LOCUS       pBT279_FERNY-BE4max     8763 bp ds-DNA     circular     22-JAN-2019
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS              642..6005
                      /label="FERNY-BE4max"
                      /ApEinfo_revcolor=#f58a5e
                      /ApEinfo_fwdcolor=#f58a5e
     cloning junction 6049..6052
                      /label="AGCC for BsaI Golden Gate cloning"
                      /ApEinfo_revcolor=#d59687
                      /ApEinfo_fwdcolor=#d59687
     cloning junction 6036..6044
                      /label="USER junction 27"
                      /ApEinfo_revcolor=#ff9ccd
                      /ApEinfo_fwdcolor=#ff9ccd
     cloning junction 699..701
                      /label="AGT for SapI Golden Gate cloning"
                      /ApEinfo_revcolor=#c7b0e3
                      /ApEinfo_fwdcolor=#c7b0e3
     promoter         1..589
                      /label="CMV promoter, full"
                      /ApEinfo_revcolor=#b1ff67
                      /ApEinfo_fwdcolor=#b1ff67
     Promoter         508..589
                      /label="CMV promoter"
                      /ApEinfo_revcolor=#b4abac
                      /ApEinfo_fwdcolor=#b4abac
     misc_feature     627..627
                      /label="TSS"
                      /ApEinfo_revcolor=#d6b295
                      /ApEinfo_fwdcolor=#d6b295
     misc_feature     630..632
                      /label="modest consensus on RRR"
                      /ApEinfo_revcolor=#85dae9
                      /ApEinfo_fwdcolor=#85dae9
     promoter         610..632
                      /label="T7 promoter"
                      /ApEinfo_revcolor=#f58a5e
                      /ApEinfo_fwdcolor=#f58a5e
     NLS              642..698
                      /label="Bipartite SV40 NLS"
                      /ApEinfo_revcolor=#b1ff67
                      /ApEinfo_fwdcolor=#b1ff67
     misc_feature     1024..1030
                      /label="USER jct 10"
                      /ApEinfo_revcolor=#b1ff67
                      /ApEinfo_fwdcolor=#b1ff67
     misc_feature     1008..1010
                      /label="D104"
                      /ApEinfo_revcolor=#84b0dc
                      /ApEinfo_fwdcolor=#84b0dc
     misc_feature     702..1184
                      /label="FERNY"
                      /ApEinfo_revcolor=#75c6a9
                      /ApEinfo_fwdcolor=#75c6a9
     misc_feature     1002..1004
                      /label="H102"
                      /ApEinfo_revcolor=#85dae9
                      /ApEinfo_fwdcolor=#85dae9
     CDS              1185..1280
                      /label="BE4 linker Genscript codons"
                      /ApEinfo_revcolor=#d6b295
                      /ApEinfo_fwdcolor=#d6b295
     cloning junction 1196..1199
                      /label="TAGC for BsaI Golden Gate cloning"
                      /ApEinfo_revcolor=#9eafd2
                      /ApEinfo_fwdcolor=#9eafd2
     cloning junction 1197..1206
                      /label="USER junction 33"
                      /ApEinfo_revcolor=#b7e6d7
                      /ApEinfo_fwdcolor=#b7e6d7
     cloning junction 1188..1190
                      /label="GGC for SapI Golden Gate cloning"
                      /ApEinfo_revcolor=#75c6a9
                      /ApEinfo_fwdcolor=#75c6a9
     misc_feature     1305..1307
                      /label="D10A"
                      /ApEinfo_revcolor=#d6b295
                      /ApEinfo_fwdcolor=#d6b295
     CDS              1281..5381
                      /label="Cas9, Genscript codons"
                      /ApEinfo_revcolor=#d59687
                      /ApEinfo_fwdcolor=#d59687
     source           5382..5951
                      /label="2xUGI Genscript codons"
                      /ApEinfo_revcolor=#b7e6d7
                      /ApEinfo_fwdcolor=#b7e6d7
     NLS              5952..5981
                      /label="SV40 NLS"
                      /ApEinfo_revcolor=#b1ff67
                      /ApEinfo_fwdcolor=#b1ff67
     misc_feature     5982..6002
                      /label="SV40 NLS"
                      /ApEinfo_revcolor=#d59687
                      /ApEinfo_fwdcolor=#d59687
     Terminator       6054..6281
                      /label="bGH PA terminator"
                      /ApEinfo_revcolor=#c7b0e3
                      /ApEinfo_fwdcolor=#c7b0e3
     misc_feature     6570..6576
                      /label="removed SapI site"
                      /ApEinfo_revcolor=#faac61
                      /ApEinfo_fwdcolor=#faac61
     misc_feature     7406..7416
                      /label="USER jct 25"
                      /ApEinfo_revcolor=#f8d3a9
                      /ApEinfo_fwdcolor=#f8d3a9
     misc_feature     6572..6580
                      /label="USER jct 26"
                      /ApEinfo_revcolor=#faac61
                      /ApEinfo_fwdcolor=#faac61
     Marker           complement(7513..8373)
                      /label="bla beta lactamase (ampicillin/carbenicillin resistance)"
                      /ApEinfo_revcolor=#ffef86
                      /ApEinfo_fwdcolor=#ffef86
     Promoter         complement(6401..6430)
                      /label="lac promoter"
                      /ApEinfo_revcolor=#d6b295
                      /ApEinfo_fwdcolor=#d6b295
     misc_feature     7653..7658
                      /label="removed BsaI site"
                      /ApEinfo_revcolor=#b7e6d7
                      /ApEinfo_fwdcolor=#b7e6d7
     Promoter         complement(8415..8443)
                      /label="amp promoter"
                      /ApEinfo_revcolor=#faac61
                      /ApEinfo_fwdcolor=#faac61
     misc_feature     7376..7376
                      /label="G>A"
                      /ApEinfo_revcolor=#d59687
                      /ApEinfo_fwdcolor=#d59687
     origin of replication 6736..7416
                      /label="pUC origin high copy"
                      /ApEinfo_revcolor=#9eafd2
                      /ApEinfo_fwdcolor=#9eafd2
     misc_feature     7656..7664
                      /label="USER jct 31"
                      /ApEinfo_revcolor=#9eafd2
                      /ApEinfo_fwdcolor=#9eafd2
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc
      601 cgcggccgct aatacgactc actataggga gagccgccac catgaaacgg acagccgacg
      661 gaagcgagtt cgagtcacca aagaagaagc ggaaagtcag ttttgagagg aactacgacc
      721 cccgggagct gagaaaggag acatacctgc tgtatgagat caagtggggc aagtccggca
      781 agctgtggag gcactggtgc cagaacaatc gcacacagca cgccgaggtg tacttcctgg
      841 agaacatctt taatgcccgg agattcaatc catctaccca ctgtagcatc acatggtatc
      901 tgagctggtc cccctgcgcc gagtgttctc agaagatcgt ggatttcctg aaggagcacc
      961 ctaacgtgaa tctggagatc tatgtggccc ggctgtacta tcacgaggac gagaggaaca
     1021 ggcAgggccT gcgggatctg gtgaattccg gcgtgaccat cagaatcatg gacctgccag
     1081 attacaacta ttgctggaag accttcgtga gcgatcaggg aggcgacgag gattactggc
     1141 caggacactt cgccccttgg atcaagcagt atagcctgaa gctgtctggc ggatctAgcg
     1201 gaggaTcctc tggcagcgag acaccaggaa caagcgagtc agcaacacca gagagcagtg
     1261 gcggcagcag cggcggcagc gacaagaagt acagcatcgg cctggCcatc ggcaccaact
     1321 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc
     1381 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca
     1441 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac
     1501 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg
     1561 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc
     1621 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca
     1681 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct
