LOCUS       sequence-237395-        3264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMAGIC L3-L2 entry plasmid, contains 3xHA tag+polyA; empty 
            hU6-driven SaCas9 gRNA scaffold for 3-/4-component MultiSite Gateway
            Pro assembly. Allows C-term epitope fusion to dCas9 w/ gRNA 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3264)
  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 3264)
  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: JI501: pMAGIC (L3-L2) 3x HA epitope tag + poly...
FEATURES             Location/Qualifiers
     source          1..3264
                     /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    570..665
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             677..757
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(785..802)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    791..998
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1005..1253
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1005..1025
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1176..1195
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(1376..1475)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1505..1524)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1506..1524)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1529..1545)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1529..1545)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1647..1666)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1658..2467
                     /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(1725..1744)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2614..3202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3103..3122
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa
      661 agttggtggc ggatcctacc catacgatgt tccagattac gcttacccat acgatgttcc
      721 agattacgct tacccatacg atgttccaga ttacgcttaa gaattcctag agctcgctga
      781 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
      841 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
      901 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
      961 ggggaggatt gggaagagaa tagcaggcat gctggggagg taccgagggc ctatttccca
     1021 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     1081 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     1141 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     1201 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accggagacc
     1261 acggcaggtc tcagtttaag tactctgtgc tggaaacagc acagaatcta ctaaacaagg
     1321 caaaatgccg tgtttatctc gtcaacttgt tggcgagatt tttgcggccg caggaaccca
     1381 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     1441 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagg gcggccgcga
     1501 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1561 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1621 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1681 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1741 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1801 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1861 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1921 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1981 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2041 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2101 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2161 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2221 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2281 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2341 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2401 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2461 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2521 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2581 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2641 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2701 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2761 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2821 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2881 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2941 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3001 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3061 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3241 ttgctggcct tttgctcaca tgtt
//
