LOCUS       sequence-314449-        3266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMAGIC L3-L2 entry plasmid, contains 3xHA tag+polyA; hU6-driven 
            Sp/xCas9 TRE#1 gRNA for 3-/4-component MultiSite Gateway Pro 
            assembly. Allows C-term HA epitope fusion to dCas9 w/ TRE#1 gRNA 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3266)
  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 3266)
  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: LA901: pMAGIC (L3-L2) 3x HA epitope tag + poly...
FEATURES             Location/Qualifiers
     source          1..3266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             98..178
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(206..223)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    212..419
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        426..674
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     426..446
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     597..616
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(799..898)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(928..947)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(929..947)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(952..968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(952..968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1070..1089)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1081..1890
                     /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(1148..1167)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2037..2625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2526..2545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2779..2796
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2955..2982)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3074..3160)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3209..3231
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3223..3240
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3224..3240
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(3257..3266,1..86)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat
       61 aatgccaact ttgtataata aagttggtgg cggatcctac ccatacgatg ttccagatta
      121 cgcttaccca tacgatgttc cagattacgc ttacccatac gatgttccag attacgctta
      181 agaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
      241 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
      301 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
      361 ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag
      421 gtaccgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
      481 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      541 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      601 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      661 aaggacgaaa caccgtgata gagaacgtat gtcggtttca gagctatgct ggaaacagca
      721 tagcaagttg aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
      781 ttttttgcgg ccgcaggaac ccagctttct tgtacaaagt tggcattata agaaagcatt
      841 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
      901 atccagctgc agggcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct
      961 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     1021 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     1081 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     1141 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     1201 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     1261 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     1321 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     1381 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     1441 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     1501 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     1561 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     1621 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     1681 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     1741 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     1801 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     1861 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     1921 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     1981 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2041 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2101 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2161 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2221 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2281 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2341 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2401 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2461 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2521 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2581 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2641 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2701 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2761 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2821 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2881 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     2941 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     3001 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     3061 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     3121 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     3181 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     3241 cttaagctcg ggccccaaat aatgat
//
