LOCUS       sequence-237392-        5367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMAGIC R4-R3 entry plasmid, contains 2x NLS/LSD1 scaffold enabling 
            addition of dCas9 mutants into pMAGIC. dCas9 can be inserted via 
            unique SrfI/PmeI restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5367)
  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 5367)
  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: KL202: pMAGIC (R4-R3) NLS-(SrfI/PmeI)-NLS-LSD1
FEATURES             Location/Qualifiers
     source          1..5367
                     /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             813..860
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    3430..3553
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3608..3627)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3609..3627)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3632..3648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3632..3648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3750..3769)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3761..4570
                     /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(3828..3847)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4717..5305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5206..5225
                     /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 cccgggcgag accatgctcg cggtctcagt ttaaacggcc ggcagccacg aagaaggccg
      841 gccaggcaaa aaagaaaaag ggaggaggat ctttatctgg gaagaaggcg gcagccgcgg
      901 cggcggcggc tgcagcggca gcaaccggga cggaggctgg ccctgggaca gcaggcggct
      961 ccgagaacgg gtctgaggtg gccgcgcagc ccgcgggcct gtcgggccca gccgaggtcg
     1021 ggccgggggc ggtgggggag cgcacacccc gcaagaaaga gcctccgcgg gcctcgcccc
     1081 ccgggggcct ggcggaaccg ccggggtccg cagggcctca ggccggccct actgtcgtgc
     1141 ctgggtctgc gacccccatg gaaactggaa tagcagagac tccggagggg cgtcggacca
     1201 gccggcgcaa gcgggcgaag gtagagtaca gagagatgga tgaaagcttg gccaacctct
     1261 cagaagatga gtattattca gaagaagaga gaaatgccaa agcagagaag gaaaagaagc
     1321 ttcccccacc accccctcaa gccccacctg aggaagaaaa tgaaagtgag cctgaagaac
     1381 catcgggtgt ggagggcgca gctttccaga gccgacttcc tcatgaccgg atgacttctc
     1441 aagaagcagc ctgttttcca gatattatca gtggaccaca acagacccag aaggtttttc
     1501 ttttcattag aaaccgcaca ctgcagttgt ggttggataa tccaaagatt cagctgacat
     1561 ttgaggctac tctccaacaa ttagaagcac cttataacag tgatactgtg cttgtccacc
     1621 gagttcacag ttatttagag cgtcatggtc ttatcaactt cggcatctat aagaggataa
     1681 aacccctacc aactaaaaag acaggaaagg taattattat aggctctggg gtctcaggct
     1741 tggcagcagc tcgacagtta caaagttttg gaatggatgt cacacttttg gaagccaggg
     1801 atcgtgtggg tggacgagtt gccacatttc gcaaaggaaa ctatgtagct gatcttggag
     1861 ccatggtggt aacaggtctt ggagggaatc ctatggctgt ggtcagcaaa caagtaaata
     1921 tggaactggc caagatcaag caaaaatgcc cactttatga agccaacgga caagctgttc
     1981 ctaaagagaa agatgaaatg gtagagcaag agtttaaccg gttgctagaa gctacatctt
     2041 accttagtca tcaactagac ttcaatgtcc tcaataataa gcatgtgtcc cttggccagg
     2101 cattggaagt tgtcattcag ttacaagaga agcatgtcaa agatgagcag attgaacatt
     2161 ggaagaagat agtgaaaact caggaagaat tgaaagaact tcttaataag atggtaaatt
     2221 tgaaagagaa aattaaagaa ctccatcagc aatacaaaga agcatctgaa gtaaagccac
     2281 ccagagatat tactgccgag ttcttagtga aaagcaaaca cagggatctg accgccctat
     2341 gcaaggaata tgatgaatta gctgaaacac aaggaaagct agaagaaaaa cttcaggagt
     2401 tggaagcgaa tcccccaagt gatgtatatc tctcatcaag agacagacaa atacttgatt
     2461 ggcattttgc aaatcttgaa tttgctaatg ccacacctct ctcaactctc tcccttaagc
     2521 actgggatca ggatgatgac tttgagttca ctggcagcca cctgacagta aggaatggct
     2581 actcgtgtgt gcctgtggct ttagcagaag gcctagacat taaactgaat acagcagtgc
     2641 gacaggttcg ctacacggct tcaggatgtg aagtgatagc tgtgaatacc cgctccacga
     2701 gtcaaacctt tatttataaa tgcgacgcag ttctctgtac ccttcccctg ggtgtgctga
     2761 agcagcagcc accagccgtt cagtttgtgc cacctctccc tgagtggaaa acatctgcag
     2821 tccaaaggat gggatttggc aaccttaaca aggtggtgtt gtgttttgat cgggtgttct
     2881 gggatccaag tgtcaatttg ttcgggcatg ttggcagtac gactgccagc aggggtgagc
     2941 tcttcctctt ctggaacctc tataaagctc caatactgtt ggcactagtg gcaggagaag
     3001 ctgctggtat catggaaaac ataagtgacg atgtgattgt tggccgatgc ctggccattc
     3061 tcaaagggat ttttggtagc agtgcagtac ctcagcccaa agaaactgtg gtgtctcgtt
     3121 ggcgtgctga tccctgggct cggggctctt attcctatgt tgctgcagga tcatctggaa
     3181 atgactatga tttaatggct cagccaatca ctcctggccc ctcgattcca ggtgccccac
     3241 agccgattcc acgactcttc tttgcgggag aacatacgat ccgtaactac ccagccacag
     3301 tgcatggtgc tctgctgagt gggctgcgag aagcgggaag aattgcagac cagtttttgg
     3361 gggccatgta tacgctgcct cgccaggcca caccaggtgt tcctgcacag cagtccccaa
     3421 gcatgggcgc aactttgtat aataaagttg aacgagaaac gtaaaatgat ataaatatca
     3481 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     3541 tccagtcact atgaatcaac tacttagatg gtattagtga cctgtactgc agggcggccg
     3601 cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     3661 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     3721 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     3781 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3841 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3901 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3961 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     4021 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     4081 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     4141 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     4201 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     4261 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     4321 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     4381 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     4441 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     4501 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     4561 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     4621 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     4681 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4741 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4801 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4861 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4921 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4981 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5041 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5101 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5161 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5221 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5281 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5341 cttttgctgg ccttttgctc acatgtt
//
