggccct tcatctgttc aggaacacac acacacacac actcacacac gcacacacaa
     3061 tcacaatttg ctacagcaac aaaaaagaca ttgggctgtg gcattattaa ttaaagatga
     3121 tatccagtct cagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa
     3181 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg
     3241 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3301 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3361 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3421 ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa
     3481 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     3541 tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg
     3601 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3661 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3781 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3841 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3901 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3961 ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc tcgaaccaaa
     4021 ggctggagcc accatggctt ccaaggtgta cgaccccgag caacgcaaac gcatgatcac
     4081 tgggcctcag tggtgggctc gctgcaagca aatgaacgtg ctggactcct tcatcaacta
     4141 ctatgattcc gagaagcacg ccgagaacgc cgtgattttt ctgcatggta acgctgcctc
     4201 cagctacctg tggaggcacg tcgtgcctca catcgagccc gtggctagat gcatcatccc
     4261 tgatctgatc ggaatgggta agtccggcaa gagcgggaat ggctcatatc gcctcctgga
     4321 tcactacaag tacctcaccg cttggttcga gctgctgaac cttccaaaga aaatcatctt
     4381 tgtgggccac gactgggggg cttgtctggc ctttcactac tcctacgagc accaagacaa
     4441 gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg atcgagtcct gggacgagtg
     4501 gcctgacatc gaggaggata tcgccctgat caagagcgaa gagggcgaga aaatggtgct
     4561 tgagaataac ttcttcgtcg agaccatgct cccaagcaag atcatgcgga aactggagcc
     4621 tgaggagttc gctgcctacc tggagccatt caaggagaag ggcgaggtta gacggcctac
     4681 cctctcctgg cctcgcgaga tccctctcgt taagggaggc aagcccgacg tcgtccagat
     4741 tgtccgcaac tacaacgcct accttcgggc cagcgacgat ctgcctaaga tgttcatcga
     4801 gtccgaccct gggttctttt ccaacgctat tgtcgaggga gctaagaagt tccctaacac
     4861 cgagttcgtg aaggtgaagg gcctccactt cagccaggag gacgctccag atgaaatggg
     4921 taagtacatc aagagcttcg tggagcgcgt gctgaagaac gagcagaccg gtggtgggag
     4981 cggaggtggc ggatcaggtg gcggaggctc cggagggatt gaacaagatg gattgcacgc
     5041 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5101 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5161 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5221 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5281 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5341 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5401 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5461 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5521 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5581 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5641 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5701 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5761 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5821 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat
     5881 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat
     5941 cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     6001 tcagcaccgt ttctgcggac tggctttcta cgtaatggtt tcttagacgt caggtggcac
     6061 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6121 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6181 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6241 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6301 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6361 cgaagaacgt tttccaatga tgagcacttt caaagttctg ctatgtggcg cggtattatc
     6421 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6481 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6541 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaactat
     6601 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6661 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6721 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6781 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6841 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6901 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6961 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7021 ctcactgatt aagcattggt aattcgaaat gaccgaccaa gcgacgccca accggtatca
     7081 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7141 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7201 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7261 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7321 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7381 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7441 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7501 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7561 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7621 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7681 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7741 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
     7801 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7861 atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg
     7921 cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat
     7981 taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt
     8041 tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg
     8101 ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     8161 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     8221 ggcgtagagg atcgagatct accgggtagg ggaggcgctt ttcccaaggc agtctggagc
     8281 atg
//
