LOCUS       CDS12_Cas9-NLS          6197 bp ds-DNA     circular     18-NOV-2019
DEFINITION  .
KEYWORDS    "Addgene" "OP.kit.position:Plate01-E12" "Type.plasmid:L0"          
            "Description:CDS12_AtcoCas9-NLS(noStop)" "Use:Puchta Arabidopsis
            codon-optimised Cas9 for CRISPR. CDS12 for N-term fusion with a
            CTAG" "Plasmid size, bp:6197" "Bacteria resistance:Cm" "Plant
            resistance" "Phytozome.ID.v3" "Reference:Sauret-Gueto S,
            Frangedakis E, et al (in preparation)" "Primary
            publication:https\://doi.org/10.1111/tpj.12554
            https\://doi.org/10.1371/journal.pone.0205117"
            "Registry.ID.JHLab:HMO_p0_o974" "Name.JHLab"
FEATURES             Location/Qualifiers
     misc_feature    267..4403
                     /label="AtcoCas9 (Puctha) NoStop"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    4406..4409
                     /label="L0 fusion site TTCG"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    259..264
                     /label="BsaI recog site"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     CDS             4652..5311
                     /label="cat / CmR"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    266..269
                     /label="L0 fusion site AATG"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    4383..4403
                     /label="NLS_SV40"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     Replication_ori 5609..6197
                     /label="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(4411..4416)
                     /label="BsaI recog site"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     Terminator      5332..5426
                     /label="lambda t0 terminator"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
ORIGIN
        1 AACGCCAGCA ACGCGGCCTT TTTACGGTTC CTGGCCTTTT GCTGGCCTTT TGCTCACATG
       61 TTCTTTCCTG CGTTATCCCC TGATTCTGTG GATAACCGTA TTACCGCCTT TGAGTGAGCT
      121 GATACCGCTC GCCGCAGCCG AACGACCGAG CGCAGCGAGT CAGTGAGCGA GGAAGCCTGC
      181 ATAACGCGAA GTAATCTTTT CGGTTTTAAA GAAAAAGGGC AGGGTGGTGA CACCTTGCCC
      241 TTTTTTGCCG GACTGCAGgg tctcGAatgg ataagaagta ctctatcgga ctcgatatcg
      301 gaactaactc tgtgggatgg gctgtgatca ccgatgagta caaggtgcca tctaagaagt
      361 tcaaggttct cggaaacacc gataggcact ctatcaagaa aaaccttatc ggtgctctcc
      421 tcttcgattc tggtgaaact gctgaggcta ccagactcaa gagaaccgct agaagaaggt
      481 acaccagaag aaagaacagg atctgctacc tccaagagat cttctctaac gagatggcta
      541 aagtggatga ttcattcttc cacaggctcg aagagtcatt cctcgtggaa gaagataaga
      601 agcacgagag gcaccctatc ttcggaaaca tcgttgatga ggtggcatac cacgagaagt
      661 accctactat ctaccacctc agaaagaagc tcgttgattc tactgataag gctgatctca
      721 ggctcatcta cctcgctctc gctcacatga tcaagttcag aggacacttc ctcatcgagg
      781 gtgatctcaa ccctgataac tctgatgtgg ataagttgtt catccagctc gtgcagacct
      841 acaaccagct tttcgaagag aaccctatca acgcttcagg tgtggatgct aaggctatcc
      901 tctctgctag gctctctaag tcaagaaggc ttgagaacct cattgctcag ctccctggtg
      961 agaagaagaa cggacttttc ggaaacttga tcgctctctc tctcggactc acccctaact
     1021 tcaagtctaa cttcgatctc gctgaggatg caaagctcca gctctcaaag gatacctacg
