LOCUS       pCFD4d                  4575 bp    DNA     circular SYN 18-Apr-2014
DEFINITION  pCFD4-U6:1_U6:3tandemgRNAs
ACCESSION   -
KEYWORDS    -
SOURCE      -
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          1337..1366
                     /label=source:pCFD4-U6:1_U6:3tandemgRNAs
                     /mol_type=other DNA
                     /note="/ApEinfo_fwdcolor=#ffffff;
                     /ApEinfo_revcolor=#ffffff;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /organism="pCFD4-U6:1_U6:3tandemgRNAs"
     primer_bind     1..28
                     /label=U6-1_forward primer
                     /note="/ApEinfo_fwdcolor=#808000;
                     /ApEinfo_revcolor=#808000;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        8..803
                     /label=U6.1 promoter
                     /note="/ApEinfo_fwdcolor=#6699ff;
                     /ApEinfo_revcolor=#6699ff;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     804..821
                     /note="BbsI site for gRNA insertion"
     misc_feature    806..811
                     /label=BbsI
                     /note="/ApEinfo_fwdcolor=#4c4c4c;
                     /ApEinfo_revcolor=#4c4c4c;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    814..819
                     /label=BbsI(1)
                     /note="/ApEinfo_label=BbsI; /ApEinfo_fwdcolor=#4c4c4c;
                     /ApEinfo_revcolor=#4c4c4c;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    822..897
                     /note="sgRNA scaffold"
     misc_RNA        822..897
                     /label=gRNA_core
                     /note="/ApEinfo_fwdcolor=#ff0000;
                     /ApEinfo_revcolor=#ff0000;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        904..1318
                     /label=U6.3 promoter
                     /note="/ApEinfo_fwdcolor=#6699ff;
                     /ApEinfo_revcolor=#6699ff;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(1166..1184)
                     /label=U6-1_reverse primer
                     /note="/ApEinfo_fwdcolor=#808000;
                     /ApEinfo_revcolor=#808000;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     1208..1226
                     /label=U6-3_forward primer
                     /note="/ApEinfo_fwdcolor=#808000;
                     /ApEinfo_revcolor=#808000;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     1319..1336
                     /note="BbsI site for gRNA insertion"
     misc_feature    1321..1326
                     /label=BbsI(2)
                     /note="/ApEinfo_label=BbsI; /ApEinfo_fwdcolor=#4c4c4c;
                     /ApEinfo_revcolor=#4c4c4c;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1329..1334
                     /label=BbsI(3)
                     /note="/ApEinfo_label=BbsI; /ApEinfo_fwdcolor=#4c4c4c;
                     /ApEinfo_revcolor=#4c4c4c;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1337..1412
                     /note="sgRNA scaffold"
     misc_RNA        1337..1412
                     /label=gRNA_core(1)
                     /note="/ApEinfo_label=gRNA_core;
                     /ApEinfo_fwdcolor=#ff0000; /ApEinfo_revcolor=#ff0000;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(1629..1648)
                     /label=U6-3_reverse primer
                     /note="/ApEinfo_fwdcolor=#808000;
                     /ApEinfo_revcolor=#808000;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(1721..1740)
                     /label=T3
                     /note="/ApEinfo_fwdcolor=cyan; /ApEinfo_revcolor=green;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(1758..1778)
                     /label=M13-rev
                     /note="/ApEinfo_fwdcolor=cyan; /ApEinfo_revcolor=green;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_binding    complement(1762..1779)
                     /note="M13_R"
     misc_binding    complement(1784..1806)
                     /label=LacO
                     /note="/ApEinfo_fwdcolor=#6495ed;
                     /ApEinfo_revcolor=#6495ed;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     operon          complement(1784..1806)
                     /note="LacO"
     oriT            complement(2105..2778)
                     /note="pUC origin"
     gene            complement(2922..3782)
                     /gene="bla"
     CDS             complement(2923..3783)
                     /label=AmpR
                     /note="/ApEinfo_fwdcolor=#ffcc66;
                     /ApEinfo_revcolor=#ffcc66;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     gene            complement(2923..3783)
                     /gene="bla"
     promoter        complement(3824..3852)
                     /note="AmpR_promoter"
     misc_feature    complement(3914..4369)
                     /note="F1 intergenic region"
     rep_origin      complement(3914..4369)
                     /label=F1 ori
                     /note="/ApEinfo_fwdcolor=gray50; /ApEinfo_revcolor=gray50;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     4510..4527
                     /label=M13-fwd
                     /note="/ApEinfo_fwdcolor=cyan; /ApEinfo_revcolor=green;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_binding    4511..4527
                     /note="M13_F"
     primer_bind     4526..4553
                     /label=T7
                     /note="/ApEinfo_fwdcolor=cyan; /ApEinfo_revcolor=green;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        4534..4553
                     /note="T7 promoter"
     misc_feature    4538
                     /note="T7 transcription initiation site"
     misc_feature    4566..4575
                     /label=(BglII-HindIIIjunct_plsConf)
                     /note="/ApEinfo_fwdcolor=#4c4c4c;
                     /ApEinfo_revcolor=#4c4c4c;
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
BASE COUNT      1216 A      1083 C      1054 G      1222 T         0 OTHER
ORIGIN      ?
