LOCUS       Exported                3444 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pMAGIC L1-R5 entry plasmid, contains empty human U6-driven SaCas9 
            gRNA scaffold and RIP for 4-component MultiSite Gateway Pro 
            assembly. Protospacer motif can be inserted after BsaI digestion..
ACCESSION   .
VERSION     .
KEYWORDS    JI601: pMAGIC (L1-R5) hU6::SaCas9 gRNA scaffold; RIP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3444)
  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 3444)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3444
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        679..919
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     679..699
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     850..869
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    1507..1630
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1685..1704)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1686..1704)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1709..1725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1709..1725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1827..1846)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1838..2647
                     /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(1905..1924)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2794..3382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3283..3302
                     /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 tgtacaaaaa
      661 agcaggctta tggtacctga gggcctattt cccatgattc cttcatattt gcatatacga
      721 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
      781 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
      841 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
      901 tatatcttgt ggaaaggacg aaacaccgga gaccacggca ggtctcagtt taagtactct
      961 gtgctggaaa cagcacagaa tctactaaac aaggcaaaat gccgtgttta tctcgtcaac
     1021 ttgttggcga gatttttgcg gccgctgtac atacgactct agaaagactg agctaagaat
     1081 ccagctatca atagaaacta tgaaacagtt ccagggacaa agataccagg tccccaacaa
     1141 ctgcaacttt ctgggaaatg aggtggaaaa tgctcagcca aggaaaaaga gggccttacc
     1201 ctctctggga caatgattgt gctgtgaact gcttcatcag gccatctggc cccttgttaa
     1261 taatctaatt accctaggtc taagtagagt tgttgacgtc caatgagcgc tttctgcaga
     1321 cttagcacta ggcaagtgtt tggaaattac agcttcagcc cctctcgcca tctgcctacc
     1381 tacccctcct agagccctta atgggccaaa cggcaaagtc cagggggcag agaggaggtg
     1441 ctttggacta taaagctagt ggggacccag taactcccaa ccctaagtga ccagctacga
     1501 attccacaac tttgtataca aaagttgaac gagaaacgta aaatgatata aatatcaata
     1561 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
     1621 agtcactatg aatcaactac ttagatggta ttagtgacct gtactgcagg gcggccgcga
     1681 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1741 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1801 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1861 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1921 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1981 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2041 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2101 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2161 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2221 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2281 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2341 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2401 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2461 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2521 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2581 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2641 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2701 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2941 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3421 ttgctggcct tttgctcaca tgtt
//