     1081 atgatgatct cgataacctc ctcgctcaga tcggagatca gtacgctgat ttgttcctcg
     1141 ctgctaagaa cctctctgat gctatcctcc tcagtgatat cctcagagtg aacaccgaga
     1201 tcaccaaggc tccactctca gcttctatga tcaagagata cgatgagcac caccaggatc
     1261 tcacacttct caaggctctt gttagacagc agctcccaga gaagtacaaa gagattttct
     1321 tcgatcagtc taagaacgga tacgctggtt acatcgatgg tggtgcatct caagaagagt
     1381 tctacaagtt catcaagcct atcctcgaga agatggatgg aaccgaggaa ctcctcgtga
     1441 agctcaatag agaggatctt ctcagaaagc agaggacctt cgataacgga tctatccctc
     1501 atcagatcca cctcggagag ttgcacgcta tccttagaag gcaagaggat ttctacccat
     1561 tcctcaagga taacagggaa aagattgaga agattctcac cttcagaatc ccttactacg
     1621 tgggacctct cgctagagga aactcaagat tcgcttggat gaccagaaag tctgaggaaa
     1681 ccatcacccc ttggaacttc gaagaggtgg tggataaggg tgctagtgct cagtctttca
     1741 tcgagaggat gaccaacttc gataagaacc ttccaaacga gaaggtgctc cctaagcact
     1801 ctttgctcta cgagtacttc accgtgtaca acgagttgac caaggttaag tacgtgaccg
     1861 agggaatgag gaagcctgct tttttgtcag gtgagcaaaa gaaggctatc gttgatctct
     1921 tgttcaagac caacagaaag gtgaccgtga agcagctcaa agaggattac ttcaagaaaa
     1981 tcgagtgctt cgattcagtt gagatttctg gtgttgagga taggttcaac gcatctctcg
     2041 gaacctacca cgatctcctc aagatcatta aggataagga tttcttggat aacgaggaaa
     2101 acgaggatat cttggaggat atcgttctta ccctcaccct ctttgaagat agagagatga
     2161 ttgaagaaag gctcaagacc tacgctcatc tcttcgatga taaggtgatg aagcagttga
     2221 agagaagaag atacactggt tggggaaggc tctcaagaaa gctcattaac ggaatcaggg
     2281 ataagcagtc tggaaagaca atccttgatt tcctcaagtc tgatggattc gctaacagaa
     2341 acttcatgca gctcatccac gatgattctc tcacctttaa agaggatatc cagaaggctc
     2401 aggtttcagg acagggtgat agtctccatg agcatatcgc taacctcgct ggatctcctg
     2461 caatcaagaa gggaatcctc cagactgtga aggttgtgga tgagttggtg aaggtgatgg
     2521 gaaggcataa gcctgagaac atcgtgatcg aaatggctag agagaaccag accactcaga
     2581 agggacagaa gaactctagg gaaaggatga agaggatcga ggaaggtatc aaagagcttg
     2641 gatctcagat cctcaaagag caccctgttg agaacactca gctccagaat gagaagctct
     2701 acctctacta cctccagaac ggaagggata tgtatgtgga tcaagagttg gatatcaaca
     2761 ggctctctga ttacgatgtt gatcatatcg tgccacagtc attcttgaag gatgattcta
     2821 tcgataacaa ggtgctcacc aggtctgata agaacagggg taagagtgat aacgtgccaa
     2881 gtgaagaggt tgtgaagaaa atgaagaact attggaggca gctcctcaac gctaagctca
     2941 tcactcagag aaagttcgat aacttgacta aggctgagag gggaggactc tctgaattgg
     3001 ataaggcagg attcatcaag aggcagcttg tggaaaccag gcagatcact aagcacgttg
     3061 cacagatcct cgattctagg atgaacacca agtacgatga gaacgataag ttgatcaggg
     3121 aagtgaaggt tatcaccctc aagtcaaagc tcgtgtctga tttcagaaag gatttccaat
     3181 tctacaaggt gagggaaatc aacaactacc accacgctca cgatgcttac cttaacgctg
     3241 ttgttggaac cgctctcatc aagaagtatc ctaagctcga gtcagagttc gtgtacggtg
     3301 attacaaggt gtacgatgtg aggaagatga tcgctaagtc tgagcaagag atcggaaagg
     3361 ctaccgctaa gtatttcttc tactctaaca tcatgaattt cttcaagacc gagattaccc
     3421 tcgctaacgg tgagatcaga aagaggccac tcatcgagac aaacggtgaa acaggtgaga
     3481 tcgtgtggga taagggaagg gatttcgcta ccgttagaaa ggtgctctct atgccacagg
     3541 tgaacatcgt taagaaaacc gaggtgcaga ccggtggatt ctctaaagag tctatcctcc
     3601 ctaagaggaa ctctgataag ctcattgcta ggaagaagga ttgggaccct aagaaatacg