        1 GAATTCATTT TCAACGTCCT CGATAGTATA GTGGTTAGTA TCCCCGCCTG TCACGCGGGA
       61 GACCGGGGTT CAATTCCCCG TCGGGGAGAA TCTGTGATTC TTTTTTTTTT TCTTTTACTT
      121 TGTTATATAA ACAATTTTTG TTTTAATTGA ATCTAATTTG CCATTGCTTT TAGGAATCTC
      181 AGGCATCCAG CAAGCGTTTG TCCGCCGAAT CGCCCATCAG TGAAGAAGAT CCTGTGGCGG
      241 CTACGAAAAT CTCCCCGGCC ATGTCGGCCT CCACCTCCAG CGAAAAACCC ATCAGCGAGC
      301 TGGCCACCTC TGTGCTGACC CACCGCTTTC CAGACTCCAC CTCCTCACCC GGCGAACATG
      361 GCCTTGGACG AATGCAGTTG TCGATCCGCT ACAGCGCCCA GCGTCAAAAA CTAGACGTGA
      421 CCATACACAA AATCCAGAAG ATACCACTTC GCGATCCCAG CAATATCCCC GATCCGTATG
      481 TTAAGCTGTA TCTGTTGCCT GGACGCACCA AGGAGTCGAA ACGCAAGACG AGCGTGATCA
      541 AGGACAACTG CAACCCCGTC TACGATGCAT CCTTTGAGTA CCTGATTTCC ATTGCCGAAC
      601 TCAGGCAGAC GGAACTGGAG GTGACGGTGT GCACCCAAAA GGGATTCCTA TCCGGCGGTA
      661 GTCCCATCAT TGGCATGGTA GGTACCCGAA AGCAACCCCT TAGTTACAGA CACAGCGCGT
      721 ACGTCCTTCG CATCCTTATG ATTCCCAAGT ACATATTCTG CAAGAGTACA GTATATATAG
      781 GAAAGATATC CGGGTGAACT TCGGGGTCTT CGAGAAGACC TGTTTTAGAG CTAGAAATAG
      841 CAAGTTAAAA TAAGGCTAGT CCGTTATCAA CTTGAAAAAG TGGCACCGAG TCGGTGCTTT
      901 TTTGCTCACC TGTGATTGCT CCTACTCAAA TACAAAAACA TCAAATTTTC TGTCAATAAA
      961 GCATATTTAT TTATATTTAT TTTACAGGAA AGAATTCCTT TTAAAGTGTA TTTTAACCTA
     1021 TAATGAAAAA CGATTAAAAA AAATACATAA AATAATTCGA AAATTTTTGA ATAGCCCAGG
     1081 TTGATAAAAA TTCATTTCAT ACGTTTTATA ACTTATGCCC CTAAGTATTT TTTGACCATA
     1141 GTGTTTCAAT TCTACATTAA TTTTACAGAG TAGAATGAAA CGCCACCTAC TCAGCCAAGA
     1201 GGCGAAAAGG TTAGCTCGCC AAGCAGAGAG GGCGCCAGTG CTCACTACTT TTTATAATTC
     1261 TCAACTTCTT TTTCCAGACT CAGTTCGTAT ATATAGACCT ATTTTCAATT TAACGTCGGG
     1321 GTCTTCGAGA AGACCTGTTT TAGAGCTAGA AATAGCAAGT TAAAATAAGG CTAGTCCGTT
     1381 ATCAACTTGA AAAAGTGGCA CCGAGTCGGT GCTTTTTTGC CTACCTGGAG CCTGAGAGTT
     1441 GTTCAATAAA ATAAAAATGT TTCGTTTTTT TGCTTTCGCC AGTATTTATT ATTTTTCATC
     1501 AATATGTATT CAATTTGGTA TGTATTTAGT AATTGTAATA TATAGACAAT GGTTTTCCGT
     1561 TGACGTACAT ACATCTGACG TGTGTTTATT TAGACATAAT AGTTATGTTT TCACATCTTT
     1621 TTAATGTTCG CTTAATGCGT ATGCATTCTA GACAATTGTG CTCGGCAACA GTATATTTGT
     1681 GGTGTGCCAA CCAACAACCT GCAGGAGCTC CAGCTTTTGT TCCCTTTAGT GAGGGTTAAT
     1741 TGCGCGCTTG GCGTAATCAT GGTCATAGCT GTTTCCTGTG TGAAATTGTT ATCCGCTCAC
     1801 AATTCCACAC AACATACGAG CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG CCTAATGAGT
     1861 GAGCTAACTC ACATTAATTG CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG GAAACCTGTC
     1921 GTGCCAGCTG CATTAATGAA TCGGCCAACG CGCGGGGAGA GGCGGTTTGC GTATTGGGCG
     1981 CTCTTCCGCT TCCTCGCTCA CTGACTCGCT GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT
     2041 ATCAGCTCAC TCAAAGGCGG TAATACGGTT ATCCACAGAA TCAGGGGATA ACGCAGGAAA
     2101 GAACATGTGA GCAAAAGGCC AGCAAAAGGC CAGGAACCGT AAAAAGGCCG CGTTGCTGGC
     2161 GTTTTTCCAT AGGCTCCGCC CCCCTGACGA GCATCACAAA AATCGACGCT CAAGTCAGAG
     2221 GTGGCGAAAC CCGACAGGAC TATAAAGATA CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT
