LOCUS       sequence-231695-        8332 bp    DNA     circular SYN 15-APR-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 8332)
  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 8332)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SUMF1-201
FEATURES             Location/Qualifiers
     source          1..8332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..519
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(55..78)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     452..471
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      560..569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             566..2218
                     /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    3368..3489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3379..3398)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3433..3452
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3734..4091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3734..3925
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3757..3777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3942..4077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4136..4145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4142..5074
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             5126..5917
                     /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(5177..5196)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5787..5806
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5981..6029
                     /note="synthetic polyadenylation signal"
     promoter        6185..6289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6290..6358,6359..7150)
                     /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(6508..6527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7308..7896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7388..7407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8012..8295
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tagaggatcg agatctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
       61 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
      121 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
      181 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
      241 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
      301 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
      361 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
      421 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
      481 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttggca
      541 atccggtact gttggtaaag ccaccatgga agatgccaaa aacattaaga agggcccagc
      601 gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg
      661 ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac
      721 ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct
      781 gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt
      841 gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg
      901 cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg
      961 gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat
     1021 ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt
     1081 gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat
     1141 cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca
     1201 ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat
     1261 ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac
     1321 gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct
     1381 attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt
     1441 tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat
     1501 cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca
     1561 cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac
     1621 ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa
     1681 ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt
     1741 ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat
     1801 cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt
     1861 cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc
     1921 cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct
     1981 gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac
     2041 catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct
     2101 gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc
     2161 ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgtaatt
     2221 ctagttgttt aaacgagctc gcaaccaagg aaagtcttcc ccagtccaag gagcagtcgt
     2281 gtctgaccta cattgggctt ttctcagaac tttgaacgat cccatgcaaa gaattcccac
     2341 cctgaggtgg gttacatacc tgcccaatgg ccaaaggaac cgccttgtga gaccaaattg
     2401 ctgacctggg tcagtgcatg tgctttatgg tgtggtgcat ctttggagat catcgccata
     2461 ttttactttt gagagtcttt aaagaggaag gggagtggag ggaaccctga gctaggcttc
     2521 aggaggcccg cgtcctacgc aggctctgcc acaggggtta gaccccaggt ccgacgcttg
     2581 accttcctgg gcctcaagtg ccctccccta tcaaatgaag ggatggacag catgacctct
     2641 gggtgtctct ccaactcacc agttctaaaa agggtatcag attctattgt gacttcatag
     2701 tgagaattta tgatagatta ttttttagct attttttcca tgtgtgaacc ttgagtgata
     2761 ctaatcatgt aaagtaagag ttctcttatg tattattttc ggaagagggg tgtggtgact
     2821 cctttatatt cgtactgcac tttgtttttc caaggaaatc agtgtctttt acgttgttat
     2881 gatgaatccc acatggggcc ggtgatggta tgctgcagtt cagccgttga acacatagga
     2941 atgtctgtgg ggtgactcta ctgtgcttta tcttttaaca ttaagtgcct ttggttcaga
     3001 ggggcagtca taagctctgt ttccccctct ccccaaagcc ttcagcgaac gtgaaatgtg
     3061 cgctaaacgg ggaaacctgt ttaattctag atatagggaa aaaggaacga ggaccttgaa
     3121 tgagctatat tcagggtatc cggtattttg taatagggaa taggaaacct tgttggctgt
     3181 ggaatatccg atgctttgaa tcatgcactg tgttgaataa acgtatctgc taaatcagga
     3241 gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt
     3301 tggctgctgc caccgctgag caataactag cataacccct tggggcggcc gcttcgagca
     3361 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3421 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3481 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3541 gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt ttaacaaaat
     3601 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3661 caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
     3721 ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3841 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3901 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3961 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4021 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4081 gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaaccaaagg ctggagccac
     4141 catggcttcc aaggtgtacg accccgagca acgcaaacgc atgatcactg ggcctcagtg
     4201 gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc atcaactact atgattccga
     4261 gaagcacgcc gagaacgccg tgatttttct gcatggtaac gctgcctcca gctacctgtg
     4321 gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc atcatccctg atctgatcgg
     4381 aatgggtaag tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc actacaagta
     4441 cctcaccgct tggttcgagc tgctgaacct tccaaagaaa atcatctttg tgggccacga
     4501 ctggggggct tgtctggcct ttcactactc ctacgagcac caagacaaga tcaaggccat
     4561 cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc ctgacatcga
     4621 ggaggatatc gccctgatca agagcgaaga gggcgagaaa atggtgcttg agaataactt
     4681 cttcgtcgag accatgctcc caagcaagat catgcggaaa ctggagcctg aggagttcgc
     4741 tgcctacctg gagccattca aggagaaggg cgaggttaga cggcctaccc tctcctggcc
     4801 tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc gtccagattg tccgcaacta
     4861 caacgcctac cttcgggcca gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg
     4921 gttcttttcc aacgctattg tcgagggagc taagaagttc cctaacaccg agttcgtgaa
     4981 ggtgaagggc ctccacttca gccaggagga cgctccagat gaaatgggta agtacatcaa
     5041 gagcttcgtg gagcgcgtgc tgaagaacga gcagaccggt ggtgggagcg gaggtggcgg
     5101 atcaggtggc ggaggctccg gagggattga acaagatgga ttgcacgcag gttctccggc
     5161 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5221 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5281 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5341 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5401 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5461 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5521 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5581 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5641 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5701 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5761 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5821 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5881 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5941 accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc
     6001 attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct
     6061 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     6121 ctgcggactg gctttctacg taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     6181 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6241 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6301 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6361 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6421 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6481 tccaatgatg agcactttca aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6541 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6601 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6661 cataaccatg agtgataaca ctgcggccaa cttacttctg acaactatcg gaggaccgaa
     6721 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6781 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6841 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6901 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6961 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7021 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7081 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7141 gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag
     7201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7261 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7321 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7381 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7441 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7501 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7561 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7621 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7681 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7741 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7801 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7861 aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg
     7921 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7981 aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg
     8041 cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta
     8101 atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga
     8161 ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc
     8221 tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc
     8281 cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg cg
//
