LOCUS       Exported                7791 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pMAGIC R4-R3 entry plasmid, contains 2x NLS Sa-dCas9 fused to the 
            p300 catalytic core for 3 or 4-component MultiSite Gateway Pro 
            assembly..
ACCESSION   .
VERSION     .
KEYWORDS    IF405: pMAGIC (R4-R3) NLS-Sa dCas9-NLS-p300core
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7791)
  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 7791)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7791
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      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..727)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      726..735
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     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"
     CDS             3999..5849
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      5613..5622
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    5854..5977
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6032..6051)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6033..6051)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6056..6072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6056..6072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6174..6193)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6185..6994
                     /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(6252..6271)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7141..7729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7630..7649
                     /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 gcgggagcat tttcaaacca gaagaactac
     4021 gacaggcact gatgccaact ttggaggcac tttaccgtca ggatccagaa tcccttccct
     4081 ttcgtcaacc tgtggaccct cagcttttag gaatccctga ttactttgat attgtgaaga
     4141 gccccatgga tctttctacc attaagagga agttagacac tggacagtat caggagccct
     4201 ggcagtatgt cgatgatatt tggcttatgt tcaataatgc ctggttatat aaccggaaaa
     4261 catcacgggt atacaaatac tgctccaagc tctctgaggt ctttgaacaa gaaattgacc
     4321 cagtgatgca aagccttgga tactgttgtg gcagaaagtt ggagttctct ccacagacac
     4381 tgtgttgcta cggcaaacag ttgtgcacaa tacctcgtga tgccacttat tacagttacc
     4441 agaacaggta tcatttctgt gagaagtgtt tcaatgagat ccaaggggag agcgtttctt
     4501 tgggggatga cccttcccag cctcaaacta caataaataa agaacaattt tccaagagaa
     4561 aaaatgacac actggatcct gaactgtttg ttgaatgtac agagtgcgga agaaagatgc
     4621 atcagatctg tgtccttcac catgagatca tctggcctgc tggattcgtc tgtgatggct
     4681 gtttaaagaa aagtgcacga actaggaaag aaaataagtt ttctgctaaa aggttgccat
     4741 ctaccagact tggcaccttt ctagagaatc gtgtgaatga ctttctgagg cgacagaatc
     4801 accctgagtc aggagaggtc actgttagag tagttcatgc ttctgacaaa accgtggaag
     4861 taaaaccagg catgaaagca aggtttgtgg acagtggaga gatggcagaa tcctttccat
     4921 accgaaccaa agccctcttt gcctttgaag aaattgatgg tgttgacctg tgcttctttg
     4981 gcatgcatgt tcaagagtat ggctctgact gccctccacc caaccagagg agagtataca
     5041 tatcttacct cgatagtgtt catttcttcc gtcctaaatg cttgaggact gcagtctatc
     5101 atgaaatcct aattggatat ttagaatatg tcaagaaatt aggttacaca acagggcata
     5161 tttgggcatg tccaccaagt gagggagatg attatatctt ccattgccat cctcctgacc
     5221 agaagatacc caagcccaag cgactgcagg aatggtacaa aaaaatgctt gacaaggctg
     5281 tatcagagcg tattgtccat gactacaagg atatttttaa acaagctact gaagatagat
     5341 taacaagtgc aaaggaattg ccttatttcg agggtgattt ctggcccaat gttctggaag
     5401 aaagcattaa ggaactggaa caggaggaag aagagagaaa acgagaggaa aacaccagca
     5461 atgaaagcac agatgtgacc aagggagaca gcaaaaatgc taaaaagaag aataataaga
     5521 aaaccagcaa aaataagagc agcctgagta ggggcaacaa gaagaaaccc gggatgccca
     5581 atgtatctaa cgacctctca cagaaactat atgccaccat ggagaagcat aaagaggtct
     5641 tctttgtgat ccgcctcatt gctggccctg ctgccaactc cctgcctccc attgttgatc
     5701 ctgatcctct catcccctgc gatctgatgg atggtcggga tgcgtttctc acgctggcaa
     5761 gggacaagca cctggagttc tcttcactcc gaagagccca gtggtccacc atgtgcatgc
     5821 tggtggagct gcacacgcag agccaggacg gcgcaacttt gtataataaa gttgaacgag
     5881 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     5941 cataatactg taaaacacaa catatccagt cactatgaat caactactta gatggtatta
     6001 gtgacctgta ctgcagggcg gccgcgatat cccctatagt gagtcgtatt acatggtcat
     6061 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     6121 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     6181 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     6241 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     6301 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     6361 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     6421 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     6481 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     6541 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     6601 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     6661 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     6721 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     6781 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     6841 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     6901 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     6961 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     7021 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     7081 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7141 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7201 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7261 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7321 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7381 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7441 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7501 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7561 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7621 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7681 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7741 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
