LOCUS       pCR8_sgCasRxMatu        3172 bp ds-DNA     circular     23-SEP-2018
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          1..675
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          683..859
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          1050..3172
                     /organism="pCR8/GW/TOPO"
                     /mol_type="other DNA"
                     /label=source:pCR8/GW/TOPO(1)
                     /ApEinfo_label=source:pCR8/GW/TOPO
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     terminator      complement(268..295)
                     /label=rrnB_T2_terminator
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    933..962
                     /label=CasRx-DR
                     /ApEinfo_fwdcolor=cyan
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     terminator      complement(427..470)
                     /label=rrnB_T1_terminator
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    522..544
                     /label=M13_pUC_fwd_primer
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    537..553
                     /label=M13_forward20_primer
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    complement(569..668)
                     /label=attL1
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    671..675
                     /label=EcoRIHS
                     /ApEinfo_fwdcolor=#ff8b8e
                     /ApEinfo_revcolor=#ff8b8e
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    1051..1150
                     /label=attL2
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        complement(1168..1186)
                     /label=T7_promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        683..859
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /label=U6 promoter
                     /ApEinfo_fwdcolor=#346ee0
                     /ApEinfo_revcolor=#346ee0
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        860..923
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /label=U6 promoter(1)
                     /ApEinfo_label=U6 promoter
                     /ApEinfo_fwdcolor=#346ee0
                     /ApEinfo_revcolor=#346ee0
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    complement(1187..1205)
                     /label=M13_reverse_primer
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             1419..2429
                     /gene="spect"
                     /note="ORF frame 2"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQA
                     RYAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLL
                     AVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVE
                     VTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVAL
                     VGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIA
                     PKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVV
                     GK*"
                     /label=spect
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     primer_bind     683..703
                     /note="Human U6 promoter, forward primer"
                     /label=hU6-F
                     /ApEinfo_fwdcolor=#14c0bd
                     /ApEinfo_revcolor=#4ec02b
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     gene            1419..2429
                     /gene="spect"
                     /label=spect(1)
                     /ApEinfo_label=spect
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      2506..3125
                     /label=pBR322_origin
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     primer_bind     854..859
                     /note="Human U6 promoter, forward primer"
                     /label=LKO.1 5'
                     /ApEinfo_fwdcolor=#14c0bd
                     /ApEinfo_revcolor=#4ec02b
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          860..931
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct(1)