     2281 GCGCTCTCCT GTTCCGACCC TGCCGCTTAC CGGATACCTG TCCGCCTTTC TCCCTTCGGG
     2341 AAGCGTGGCG CTTTCTCATA GCTCACGCTG TAGGTATCTC AGTTCGGTGT AGGTCGTTCG
     2401 CTCCAAGCTG GGCTGTGTGC ACGAACCCCC CGTTCAGCCC GACCGCTGCG CCTTATCCGG
     2461 TAACTATCGT CTTGAGTCCA ACCCGGTAAG ACACGACTTA TCGCCACTGG CAGCAGCCAC
     2521 TGGTAACAGG ATTAGCAGAG CGAGGTATGT AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG
     2581 GCCTAACTAC GGCTACACTA GAAGGACAGT ATTTGGTATC TGCGCTCTGC TGAAGCCAGT
     2641 TACCTTCGGA AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA CAAACCACCG CTGGTAGCGG
     2701 TGGTTTTTTT GTTTGCAAGC AGCAGATTAC GCGCAGAAAA AAAGGATCTC AAGAAGATCC
     2761 TTTGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT
     2821 GGTCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA AATGAAGTTT
     2881 TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT GCTTAATCAG
     2941 TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT GACTCCCCGT
     3001 CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG CAATGATACC
     3061 GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG CCGGAAGGGC
     3121 CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA ATTGTTGCCG
     3181 GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG CCATTGCTAC
     3241 AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG GTTCCCAACG
     3301 ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC
     3361 TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA TGGCAGCACT
     3421 GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG GTGAGTACTC
     3481 AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC CGGCGTCAAT
     3541 ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG GAAAACGTTC
     3601 TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA TGTAACCCAC
     3661 TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG GGTGAGCAAA
     3721 AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT GTTGAATACT
     3781 CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC TCATGAGCGG
     3841 ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA CATTTCCCCG
     3901 AAAAGTGCCA CCTAAATTGT AAGCGTTAAT ATTTTGTTAA AATTCGCGTT AAATTTTTGT
     3961 TAAATCAGCT CATTTTTTAA CCAATAGGCC GAAATCGGCA AAATCCCTTA TAAATCAAAA
     4021 GAATAGACCG AGATAGGGTT GAGTGTTGTT CCAGTTTGGA ACAAGAGTCC ACTATTAAAG
     4081 AACGTGGACT CCAACGTCAA AGGGCGAAAA ACCGTCTATC AGGGCGATGG CCCACTACGT
     4141 GAACCATCAC CCTAATCAAG TTTTTTGGGG TCGAGGTGCC GTAAAGCACT AAATCGGAAC
     4201 CCTAAAGGGA GCCCCCGATT TAGAGCTTGA CGGGGAAAGC CGGCGAACGT GGCGAGAAAG
     4261 GAAGGGAAGA AAGCGAAAGG AGCGGGCGCT AGGGCGCTGG CAAGTGTAGC GGTCACGCTG
     4321 CGCGTAACCA CCACACCCGC CGCGCTTAAT GCGCCGCTAC AGGGCGCGTC CCATTCGCCA
     4381 TTCAGGCTGC GCAACTGTTG GGAAGGGCGA TCGGTGCGGG CCTCTTCGCT ATTACGCCAG
     4441 CTGGCGAAAG GGGGATGTGC TGCAAGGCGA TTAAGTTGGG TAACGCCAGG GTTTTCCCAG
     4501 TCACGACGTT GTAAAACGAC GGCCAGTGAA TTGTAATACG ACTCACTATA GGGCGAATTG
     4561 GGTACAAGCT GATCT
//
