LOCUS       sequence-237376-        8493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMAGIC R4-R3 entry plasmid, contains 2x NLS Sa-dCas9 fused to the 
            LSD1 H3K4me1/2 demethylase for 3 or 4-component MultiSite Gateway 
            Pro assembly..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8493)
  AUTHORS   Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, 
            Newgard CB
  TITLE     Creation of versatile cloning platforms for transgene expression and
            dCas9-based epigenome editing.
  JOURNAL   Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 
            10.1093/nar/gky1286.
  PUBMED    30590691
REFERENCE   2  (bases 1 to 8493)
  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: "Nucleic 
            Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286."
COMMENT     Sequence Label: IF311: pMAGIC (R4-R3) NLS-Sa dCas9-NLS-LSD1
FEATURES             Location/Qualifiers
     source          1..8493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(603..726)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      726..735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             738..758
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3939..3986
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    6556..6679
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6734..6753)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6735..6753)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6758..6774)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6758..6774)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6876..6895)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6887..7696
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6954..6973)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7843..8431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8332..8351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttgccac catggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
      781 ccaagcggaa ctacatcctg ggcctggcca tcggcatcac cagcgtgggc tacggcatca
      841 tcgactacga gacacgggac gtgatcgatg ccggcgtgcg gctgttcaaa gaggccaacg
      901 tggaaaacaa cgagggcagg cggagcaaga gaggcgccag aaggctgaag cggcggaggc
      961 ggcatagaat ccagagagtg aagaagctgc tgttcgacta caacctgctg accgaccaca
     1021 gcgagctgag cggcatcaac ccctacgagg ccagagtgaa gggcctgagc cagaagctga
     1081 gcgaggaaga gttctctgcc gccctgctgc acctggccaa gagaagaggc gtgcacaacg
     1141 tgaacgaggt ggaagaggac accggcaacg agctgtccac caaagagcag atcagccgga
     1201 acagcaaggc cctggaagag aaatacgtgg ccgaactgca gctggaacgg ctgaagaaag
     1261 acggcgaagt gcggggcagc atcaacagat tcaagaccag cgactacgtg aaagaagcca
     1321 aacagctgct gaaggtgcag aaggcctacc accagctgga ccagagcttc atcgacacct
     1381 acatcgacct gctggaaacc cggcggacct actatgaggg acctggcgag ggcagcccct
     1441 tcggctggaa ggacatcaaa gaatggtacg agatgctgat gggccactgc acctacttcc
     1501 ccgaggaact gcggagcgtg aagtacgcct acaacgccga cctgtacaac gccctgaacg
     1561 acctgaacaa tctcgtgatc accagggacg agaacgagaa gctggaatat tacgagaagt
     1621 tccagatcat cgagaacgtg ttcaagcaga agaagaagcc caccctgaag cagatcgcca
     1681 aagaaatcct cgtgaacgaa gaggatatta agggctacag agtgaccagc accggcaagc
     1741 ccgagttcac caacctgaag gtgtaccacg acatcaagga cattaccgcc cggaaagaga
     1801 ttattgagaa cgccgagctg ctggatcaga ttgccaagat cctgaccatc taccagagca
     1861 gcgaggacat ccaggaagaa ctgaccaatc tgaactccga gctgacccag gaagagatcg
     1921 agcagatctc taatctgaag ggctataccg gcacccacaa cctgagcctg aaggccatca
     1981 acctgatcct ggacgagctg tggcacacca acgacaacca gatcgctatc ttcaaccggc
     2041 tgaagctggt gcccaagaag gtggacctgt cccagcagaa agagatcccc accaccctgg
