LOCUS       sequence-232239-        8084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockdown of miR-125.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8084)
  AUTHORS   Akerblom M, Petri R, Sachdeva R, Klussendorf T, Mattsson B, Gentner 
            B, Jakobsson J
  TITLE     microRNA-125 distinguishes developmentally generated and adult-born 
            olfactory bulb interneurons.
  JOURNAL   Development. 2014 Apr;141(7):1580-8. doi: 10.1242/dev.101659. Epub 
            2014 Mar 5.
  PUBMED    24598163
REFERENCE   2  (bases 1 to 8084)
  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: 
            "10.1242/dev.101659"; journalName: "Development"; date: "2014-04"; 
            volume: "141"; issue: "7"; pages: "1580-8"
COMMENT     Sequence Label: pLV.CMV.GFPmiR125sponge
FEATURES             Location/Qualifiers
     source          1..8084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     624..643
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        624..642
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      663..798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    817..938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(828..847)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     882..901
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             1010..1243
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             1594..1605
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2003..2023
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2092..2805,2806..2808,2809..2811))
                     /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             complement(join(2095..2805,2806..2808,2809..2811))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2137..2158)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2486..2505
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2745..2766
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2808..2817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2907..3110)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        3111..3414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    3437..3554
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(3888..3932)
                     /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"
     misc_feature    4117..4350
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    4843..4968
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5015..5195
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(5210..5413)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        5414..5793
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5894..5914)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5894..5912)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5933..5949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5933..5949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5946..5968)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5957..5973
                     /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(5981..5987,5988..6005,6006..6011))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6026..6047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6164..6181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6335..6923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6415..6434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7094..7885,7886..7954))
                     /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     7717..7736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7955..8059)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtacgtccc
      661 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
      721 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
      781 cggagttagg ggcgggatag ctagagccag acatgataag atacattgat gagtttggac
      841 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
      901 ctttatttgt aaccattata agctgcaata aacaagttcc tctcactctc tgatattcat
      961 ttctttgcaa gttataaata ctgaataata agatgacatg aactactact gctagagatt
     1021 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
     1081 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
     1141 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacaagca
     1201 aaaagcagat cttgtcttcg ttgggagtga attagccctt ccagtccccc cttttctttt
     1261 aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg tacctacgtc
     1321 acaagtttag tctcagggat acgtcacaag tttagtctca gggatacgtc acaagtttag
     1381 tctcagggat acgtcacaag tttagtctca gggatacgtc acaagtttag tctcagggat
     1441 acgtcacaag tttagtctca gggatacgtc acaagtttag tctcagggat acgtcacaag
     1501 tttagtctca gggatatcta gaatcccggg aaggcgaaga cgcggaagag gctctagagc
     1561 cggcagcagg ccgcgggaag gaaggtccgc tggattgagg gccgaaggga cgtagcagaa
     1621 ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc agccatggaa aggacgtcag
     1681 cttccccgac aacaccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg
     1741 ggcaaggcag gcggcgatga gttccgccgt ggcaataggg agggggaaag cgaaagtccc
     1801 ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac
     1861 agtgcacacc acgccacgtt gcctgacaac gggccacaac tcctcataaa gagacagcaa
     1921 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     1981 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     2041 caatctttca caaattttgt aatccagagg ttgattgtcg acgcggccgc tttacttgta
     2101 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
     2161 atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc
     2221 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg
     2281 cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg
     2341 cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag
     2401 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
     2461 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
     2521 gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
     2581 gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca
     2641 gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc
     2701 atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
     2761 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac
     2821 cggtggatcc tctagagtcg gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc
     2881 caggcgatct gacggttcac taaacgagct ctgcttatat agacctccca ccgtacacgc
     2941 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     3001 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     3061 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     3121 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     3181 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     3241 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     3301 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     3361 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     3421 tcccaagctt atcgataaaa ttttgaattt ttgtaatttg tttttgtaat tctttagttt
     3481 gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc
     3541 cttttctttt aaaagttaac cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3601 gcctccatcc agtctattaa ttgttgccga taccgtcgag atccgttcac taatcgaatg
     3661 gatctgtctc tgtctctctc tccaccttct tcttctattc cttcgggcct gtcgggtccc
     3721 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     3781 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     3841 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     3901 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     3961 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     4021 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     4081 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     4141 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     4201 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     4261 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     4321 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     4381 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     4441 ttatataatt cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagc
     4501 ggccgcttgc tgtgcggtgg tcttactttt gttttgctct tcctctatct tgtctaaagc
     4561 ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt
     4621 atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat
     4681 ttgtctacag ccttctgatg tttctaacag gccaggatta actgcgaatc gttctagctc
     4741 cctgcttgcc catactatat gttttaattt atattttttc tttccccctg gccttaaccg
     4801 aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc
     4861 tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc
     4921 gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt
     4981 ccctttcgct ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa
     5041 aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga
     5101 agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc
     5161 tcccaggctc agatctggtc taaccagaga gaccccggtt cactaaacga gctctgctta
     5221 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     5281 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     5341 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     5401 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     5461 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     5521 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     5581 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     5641 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     5701 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     5761 attactatta ataactagtc aataatcaat gtcaacatgg cggtaatgtt ggacatgagc
     5821 caatataaat gtacatatta tgatatggat acaacgtatg caatggccaa gcttgcagct
     5881 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     5941 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6001 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6061 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6121 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6181 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6241 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6301 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6361 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6421 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6481 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6541 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6601 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6661 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6721 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6781 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6841 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6901 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6961 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7021 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7081 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7141 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7201 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7261 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7321 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7381 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7441 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7501 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7561 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7621 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7681 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7741 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7801 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7861 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7921 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7981 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8041 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acct
//
