LOCUS       sequence-230212-        7521 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Dual luciferase assay negative control for miR-124 binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7521)
  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 7521)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-miR-124-sponge_rev(neg. CTRL)
FEATURES             Location/Qualifiers
     source          1..7521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..517
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(53..76)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     450..469
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      558..567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             564..2216
                     /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    2555..2676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2566..2585)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2620..2639
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2921..3278
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2921..3112
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2944..2964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3129..3264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3323..3332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3329..4261
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4313..5104
                     /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(4364..4383)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4974..4993
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5168..5216
                     /note="synthetic polyadenylation signal"
     promoter        5372..5476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5477..5545,5546..6337)
                     /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(5695..5714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6495..7083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6575..6594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7199..7482
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 gaggatcgag atctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
       61 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
      121 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
      181 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
      241 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
      301 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
      361 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
      421 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
      481 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttggcaat
      541 ccggtactgt tggtaaagcc accatggaag atgccaaaaa cattaagaag ggcccagcgc
      601 cattctaccc actcgaagac gggaccgccg gcgagcagct gcacaaagcc atgaagcgct
      661 acgccctggt gcccggcacc atcgccttta ccgacgcaca tatcgaggtg gacattacct
      721 acgccgagta cttcgagatg agcgttcggc tggcagaagc tatgaagcgc tatgggctga
      781 atacaaacca tcggatcgtg gtgtgcagcg agaatagctt gcagttcttc atgcccgtgt
      841 tgggtgccct gttcatcggt gtggctgtgg ccccagctaa cgacatctac aacgagcgcg
      901 agctgctgaa cagcatgggc atcagccagc ccaccgtcgt attcgtgagc aagaaagggc
      961 tgcaaaagat cctcaacgtg caaaagaagc taccgatcat acaaaagatc atcatcatgg
     1021 atagcaagac cgactaccag ggcttccaaa gcatgtacac cttcgtgact tcccatttgc
     1081 cacccggctt caacgagtac gacttcgtgc ccgagagctt cgaccgggac aaaaccatcg
     1141 ccctgatcat gaacagtagt ggcagtaccg gattgcccaa gggcgtagcc ctaccgcacc
     1201 gcaccgcttg tgtccgattc agtcatgccc gcgaccccat cttcggcaac cagatcatcc
     1261 ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg cttcggcatg ttcaccacgc
     1321 tgggctactt gatctgcggc tttcgggtcg tgctcatgta ccgcttcgag gaggagctat
     1381 tcttgcgcag cttgcaagac tataagattc aatctgccct gctggtgccc acactattta
     1441 gcttcttcgc taagagcact ctcatcgaca agtacgacct aagcaacttg cacgagatcg
     1501 ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc cgtggccaaa cgcttccacc
     1561 taccaggcat ccgccagggc tacggcctga cagaaacaac cagcgccatt ctgatcaccc
     1621 ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt gcccttcttc gaggctaagg
     1681 tggtggactt ggacaccggt aagacactgg gtgtgaacca gcgcggcgag ctgtgcgtcc
     1741 gtggccccat gatcatgagc ggctacgtta acaaccccga ggctacaaac gctctcatcg
     1801 acaaggacgg ctggctgcac agcggcgaca tcgcctactg ggacgaggac gagcacttct
     1861 tcatcgtgga ccggctgaag agcctgatca aatacaaggg ctaccaggta gccccagccg
     1921 aactggagag catcctgctg caacacccca acatcttcga cgccggggtc gccggcctgc
     1981 ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac ggtaaaacca
     2041 tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc aagaagctgc
     2101 gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag ttggacgccc
     2161 gcaagatccg cgagattctc attaaggcca agaagggcgg caagatcgcc gtgtaattct
     2221 agttgtttaa acgagtccct cgacgaattc ctactgtcga ctaaggcaca tttgaatgcc
     2281 atttgatttt atttacgaaa taaggcacct ttgaatgcca ttgattggag ttttggagtt
     2341 aaggcacttt tgaatgccat tttggattag aaaggattta aggcaccgtt gaatgcctct
     2401 agacatatca gtgacggttc catcggagtc gacctgcagg catgcaagct gatccggctg
     2461 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     2521 aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2581 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2641 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2701 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     2761 tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     2821 ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc
     2881 ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc
     2941 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3001 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3061 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3121 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3181 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3241 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa
     3301 cagtctcgaa ccaaaggctg gagccaccat ggcttccaag gtgtacgacc ccgagcaacg
     3361 caaacgcatg atcactgggc ctcagtggtg ggctcgctgc aagcaaatga acgtgctgga
     3421 ctccttcatc aactactatg attccgagaa gcacgccgag aacgccgtga tttttctgca
     3481 tggtaacgct gcctccagct acctgtggag gcacgtcgtg cctcacatcg agcccgtggc
     3541 tagatgcatc atccctgatc tgatcggaat gggtaagtcc ggcaagagcg ggaatggctc
     3601 atatcgcctc ctggatcact acaagtacct caccgcttgg ttcgagctgc tgaaccttcc
     3661 aaagaaaatc atctttgtgg gccacgactg gggggcttgt ctggcctttc actactccta
     3721 cgagcaccaa gacaagatca aggccatcgt ccatgctgag agtgtcgtgg acgtgatcga
     3781 gtcctgggac gagtggcctg acatcgagga ggatatcgcc ctgatcaaga gcgaagaggg
     3841 cgagaaaatg gtgcttgaga ataacttctt cgtcgagacc atgctcccaa gcaagatcat
     3901 gcggaaactg gagcctgagg agttcgctgc ctacctggag ccattcaagg agaagggcga
     3961 ggttagacgg cctaccctct cctggcctcg cgagatccct ctcgttaagg gaggcaagcc
     4021 cgacgtcgtc cagattgtcc gcaactacaa cgcctacctt cgggccagcg acgatctgcc
     4081 taagatgttc atcgagtccg accctgggtt cttttccaac gctattgtcg agggagctaa
     4141 gaagttccct aacaccgagt tcgtgaaggt gaagggcctc cacttcagcc aggaggacgc
     4201 tccagatgaa atgggtaagt acatcaagag cttcgtggag cgcgtgctga agaacgagca
     4261 gaccggtggt gggagcggag gtggcggatc aggtggcgga ggctccggag ggattgaaca
     4321 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4381 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4441 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4501 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4561 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4621 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4681 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4741 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4801 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4861 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4921 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4981 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5041 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5101 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5161 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc
     5221 gatagcgata aggatcctct ttgcgcttgc gttttccctt gtccagatag cccagtagct
     5281 gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtaa tggtttctta
     5341 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5401 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5461 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5521 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5581 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5641 tgagagtttt cgccccgaag aacgttttcc aatgatgagc actttcaaag ttctgctatg
     5701 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5761 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5821 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5881 acttctgaca actatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5941 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6001 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6061 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6121 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6181 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6241 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6301 cgctgagata ggtgcctcac tgattaagca ttggtaattc gaaatgaccg accaagcgac
     6361 gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6901 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt
     7081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc
     7201 ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc atgaaaaatg
     7261 gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg cataggtatt
     7321 catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt
     7381 tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta caattaaggg
     7441 attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc
     7501 ggccacgatg cgtccggcgt a
//
