LOCUS       sequence-314395-        3266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMAGIC L3-L2 entry plasmid, contains 3xHA tag+polyA; hU6-driven 
            SaCas9 TRE#2 gRNA for 3-/4-component MultiSite Gateway Pro assembly.
            Allows C-term HA epitope fusion to dCas9 w/ TRE#2 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: LB101: pMAGIC (L3-L2) 3x HA epitope tag + poly...
FEATURES             Location/Qualifiers
     source          1..3266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(68..154)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     203..225
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     217..234
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     218..234
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    251..346
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             358..438
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(466..483)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    472..679
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        686..934
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     686..706
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     857..876
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(1059..1158)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1188..1207)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1189..1207)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1212..1228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1212..1228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1330..1349)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1341..2150
                     /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(1408..1427)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2297..2885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2786..2805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3039..3056
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3215..3242)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
       61 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
      121 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
      181 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
      241 ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt
      301 gatgagcaat gcttttttat aatgccaact ttgtataata aagttggtgg cggatcctac
      361 ccatacgatg ttccagatta cgcttaccca tacgatgttc cagattacgc ttacccatac
      421 gatgttccag attacgctta agaattccta gagctcgctg atcagcctcg actgtgcctt
      481 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
      541 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
      601 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga
      661 atagcaggca tgctggggag gtaccgaggg cctatttccc atgattcctt catatttgca
      721 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
      781 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
      841 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
      901 ggctttatat atcttgtgga aaggacgaaa caccgcatcg ttctctatca ctgatagttt
      961 aagtactctg tgctggaaac agcacagaat ctactaaaca aggcaaaatg ccgtgtttat
     1021 ctcgtcaact tgttggcgag atttttgcgg ccgcaggaac ccagctttct tgtacaaagt
     1081 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1141 aaataaaatc attatttgcc atccagctgc agggcggccg cgatatcccc tatagtgagt
     1201 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     1261 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1321 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     1381 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     1441 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     1501 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     1561 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     1621 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     1681 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     1741 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     1801 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     1861 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     1921 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     1981 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     2041 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     2101 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     2161 ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat
     2221 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     2281 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2341 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2401 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2461 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2521 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2581 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2641 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2701 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2761 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2821 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2881 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2941 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3001 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3061 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3121 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     3181 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     3241 ctgcttagtt tgatgcctgg cagttt
//
