LOCUS       sequence-314448-        3266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMAGIC L3-L2 entry plasmid, contains 3xHA tag+polyA; hU6-driven 
            SaCas9 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: JV201: pMAGIC (L3-L2) 3x HA epitope tag + poly...
FEATURES             Location/Qualifiers
     source          1..3266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(44..130)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     193..210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     194..210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    227..322
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             334..414
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(442..459)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    448..655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        662..910
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     662..682
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     833..852
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(1035..1134)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1164..1183)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1165..1183)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1188..1204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1188..1204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1306..1325)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1317..2126
                     /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(1384..1403)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2273..2861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2762..2781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3015..3032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3191..3218)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
       61 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
      121 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
      181 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta
      241 ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg
      301 ccaactttgt ataataaagt tggtggcgga tcctacccat acgatgttcc agattacgct
      361 tacccatacg atgttccaga ttacgcttac ccatacgatg ttccagatta cgcttaagaa
      421 ttcctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
      481 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
      541 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
      601 tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct ggggaggtac
      661 cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      721 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      781 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      841 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      901 acgaaacacc gatcagtgat agagaacgta tggtttaagt actctgtgct ggaaacagca
      961 cagaatctac taaacaaggc aaaatgccgt gtttatctcg tcaacttgtt ggcgagattt
     1021 ttgcggccgc aggaacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1081 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     1141 agctgcaggg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     1201 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     1261 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     1321 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     1381 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     1441 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     1501 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     1561 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     1621 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     1681 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1741 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1801 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1861 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     1921 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     1981 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     2041 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     2101 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     2161 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     2221 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2281 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2341 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2401 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2461 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2521 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2581 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2641 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2701 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2761 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2821 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2881 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2941 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3001 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     3061 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     3121 ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg
     3181 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt
     3241 ttatggcggg cgtcctgccc gccacc
//