     3661 gtggtttcga ttctcctacc gtggcttact ctgttctcgt tgtggctaag gttgagaagg
     3721 gaaagagtaa gaagctcaag tctgttaagg aacttctcgg aatcactatc atggaaaggt
     3781 catctttcga gaagaaccca atcgatttcc tcgaggctaa gggatacaaa gaggttaaga
     3841 aggatctcat catcaagctc ccaaagtact cactcttcga actcgagaac ggtagaaaga
     3901 ggatgctcgc ttctgctggt gagcttcaaa agggaaacga gcttgctctc ccatctaagt
     3961 acgttaactt tctttacctc gcttctcact acgagaagtt gaagggatct ccagaagata
     4021 acgagcagaa gcaacttttc gttgagcagc acaagcacta cttggatgag atcatcgagc
     4081 agatctctga gttctctaaa agggtgatcc tcgctgatgc aaacctcgat aaggtgttgt
     4141 ctgcttacaa caagcacaga gataagccta tcagggaaca ggcagagaac atcatccatc
     4201 tcttcaccct taccaacctc ggtgctcctg ctgctttcaa gtacttcgat acaaccatcg
     4261 ataggaagag atacacctct accaaagaag tgctcgatgc taccctcatc catcagtcta
     4321 tcactggact ctacgagact aggatcgatc tctcacagct cggtggtgat tcaagggctg
     4381 atcctaagaa gaagaggaag gttgcTTCGT gagaccGAAT TCCAGAAATC ATCCTTAGCG
     4441 AAAGCTAAGG ATTTTTTTTA TCTGAAATTC TGCCTCGTGA TACGCCTATT TTTATAGGTT
     4501 AATGTCATGA TAATAATGGT TTCTTAGACG TCAGGTGGCA CTCGAGTTGA TCGGGCACGT
     4561 AAGAGGTTCC AACTTTCACC ATAATGAAAT AAGATCACTA CCGGGCGTAT TTTTTGAGTT
     4621 ATCGAGATTT TCAGGAGCTA AGGAAGCTAA AATGGAGAAA AAAATCACTG GATATACCAC
     4681 CGTTGATATA TCCCAATGGC ATCGTAAAGA ACATTTTGAG GCATTTCAGT CAGTTGCTCA
     4741 ATGTACCTAT AACCAGACCG TTCAGCTGGA TATTACGGCC TTTTTAAAGA CCGTAAAGAA
     4801 AAATAAGCAC AAGTTTTATC CGGCCTTTAT TCACATTCTT GCCCGCCTGA TGAATGCTCA
     4861 TCCGGAATTT CGTATGGCAA TGAAAGACGG TGAGCTGGTG ATATGGGATA GTGTTCACCC
     4921 TTGTTACACC GTTTTCCATG AGCAAACTGA AACGTTTTCA TCGCTCTGGA GTGAATACCA
     4981 CGACGATTTC CGGCAGTTTC TACACATATA TTCGCAAGAT GTGGCGTGTT ACGGTGAAAA
     5041 CCTGGCCTAT TTCCCTAAAG GGTTTATTGA GAATATGTTT TTCGTCTCAG CCAATCCCTG
     5101 GGTGAGTTTC ACCAGTTTTG ATTTAAACGT GGCCAATATG GACAACTTCT TCGCCCCCGT
     5161 TTTCACCATG GGCAAATATT ATACGCAAGG CGACAAGGTG CTGATGCCGC TGGCGATTCA
     5221 GGTTCATCAT GCCGTTTGTG ATGGCTTCCA TGTCGGCAGA ATGCTTAATG AATTACAACA
     5281 GTACTGCGAT GAGTGGCAGG GCGGGGCGTA ATTTGATATC GAGCTCGCTT GGACTCCTGT
     5341 TGATAGATCC AGTAATGACC TCAGAACTCC ATCTGGATTT GTTCAGAACG CTCGGTTGCC
     5401 GCCGGGCGTT TTTTATTGGT GAGAATCCAA GCCTCGAGCT GTCAGACCAA GTTTACTCAT
     5461 ATATACTTTA GATTGATTTA AAACTTCATT TTTAATTTAA AAGGATCTAG GTGAAGATCC
     5521 TTTTTGATAA TCTCATGACC AAAATCCCTT AACGTGAGTT TTCGTTCCAC TGAGCGTCAG
     5581 ACCCCGTAGA AAAGATCAAA GGATCTTCTT GAGATCCTTT TTTTCTGCGC GTAATCTGCT
     5641 GCTTGCAAAC AAAAAAACCA CCGCTACCAG CGGTGGTTTG TTTGCCGGAT CAAGAGCTAC
     5701 CAACTCTTTT TCCGAAGGTA ACTGGCTTCA GCAGAGCGCA GATACCAAAT ACTGTTCTTC
     5761 TAGTGTAGCC GTAGTTAGGC CACCACTTCA AGAACTCTGT AGCACCGCCT ACATACCTCG
     5821 CTCTGCTAAT CCTGTTACCA GTGGCTGCTG CCAGTGGCGA TAAGTCGTGT CTTACCGGGT
     5881 TGGACTCAAG ACGATAGTTA CCGGATAAGG CGCAGCGGTC GGGCTGAACG GGGGGTTCGT
     5941 GCACACAGCC CAGCTTGGAG CGAACGACCT ACACCGAACT GAGATACCTA CAGCGTGAGC
     6001 TATGAGAAAG CGCCACGCTT CCCGAAGGGA GAAAGGCGGA CAGGTATCCG GTAAGCGGCA
     6061 GGGTCGGAAC AGGAGAGCGC ACGAGGGAGC TTCCAGGGGG AAACGCCTGG TATCTTTATA
     6121 GTCCTGTCGG GTTTCGCCAC CTCTGACTTG AGCGTCGATT TTTGTGATGC TCGTCAGGGG
     6181 GGCGGAGCCT GTGGAAA
//