LOCUS       sequence-224558-        8094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kockdown of miR-124.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8094)
  AUTHORS   Akerblom M, Sachdeva R, Barde I, Verp S, Gentner B, Trono D, 
            Jakobsson J
  TITLE     MicroRNA-124 is a subventricular zone neuronal fate determinant.
  JOURNAL   J Neurosci. 2012 Jun 27;32(26):8879-89. doi: 
            10.1523/JNEUROSCI.0558-12.2012.
  PUBMED    22745489
REFERENCE   2  (bases 1 to 8094)
  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: 
            "10.1523/JNEUROSCI.0558-12"; journalName: "J Neurosci"; date: 
            "2012-06-27- 27"; volume: "32"; issue: "26"; pages: "8879-89"
COMMENT     Sequence Label: pLV.CMV.GFP.miR124.sponge
FEATURES             Location/Qualifiers
     source          1..8094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    60..177
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        200..503
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        504..707
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      797..806
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(803..805,806..808,809..1522)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(803..805,806..808,809..1519)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(848..869)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1109..1128)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1456..1477
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1591..1611)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2009..2020)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2381..2614
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2686..2807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2723..2742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2777..2796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2826..2961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2981..3000)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2982..3000)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3010..3027)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3010..3026)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3019..3041)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3168..3623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3255..3274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3465..3486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3649..3753
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3754..3822,3823..4614)
                     /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(3972..3991)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4785..5373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5274..5293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5527..5544
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5661..5682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5697..5702,5703..5720,5721..5727)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5735..5751
                     /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)."
     primer_bind     5740..5762
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5759..5775
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5759..5775
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5794..5814
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5796..5814
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        5915..6294
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6295..6498
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6513..6693
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6740..6865
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7358..7591
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7776..7820
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggttaact
       61 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
      121 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc
      181 gataagcttg ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      241 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      361 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      421 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      481 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      541 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      601 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      661 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
      721 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgactct
      781 agaggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      841 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      901 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      961 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1021 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1081 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1141 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1201 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1261 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1321 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1381 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1441 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1501 catggacgag ctgtacaagt aaagcggccg cgtcgacaat caacctctgg attacaaaat
     1561 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1621 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     1681 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     1741 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     1801 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     1861 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     1921 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     1981 gcgcgggacg cccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2041 cggcctgctg ccggctctag agtggcattc atacgtgcct taacgtatgg cattcatacg
     2101 tgccttaagc tagagtggca ttcatacgtg ccttaacgta tggcattcat acgtgcctta
     2161 agctagagtg gcattcatac gtgccttaac gtatggcatt catacgtgcc ttaagctaga
     2221 gtggcattca tacgtgcctt aacgtatggc attcatacgt gccttaagct agagcctctt
     2281 ccgcgtcttc gccttcccgg gtcgagctcg gtacctttaa gaccaatgac ttacaaggca
     2341 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     2401 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     2461 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     2521 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     2581 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     2641 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     2701 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2761 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     2821 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2881 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2941 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     3001 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3061 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3121 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     3181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3241 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3301 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3361 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3421 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3481 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3541 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3601 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     3661 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3721 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3781 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3841 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3901 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3961 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4021 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4081 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4141 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4201 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4261 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4321 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4381 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4441 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4501 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4561 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4621 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4681 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4741 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4801 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4861 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4921 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4981 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5041 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5101 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5161 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5221 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5281 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5341 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5401 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5461 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5521 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     5581 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     5641 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     5701 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     5761 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     5821 aagctggagc tgcaagcttg gccattgcat acgttgtatc catatcataa tatgtacatt
     5881 tatattggct catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa
     5941 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6001 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6061 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6121 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     6181 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     6241 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     6301 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     6361 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     6421 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     6481 gtctatataa gcagagctcg tttagtgaac cggggtctct ctggttagac cagatctgag
     6541 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6601 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6661 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     6721 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     6781 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     6841 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     6901 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     6961 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     7021 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     7081 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     7141 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     7201 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     7261 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     7321 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     7381 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     7441 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     7501 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     7561 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     7621 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     7681 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     7741 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     7801 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     7861 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     7921 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     7981 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     8041 acagatccat tcgattagtg aacggatctc gacggtatcg gcaacaatta atag
//
