LOCUS       sequence-231766-        7989 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 7989)
  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 7989)
  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-ZNF787-203
FEATURES             Location/Qualifiers
     source          1..7989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..547
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(83..106)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     480..499
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      588..597
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             594..2246
                     /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    3053..3174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3064..3083)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3118..3137
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3419..3776
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3419..3610
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3442..3462)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3627..3762
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3821..3830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3827..4759
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4811..5602
                     /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(4862..4881)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5472..5491
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5666..5714
                     /note="synthetic polyadenylation signal"
     promoter        5870..5974
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5975..6043,6044..6835)
                     /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(6193..6212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6993..7581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7073..7092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7697..7980
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 cagtactccg gccacgatgc gtccggcgta gaggatcgag atctaccggg taggggaggc
       61 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
      121 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
      181 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
      241 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
      301 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
      361 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
      421 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
      481 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
      541 cctttcgacc tgcagcccaa gcttggcaat ccggtactgt tggtaaagcc accatggaag
      601 atgccaaaaa cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg
      661 gcgagcagct gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta
      721 ccgacgcaca tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc
      781 tggcagaagc tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg
      841 agaatagctt gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg
      901 ccccagctaa cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc
      961 ccaccgtcgt attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc
     1021 taccgatcat acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa
     1081 gcatgtacac cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc
     1141 ccgagagctt cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg
     1201 gattgcccaa gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc
     1261 gcgaccccat cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat
     1321 ttcaccacgg cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg
     1381 tgctcatgta ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc
     1441 aatctgccct gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca
     1501 agtacgacct aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg
     1561 taggtgaggc cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga
     1621 cagaaacaac cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag
     1681 gcaaggtggt gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg
     1741 gtgtgaacca gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta
     1801 acaaccccga ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca
     1861 tcgcctactg ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca
     1921 aatacaaggg ctaccaggta gccccagccg aactggagag catcctgctg caacacccca
     1981 acatcttcga cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg
     2041 cagtcgtcgt gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg
     2101 ccagccaggt tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc
     2161 ctaaaggact gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca
     2221 agaagggcgg caagatcgcc gtgtaattct agttgtttaa acgagctcgg ggaggggccc
     2281 gggggggggc gggcagggcc cctcgggctt ggcgggagcg acgacgtgca agaccaagag
     2341 acccagggag aagcggacgg ggtgcgatgg accccgcggc atccccggct ccccgccctc
     2401 ctctgtccct gcgccgtccg tgggggggca ctggggcgta ttagggcccg ttcacttctc
     2461 cgcctcgaga ctctgccccc tcagcctggc ccgcccttcc taagttatcg gcctgcgagc
     2521 cggagagaca atggagagaa ctgcagaggc cgagtggcga ccaggcccca agcggaaggc
     2581 cattttccct gcccttcctc agttgtccgg ggcgggggtg gggggtgctg cgaggaatgg
     2641 agaaaccagg ccctttagag gatcccagag agggaagaga accaggggcc cttttctctt
     2701 ttagaatttt cttttaatca gcccaccttt tgactccccg cccgccccaa tccattattt
     2761 ttttctgcct tccgggtccc atccttagag ctcgagtcgt tccttcccct cctggattcc
     2821 gggtagcaga ggcagcgccg caggaggcgg gtgcccgttt gtcccaggcc tgggcgtggt
     2881 ggaggaaggg gtgctggaac aataaacggc actatccaaa tgtcagtcga cctgcaggca
     2941 tgcaagctga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     3001 ctgagcaata actagcataa ccccttgggg cggccgcttc gagcagacat gataagatac
     3061 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3121 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3181 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     3241 aagtaaaacc tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat
     3301 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga
     3361 tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct
     3421 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3481 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3541 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3601 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3661 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3721 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3781 ttgattcttc tgacacaaca gtctcgaacc aaaggctgga gccaccatgg cttccaaggt
     3841 gtacgacccc gagcaacgca aacgcatgat cactgggcct cagtggtggg ctcgctgcaa
     3901 gcaaatgaac gtgctggact ccttcatcaa ctactatgat tccgagaagc acgccgagaa
     3961 cgccgtgatt tttctgcatg gtaacgctgc ctccagctac ctgtggaggc acgtcgtgcc
     4021 tcacatcgag cccgtggcta gatgcatcat ccctgatctg atcggaatgg gtaagtccgg
     4081 caagagcggg aatggctcat atcgcctcct ggatcactac aagtacctca ccgcttggtt
     4141 cgagctgctg aaccttccaa agaaaatcat ctttgtgggc cacgactggg gggcttgtct
     4201 ggcctttcac tactcctacg agcaccaaga caagatcaag gccatcgtcc atgctgagag
     4261 tgtcgtggac gtgatcgagt cctgggacga gtggcctgac atcgaggagg atatcgccct
     4321 gatcaagagc gaagagggcg agaaaatggt gcttgagaat aacttcttcg tcgagaccat
     4381 gctcccaagc aagatcatgc ggaaactgga gcctgaggag ttcgctgcct acctggagcc
     4441 attcaaggag aagggcgagg ttagacggcc taccctctcc tggcctcgcg agatccctct
     4501 cgttaaggga ggcaagcccg acgtcgtcca gattgtccgc aactacaacg cctaccttcg
     4561 ggccagcgac gatctgccta agatgttcat cgagtccgac cctgggttct tttccaacgc
     4621 tattgtcgag ggagctaaga agttccctaa caccgagttc gtgaaggtga agggcctcca
     4681 cttcagccag gaggacgctc cagatgaaat gggtaagtac atcaagagct tcgtggagcg
     4741 cgtgctgaag aacgagcaga ccggtggtgg gagcggaggt ggcggatcag gtggcggagg
     4801 ctccggaggg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4861 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4921 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4981 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5041 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5101 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5161 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5221 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5281 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5341 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5401 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5461 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5521 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5581 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5641 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5701 tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt
     5761 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     5821 ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5881 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5941 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6001 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6061 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6121 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6181 tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6241 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6301 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6361 taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc taaccgcttt
     6421 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6481 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6541 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6601 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6661 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6721 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6781 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaattcga
     6841 aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta
     6901 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7561 cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7681 tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg
     7741 gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc
     7801 tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc
     7861 gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc
     7921 ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga
     7981 gataaaccg
//