     2101 tggacgactt catcctgagc cccgtcgtga agagaagctt catccagagc atcaaagtga
     2161 tcaacgccat catcaagaag tacggcctgc ccaacgacat cattatcgag ctggcccgcg
     2221 agaagaactc caaggacgcc cagaaaatga tcaacgagat gcagaagcgg aaccggcaga
     2281 ccaacgagcg gatcgaggaa atcatccgga ccaccggcaa agagaacgcc aagtacctga
     2341 tcgagaagat caagctgcac gacatgcagg aaggcaagtg cctgtacagc ctggaagcca
     2401 tccctctgga agatctgctg aacaacccct tcaactatga ggtggaccac atcatcccca
     2461 gaagcgtgtc cttcgacaac agcttcaaca acaaggtgct cgtgaagcag gaagaagcca
     2521 gcaagaaggg caaccggacc ccattccagt acctgagcag cagcgacagc aagatcagct
     2581 acgaaacctt caagaagcac atcctgaatc tggccaaggg caagggcaga atcagcaaga
     2641 ccaagaaaga gtatctgctg gaagaacggg acatcaacag gttctccgtg cagaaagact
     2701 tcatcaaccg gaacctggtg gataccagat acgccaccag aggcctgatg aacctgctgc
     2761 ggagctactt cagagtgaac aacctggacg tgaaagtgaa gtccatcaat ggcggcttca
     2821 ccagctttct gcggcggaag tggaagttta agaaagagcg gaacaagggg tacaagcacc
     2881 acgccgagga cgccctgatc attgccaacg ccgatttcat cttcaaagag tggaagaaac
     2941 tggacaaggc caaaaaagtg atggaaaacc agatgttcga ggaaaagcag gccgagagca
     3001 tgcccgagat cgaaaccgag caggagtaca aagagatctt catcaccccc caccagatca
     3061 agcacattaa ggacttcaag gactacaagt acagccaccg ggtggacaag aagcctaata
     3121 gagagctgat taacgacacc ctgtactcca cccggaagga cgacaagggc aacaccctga
     3181 tcgtgaacaa tctgaacggc ctgtacgaca aggacaatga caagctgaaa aagctgatca
     3241 acaagagccc cgaaaagctg ctgatgtacc accacgaccc ccagacctac cagaaactga
     3301 agctgattat ggaacagtac ggcgacgaga agaatcccct gtacaagtac tacgaggaaa
     3361 ccgggaacta cctgaccaag tactccaaaa aggacaacgg ccccgtgatc aagaagatta
     3421 agtattacgg caacaaactg aacgcccatc tggacatcac cgacgactac cccaacagca
     3481 gaaacaaggt cgtgaagctg tccctgaagc cctacagatt cgacgtgtac ctggacaatg
     3541 gcgtgtacaa gttcgtgacc gtgaagaatc tggatgtgat caaaaaagaa aactactacg
     3601 aagtgaatag caagtgctat gaggaagcta agaagctgaa gaagatcagc aaccaggccg
     3661 agtttatcgc ctccttctac aacaacgatc tgatcaagat caacggcgag ctgtatagag
     3721 tgatcggcgt gaacaacgac ctgctgaacc ggatcgaagt gaacatgatc gacatcacct
     3781 accgcgagta cctggaaaac atgaacgaca agaggccccc caggatcatt aagacaatcg
     3841 cctccaagac ccagagcatt aagaagtaca gcacagacat tctgggcaac ctgtatgaag
     3901 tgaaatctaa gaagcaccct cagatcatca aaaagggcaa aaggccggcg gccacgaaaa
     3961 aggccggcca ggcaaaaaag aaaaagggag gaggatcttt atctgggaag aaggcggcag
     4021 ccgcggcggc ggcggctgca gcggcagcaa ccgggacgga ggctggccct gggacagcag
     4081 gcggctccga gaacgggtct gaggtggccg cgcagcccgc gggcctgtcg ggcccagccg
     4141 aggtcgggcc gggggcggtg ggggagcgca caccccgcaa gaaagagcct ccgcgggcct
     4201 cgccccccgg gggcctggcg gaaccgccgg ggtccgcagg gcctcaggcc ggccctactg
     4261 tcgtgcctgg gtctgcgacc cccatggaaa ctggaatagc agagactccg gaggggcgtc
     4321 ggaccagccg gcgcaagcgg gcgaaggtag agtacagaga gatggatgaa agcttggcca
     4381 acctctcaga agatgagtat tattcagaag aagagagaaa tgccaaagca gagaaggaaa
     4441 agaagcttcc cccaccaccc cctcaagccc cacctgagga agaaaatgaa agtgagcctg
     4501 aagaaccatc gggtgtggag ggcgcagctt tccagagccg acttcctcat gaccggatga
     4561 cttctcaaga agcagcctgt tttccagata ttatcagtgg accacaacag acccagaagg
     4621 tttttctttt cattagaaac cgcacactgc agttgtggtt ggataatcca aagattcagc
     4681 tgacatttga ggctactctc caacaattag aagcacctta taacagtgat actgtgcttg
     4741 tccaccgagt tcacagttat ttagagcgtc atggtcttat caacttcggc atctataaga
     4801 ggataaaacc cctaccaact aaaaagacag gaaaggtaat tattataggc tctggggtct
     4861 caggcttggc agcagctcga cagttacaaa gttttggaat ggatgtcaca cttttggaag
     4921 ccagggatcg tgtgggtgga cgagttgcca catttcgcaa aggaaactat gtagctgatc
     4981 ttggagccat ggtggtaaca ggtcttggag ggaatcctat ggctgtggtc agcaaacaag
