LOCUS       sequence-230191-        7617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of miR-124 in eurkaryotic cells, encoded in a human 
            beta-globin intron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7617)
  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 7617)
  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: pSMPUW-miR-124-GFP-Puro
FEATURES             Location/Qualifiers
     source          1..7617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        130..341
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(1582..1584,1585..1587,1588..2298)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1582..1584,1585..1587,1588..2298)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1627..1648)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1888..1907)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2235..2256
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2305..2904
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2305..2324)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2801..2821
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2917..3505
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2970..2990)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3388..3399)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3695..3875
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        3944..4154
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4013..4148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4264..4385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4355..4374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4437..4454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4608..5196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4688..4707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5448..5487)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(5497..6312)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5497..6312)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6220..6239
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6304..6323
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     enhancer        6492..6795
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6796..6999
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6950..6970
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7013..7193
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7240..7364
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7399..7515
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ccagggaccg tcgttaaact cccactaacg tagaacccag agatcgctgc gttcccgccc
       61 cctcacccgc ccgctctcgt catcactgag gtggagaaga gcatgcgtga ggctccggtg
      121 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      181 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      241 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      301 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtacacata ttgaccaaat
      361 cagggtaatt ttgcatttgt aattttaaaa aatgctttct tcttttaata tacttttttg
      421 tttatcttat ttctaatact ttccctaatc tctttctttc agggcaataa tgatacaatg
      481 tatcatgcct ctttgcacca ttctaaagaa taacagtgat aatttctggg ttaaggcaat
      541 agccgattag ttctcgagga ttcgactgat cgatccagta ccgaggagat ctgcgccgcg
      601 atcgcgcagg aaaaagcctg gatgcgaaag gatgggggag aacaaagagc ctttggaaga
      661 cgtcgctgtt atctcattgt ctgtgtgatt gggggagctg cggcggggag gatgctgtgg
      721 tcccttcctc cggcgttccc cacccccatc cctctccccg ctgtcagtgc gcacgcacac
      781 gcgccgcttt ttatttcttt ttcctggttt tcttattcca tcttctaccc acccctcttc
      841 ctttctttca cctttccttc cttccttcct cctttccttc ctcaggagaa aggcctctct
      901 ctccgtgttc acagcggacc ttgatttaaa tgtccataca attaaggcac gcggtgaatg
      961 ccaagaatgg ggctggctga gcaccgtggg tcggcgaggg cccgccaagg aaggagcgac
     1021 cgaccgagcc aggcgccctc cgcagacctc cgcgcagcgg ccgcgggcgc gaggggaggg
     1081 gtctggagct ccctccggct gcctgtcccg caccggagcc cgtggggtgg ggaggtgtgc
     1141 agcctgtgac agacaggggc ttagagatgc aaacagactc agggagagaa acagaagctg
     1201 attctgtgac agaagcagat ctgtgcagca cagatgcggt gtgcgtgggg aacgcgtacg
     1261 cggccgctcg agcagaaact catctcagat aagtcgctag ctcgagcttt tggagaatat
     1321 ttctgcatat aaatatttct gcatataaat tgtaactgat gtaagaggtt tcatattgct
     1381 aatagcagct acaatccagc taccattctg cttttatttt atggttggga taaggctgga
     1441 ttattctgag tccaagctag gcccttttgc taatcatgtt catacctctt atcttcctcc
     1501 cacagctcct gggcaacgtg ctggtctgtg tgctggccca tcactttggc aaagcacgtg
     1561 agatctgaat tctgacacac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1621 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1681 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1741 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1801 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1861 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1921 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1981 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2041 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2101 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2161 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2221 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2281 atggacgagc tgtacaaggc caccatgacc gagtacaagc ccacggtgcg cctcgccacc
     2341 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     2401 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     2461 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     2521 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     2581 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     2641 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     2701 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     2761 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     2821 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     2881 atgacccgca agcccggtgc ctgagcggcc gtcgacaatc aacctctgga ttacaaaatt
     2941 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3001 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3061 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3121 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3181 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3241 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3301 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     3361 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3421 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3481 atctcccttt gggccgcctc cccgcttagt actggtacct ttaagaccaa tgacttacaa
     3541 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     3601 ctcccaacga agacaagatt ccggaattta tttgtgaaat ttgtgatgct attgctttat
     3661 ttgtaaaccg gtgcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
     3721 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     3781 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     3841 cagacccttt tagtcagtgt ggaaaatctc tagcatctag agtatgcaaa gcatgcatct
     3901 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4021 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4081 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4141 tggaggccta ggctagagat cataatcagc cataccacat ttgtagaggt tttacttgct
     4201 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4261 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4321 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4381 tcttatcatg tctgctagca gagcgggctt ttttttctta ggccttcttc cgcttcctcg
     4441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc tgcgcagtcc aaaaaaaaag
     5461 gctccaaaag gagcctttaa ttgtatcggt gggcccttag aaaaactcat cgagcatcaa
     5521 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     5581 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     5641 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     5701 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     5761 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     5821 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     5881 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     5941 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     6001 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     6061 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     6121 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     6181 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     6241 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     6301 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     6361 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggtt
     6421 taaacaaata gtcaaaagcc tccgggcgac tagtcggggt cattagttca tagcccatat
     6481 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6541 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6601 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6661 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6721 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     6781 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     6841 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     6901 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     6961 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgggtctct
     7021 ctggttagac cagatttgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     7081 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     7141 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
     7201 ccgaacaggg acctgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
     7261 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tgcagagtac gccaaaattt
     7321 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     7381 aaatagcggc cgccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     7441 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     7501 aaaaattcaa attttcgggg gatcttcgat gcctccccgt caccaccccc cccaacccgc
     7561 cccgaccgga gctgagagta attcatacaa aaggactcgc ccctgccttg gggaatc
//
