LOCUS       sequence-354354-        6036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The pre-miR-124 sequence has been inserted into the EcoRV of 
            pCAG-nDsRedFL-intron..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6036)
  AUTHORS   Suzuki F, Okuno M, Tanaka T, Sanuki R
  TITLE     Overexpression of neural miRNAs miR-9/9* and miR-124 suppresses 
            differentiation to Muller glia and promotes differentiation to 
            neurons in mouse retina in vivo.
  JOURNAL   Genes Cells. 2020 Nov;25(11):741-752. doi: 10.1111/gtc.12809. Epub 
            2020 Oct 21.
  PUBMED    32979863
REFERENCE   2  (bases 1 to 6036)
  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"; doi: "10.1111/gtc.12809";
            journalName: "Genes Cells"; date: "2020-11"; volume: "25"; issue: 
            "11"; pages: "741-752"
COMMENT     Sequence Label: pCAG-nDsRedFL-miR124
FEATURES             Location/Qualifiers
     source          1..6036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          647..1663
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1587..1609
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1671..1690
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1727..1736
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1739..1759
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1943..1963)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2897..2920
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2986..3051
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3067..3086)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3132..3187
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3186..3205)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3548..3564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3548..3564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3561..3583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3572..3588
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3596..3602,3603..3620,3621..3626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3641..3662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3721..3916
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3767..3902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3922..4056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3959..3978)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4013..4032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4124..4141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4295..4883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4375..4394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5054..5845,5846..5914))
                     /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     5677..5696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5915..6019)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccacctgg gtcgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      421 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      481 ttttgtgcag cgatgggggc gggggggggg ggggggcgmg gggcggggsg gggsgrggsg
      541 grgrggkgsg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
      601 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
      661 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      721 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      781 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      841 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      901 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      961 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
     1021 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
     1081 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
     1141 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
     1201 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
     1261 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
     1321 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     1381 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1441 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1501 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1561 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1621 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1681 ggttattgtg ctgtctcatc attttggcaa agaattcacc cgggaagcca ccatggctcc
     1741 aaagaagaag cgtaaggtac ggtacgcctc ctccgagaac gtcatcaccg agttcatgcg
     1801 cttcaaggtg cgcatggagg gcaccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1861 cgagggccgc ccctacgagg gccacaacac cgtgaagctg aaggtgacca agggcggccc
     1921 cctgcccttc gcctgggaca tcctgtcccc ccagttccag tacggctcca aggtgtacgt
     1981 gaagcacccc gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg
     2041 ggagcgcgtg atgaacttcg aggacggcgg cgtggcgacc gtgacccagg actcctccct
     2101 gcaggacggc tgcttcatct acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg
     2161 ccccgtgatg cagaagaaaa ctatgggctg ggaggcctcc accgagcgcc tgtacccccg
     2221 cgacggcgtg ctgaagggcg agacccacaa ggccctgaag ctgaaggacg gcggccacta
     2281 cctggtggag ttcaagtcca tctacatggc caagaagccc gtgcaggtga gtactccctc
     2341 tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg
     2401 atattcacct ggcccgcggt gatgcctttg agggatctag taggtgggag tactgctcag
     2461 agctacaact ctaggagtag ggactccaag cctagagctc caagagaggg tgaagggcag
     2521 ggagaaaatt atagtaatag ttgcaatgag tcacttgctt ctagatcaag atcagagact
     2581 ctgctctccg tgttcacagc ggaccttgat ttaatgtcat acaattaagg cacgcggtga
     2641 atgccaagag cggagcctac agctgcactt gaaggacatc cgagagaagt taggaagggt
     2701 ggggagaaac aattctagaa tgaacccatc ctgtgcgaca cgatcgtggc cgcgtccatc
     2761 tggtcagaaa agacaatctt tttgttgtca acgttgaggt gtgacaggct tgagagctgg
     2821 ccatacactt gagtgacaat gacatccact ttgcctttct ctccacagct gcccggctac
     2881 tactacgtgg acgccaagct ggacatcacc tcccacaacg aggactacac catcgtggag
     2941 cagtacgagc gcaccgaggg ccgccaccac ctgttcctgg gtaccgacta taaggaccac
     3001 gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gtaactcgag
     3061 tgtagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     3121 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3181 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     3241 ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata
     3301 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     3361 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     3421 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     3481 agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt
     3541 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3601 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3661 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
     3721 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3781 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3841 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3901 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     3961 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4021 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     4081 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4141 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4201 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4261 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4321 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4381 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4441 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4501 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4561 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4621 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4681 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4741 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4801 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4861 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4921 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4981 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5041 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5101 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5161 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5221 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5281 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5341 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5401 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5461 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5521 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5581 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5641 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5701 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5761 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5821 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5881 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5941 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6001 aacaaatagg ggttccgcgc acatttcccc gaaaag
//