     5041 taaatatgga actggccaag atcaagcaaa aatgcccact ttatgaagcc aacggacaag
     5101 ctgttcctaa agagaaagat gaaatggtag agcaagagtt taaccggttg ctagaagcta
     5161 catcttacct tagtcatcaa ctagacttca atgtcctcaa taataagcat gtgtcccttg
     5221 gccaggcatt ggaagttgtc attcagttac aagagaagca tgtcaaagat gagcagattg
     5281 aacattggaa gaagatagtg aaaactcagg aagaattgaa agaacttctt aataagatgg
     5341 taaatttgaa agagaaaatt aaagaactcc atcagcaata caaagaagca tctgaagtaa
     5401 agccacccag agatattact gccgagttct tagtgaaaag caaacacagg gatctgaccg
     5461 ccctatgcaa ggaatatgat gaattagctg aaacacaagg aaagctagaa gaaaaacttc
     5521 aggagttgga agcgaatccc ccaagtgatg tatatctctc atcaagagac agacaaatac
     5581 ttgattggca ttttgcaaat cttgaatttg ctaatgccac acctctctca actctctccc
     5641 ttaagcactg ggatcaggat gatgactttg agttcactgg cagccacctg acagtaagga
     5701 atggctactc gtgtgtgcct gtggctttag cagaaggcct agacattaaa ctgaatacag
     5761 cagtgcgaca ggttcgctac acggcttcag gatgtgaagt gatagctgtg aatacccgct
     5821 ccacgagtca aacctttatt tataaatgcg acgcagttct ctgtaccctt cccctgggtg
     5881 tgctgaagca gcagccacca gccgttcagt ttgtgccacc tctccctgag tggaaaacat
     5941 ctgcagtcca aaggatggga tttggcaacc ttaacaaggt ggtgttgtgt tttgatcggg
     6001 tgttctggga tccaagtgtc aatttgttcg ggcatgttgg cagtacgact gccagcaggg
     6061 gtgagctctt cctcttctgg aacctctata aagctccaat actgttggca ctagtggcag
     6121 gagaagctgc tggtatcatg gaaaacataa gtgacgatgt gattgttggc cgatgcctgg
     6181 ccattctcaa agggattttt ggtagcagtg cagtacctca gcccaaagaa actgtggtgt
     6241 ctcgttggcg tgctgatccc tgggctcggg gctcttattc ctatgttgct gcaggatcat
     6301 ctggaaatga ctatgattta atggctcagc caatcactcc tggcccctcg attccaggtg
     6361 ccccacagcc gattccacga ctcttctttg cgggagaaca tacgatccgt aactacccag
     6421 ccacagtgca tggtgctctg ctgagtgggc tgcgagaagc gggaagaatt gcagaccagt
     6481 ttttgggggc catgtatacg ctgcctcgcc aggccacacc aggtgttcct gcacagcagt
     6541 ccccaagcat gggcgcaact ttgtataata aagttgaacg agaaacgtaa aatgatataa
     6601 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     6661 aacatatcca gtcactatga atcaactact tagatggtat tagtgacctg tactgcaggg
     6721 cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     6781 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     6841 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     6901 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     6961 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     7021 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     7081 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     7141 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     7201 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     7261 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     7321 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     7381 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     7441 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     7501 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     7561 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     7621 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     7681 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     7741 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     7801 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7981 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     8041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     8101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     8161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     8221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     8281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8461 cctggccttt tgctggcctt ttgctcacat gtt
//