                     /ApEinfo_label=source:synthetic DNA construct
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     source          1021..1027
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct(2)
                     /ApEinfo_label=source:synthetic DNA construct
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     primer_bind     860..873
                     /note="Human U6 promoter, forward primer"
                     /label=LKO.1 5'(1)
                     /ApEinfo_label=LKO.1 5'
                     /ApEinfo_fwdcolor=#14c0bd
                     /ApEinfo_revcolor=#4ec02b
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
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 ACAACGaTCA AATCCGCTCC CGGCGGATTT GTCCTACTCA GGAGAGCGTT CACCGACAAA
      421 CAACAGATAA AACGAaaggc ccagtattcc 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 AGCAGGCTCC GAATTcACCG GTgagggcct atttcccatg attccttcat atttgcatat
      721 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt
      781 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
      841 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
      901 tttatatatc ttgtggaaag gacgaaacac cgAACCCCTA CCAACTGGTC GGGGTTTGAA
      961 ACGAGAATTC TAGTAGGGAT GTAGCAAGTA AACCCCTACC AACTGGTCGG GGTTTGAAAC
     1021 tttttttGGC GCGCCTTAAT TAAGAATTCG ACCCAGCTTT CTTGTACAAA GTTGGCATTA
     1081 TAAAAAATAA TTGCTCATCA ATTTGTTGCA ACGAACAGGT CACTATCAGT CAAAATAAAA
     1141 TCATTATTTG CCATCCAGCT GATATCCCCT ATAGTGAGTC GTATTACATG GTCATAGCTG
     1201 TTTCCTGGCA GCTCTGGCCC GTGTCTCAAA ATCTCTGATG TTACATTGCA CAAGATAAAA
     1261 ATATATCATC ATGCCTCCTC TAGACCAGCC AGGACAGAAA TGCCTCGACT TCGCTGCTGC
     1321 CCAAGGTTGC CGGGTGACGC ACACCGTGGA AACGGATGAA GGCACGAACC CAGTGGACAT
     1381 AAGCCTGTTC GGTTCGTAAG CTGTAATGCA AGTAGCGTAT GCGCTCACGC AACTGGTCCA
     1441 GAACCTTGAC CGAACGCAGC GGTGGTAACG GCGCAGTGGC GGTTTTCATG GCTTGTTATG
     1501 ACTGTTTTTT TGGGGTACAG TCTATGCCTC GGGCATCCAA GCAGCAAGCG CGTTACGCCG
     1561 TGGGTCGATG TTTGATGTTA TGGAGCAGCA ACGATGTTAC GCAGCAGGGC AGTCGCCCTA
     1621 AAACAAAGTT AAACATCATG AGGGAAGCGG TGATCGCCGA AGTATCGACT CAACTATCAG
     1681 AGGTAGTTGG CGTCATCGAG CGCCATCTCG AACCGACGTT GCTGGCCGTA CATTTGTACG
     1741 GCTCCGCAGT GGATGGCGGC CTGAAGCCAC ACAGTGATAT TGATTTGCTG GTTACGGTGA
     1801 CCGTAAGGCT TGATGAAACA ACGCGGCGAG CTTTGATCAA CGACCTTTTG GAAACTTCGG
     1861 CTTCCCCTGG AGAGAGCGAG ATTCTCCGCG CTGTAGAAGT CACCATTGTT GTGCACGACG
     1921 ACATCATTCC GTGGCGTTAT CCAGCTAAGC GCGAACTGCA ATTTGGAGAA TGGCAGCGCA
     1981 ATGACATTCT TGCAGGTATC TTCGAGCCAG CCACGATCGA CATTGATCTG GCTATCTTGC
     2041 TGACAAAAGC AAGAGAACAT AGCGTTGCCT TGGTAGGTCC AGCGGCGGAG GAACTCTTTG
     2101 ATCCGGTTCC TGAACAGGAT CTATTTGAGG CGCTAAATGA AACCTTAACG CTATGGAACT
     2161 CGCCGCCCGA CTGGGCTGGC GATGAGCGAA ATGTAGTGCT TACGTTGTCC CGCATTTGGT
     2221 ACAGCGCAGT AACCGGCAAA ATCGCGCCGA AGGATGTCGC TGCCGACTGG GCAATGGAGC
     2281 GCCTGCCGGC CCAGTATCAG CCCGTCATAC TTGAAGCTAG ACAGGCTTAT CTTGGACAAG
     2341 AAGAAGATCG CTTGGCCTCG CGCGCAGATC AGTTGGAAGA ATTTGTCCAC TACGTGAAAG
     2401 GCGAGATCAC CAAGGTAGTC GGCAAATAAC CCTCGAGCCA CCCATGACCA AAATCCCTTA
     2461 ACGTGAGTTA CGCGTCGTTC CACTGAGCGT CAGACCCCGT AGAAAAGATC AAAGGATCTT
     2521 CTTGAGATCC TTTTTTTCTG CGCGTAATCT GCTGCTTGCA AACAAAAAAA CCACCGCTAC
     2581 CAGCGGTGGT TTGTTTGCCG GATCAAGAGC TACCAACTCT TTTTCCGAAG GTAACTGGCT
     2641 TCAGCAGAGC GCAGATACCA AATACTGTCC TTCTAGTGTA GCCGTAGTTA GGCCACCACT
     2701 TCAAGAACTC TGTAGCACCG CCTACATACC TCGCTCTGCT AATCCTGTTA CCAGTGGCTG
     2761 CTGCCAGTGG CGATAAGTCG TGTCTTACCG GGTTGGACTC AAGACGATAG TTACCGGATA
     2821 AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT CGTGCACACA GCCCAGCTTG GAGCGAACGA
     2881 CCTACACCGA ACTGAGATAC CTACAGCGTG AGCATTGAGA AAGCGCCACG CTTCCCGAAG
     2941 GGAGAAAGGC GGACAGGTAT CCGGTAAGCG GCAGGGTCGG AACAGGAGAG CGCACGAGGG
     3001 AGCTTCCAGG GGGAAACGCC TGGTATCTTT ATAGTCCTGT CGGGTTTCGC CACCTCTGAC
     3061 TTGAGCGTCG ATTTTTGTGA TGCTCGTCAG GGGGGCGGAG CCTATGGAAA AACGCCAGCA
     3121 ACGCGGCCTT TTTACGGTTC CTGGCCTTTT GCTGGCCTTT TGCTCACATG TT
//