     1741 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga
     1801 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc
     1861 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca
     1921 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga
     1981 atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca
     2041 acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc
     2101 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga
     2161 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg
     2221 cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc
     2281 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga
     2341 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt
     2401 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca
     2461 gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc
     2521 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg
     2581 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc
     2641 tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc
     2701 cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga
     2761 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt
     2821 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga
     2881 gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga
     2941 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct
     3001 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc
     3061 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca
     3121 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac
     3181 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga
     3241 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt
     3301 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc
     3361 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg
     3421 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga
     3481 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca
     3541 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga
     3601 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga
     3661 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact
     3721 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg
     3781 actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca
     3841 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg
     3901 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga
     3961 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg
     4021 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc
     4081 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag
     4141 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag
     4201 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa
     4261 ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg
     4321 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca
     4381 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg
     4441 gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg
     4501 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg
     4561 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga
     4621 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg
     4681 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca
     4741 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg
     4801 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga
     4861 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg
     4921 cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact
     4981 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga
     5041 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg
     5101 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca
     5161 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc
     5221 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga
     5281 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc
     5341 tgtacgagac acggatcgac ctgtctcagc tgggaggtga cagcggcggg agcggcggga
     5401 gcggggggag cactaatctg agcgacatca ttgagaagga gactgggaaa cagctggtca
     5461 ttcaggagtc catcctgatg ctgcctgagg aggtggagga agtgatcggc aacaagccag
     5521 agtctgacat cctggtgcac accgcctacg acgagtccac agatgagaat gtgatgctgc
     5581 tgacctctga cgcccccgag tataagcctt gggccctggt catccaggat tctaacggcg
     5641 agaataagat caagatgctg agcggaggat ccggaggatc tggaggcagc accaacctgt
     5701 ctgacatcat cgagaaggag acaggcaagc agctggtcat ccaggagagc atcctgatgc
     5761 tgcccgaaga agtcgaagaa gtgatcggaa acaagcctga gagcgatatc ctggtccata
     5821 ccgcctacga cgagagtacc gacgaaaatg tgatgctgct gacatccgac gccccagagt
     5881 ataagccctg ggctctggtc atccaggatt ccaacggaga gaacaaaatc aaaatgctgt
     5941 ctggcggctc aaaaagaacc gccgacggca gcgaattcga gcccaagaag aagaggaaag
     6001 tctaaccggt catcatcacc atcaccattg agtttAaacc cgcTgatcag cctcgactgt
     6061 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     6121 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     6181 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     6241 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     6301 cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6361 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6421 aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6481 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6541 cgcggggaga ggcggtttgc gtattgggcg cActtccgcT tcctcgctca ctgactcgct
     6601 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6661 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6721 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6781 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6841 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6901 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6961 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7021 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7081 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7141 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagAacagt
     7201 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7261 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7321 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacActca
     7381 gtggaacgaa aactcacgtt aagggAtttt ggtcaTgaga ttatcaaaaa ggatcttcac
     7441 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7501 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7561 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7621 accatctggc cccagtgctg caatgatacc gcgGgAccca cgcTcaccgg ctccagattt
     7681 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7741 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7801 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7861 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7921 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7981 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8041 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8101 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8161 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8221 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8281 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8341 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8401 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8461 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8521 agatcgatct cccgatcccc tagggtcgac tctcagtaca atctgctctg atgccgcata
     8581 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     8641 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     8701 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     8761 gac
//