LOCUS       Lac-BZ-Large-HEPN-BsaI  8098 bp ds-DNA     circular     21-MAR-2017
DEFINITION  .
KEYWORDS    "accession:addgene_65823_111984"                                   
FEATURES             Location/Qualifiers
     misc_feature    3431..7102
                     /label="BZ large protein"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    7188..7196
                     /label="spacer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    7197..7202
                     /label="BsaI"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    7204..7239
                     /label="BZ DR"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        7106..7140
                     /label="J23119 promoter"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    3357..3386
                     /label="Gibson homology EcoRV-pACYC"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    3776..3793
                     /label="HEPN1"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    7240..7269
                     /label="Gibson homology R EcoRV pACYC"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    7183..7187
                     /label="BsaI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     promoter        3387..3416
                     /label="Lac promoter"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    7146..7181
                     /label="BZ DR"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    6959..6976
                     /label="HEPN2"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     RBS             3417..3422
                     /label="Shine-Dalgarno sequence"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
ORIGIN
        1 aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg
       61 cggctcttac cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg
      121 cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct
      181 gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg
      241 gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
      301 aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
      361 cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
      421 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
      481 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
      541 aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcccc
      601 tacgtgctgc tgaagttgcc cgcaacagag agtggaacca accggtgata ccacgatact
      661 atgactgaga gtcaacgcca tgagcggcct catttcttat tctgagttac aacagtccgc
      721 accgctgtcc ggtagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
      781 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgccca acagtccccc
      841 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ccctgcacca ttatgttccg
      901 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
      961 gcgctaaccg tttttatcag gctctgggag gcagaataaa tgatcatatc gtcaattatt
     1021 acctccacgg ggagagcctg agcaaactgg cctcaggcat ttgagaagca cacggtcaca
     1081 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     1141 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     1201 tattatcact tattcaggcg tagcaccagg cgtttaaggg caccaataac tgccttaaaa
     1261 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     1321 gacatggaag ccatcacaga cggcatgatg aacctgaatc gccagcggca tcagcacctt
     1381 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     1441 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     1501 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     1561 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     1621 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     1681 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     1741 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     1801 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     1861 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     1921 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     1981 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     2041 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     2101 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     2161 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     2221 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     2281 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     2341 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     2401 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     2461 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     2521 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     2581 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     2641 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     2701 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     2761 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     2821 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     2881 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     2941 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     3001 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     3061 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     3121 aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     3181 catacgatat aagttgtaat tctcatgttt gacagcttat catcgataag ctttaatgcg
     3241 gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg
     3301 cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc
     3361 cggtactgcc gggcctcttg cgggatTTTA CACTTTATGC TTCCGGCTCG TATGTTAGGA
     3421 GGTCTTTATC atggaaaaca aaacatcttt ggggaataat atttattaca acccttttaa
     3481 accacaagat aaatcgtatt ttgcgggtta tttcaatgca gcaatggaaa atacagatag
     3541 tgtatttaga gaattgggaa aaagattaaa aggtaaagag tacacttcgg agaatttttt
     3601 tgatgctatc tttaaggaaa atatttcatt agtggaatac gaaaggtatg tgaaattgct
     3661 ttctgattat ttccctatgg ctcgtctttt ggataaaaaa gaagttccta taaaagaacg
     3721 aaaagaaaat tttaaaaaga attttaaagg cattataaaa gccgtaagag atttaGCAaa
     3781 cttttatacg GCAaaggaac acggagaggt agaaattaca gatgaaatat ttggcgtatt
     3841 ggatgagatg ctgaaaagca cggttttaac ggtaaaaaag aaaaaagtaa aaaccgataa
     3901 aacaaaagaa atcctgaaaa aaagcataga aaaacaattg gatattttat gccaaaagaa
     3961 attagaatat ctaagagaca cagcaagaaa aatagaggag aagcgtagaa atcaaagaga
     4021 aagaggagag aaagaattgg tagctccatt caaatacagt gataaaaggg atgatttaaT
     4081 AGCAGCGATT TATAATGATG CTTTTGATGt ttatatcgat aagaaaaaag atagcctaaa
     4141 agaatccagc aaagcaaagt acaataccaa aagtgatcct cagcaagaag aaggcgattt
     4201 gaaaattcca atttcgaaaa acggagtagt cttcctgctt tctttgtttc tcaccaaaca
     4261 agaaatacac gcattcaaat ctaaaatagc agggtttaag gctacggtta tcgatgaagc
     4321 aacggtatct gaagcaacgg tgagccatgg aaaaaacagc atttgtttta tggctacgca
     4381 cgaaattttc tcgcatttgg cttacaaaaa attgaaacga aaagtgagaa cggctgaaat
     4441 caactatggt gaggcagaaa atgctgaaca gctttccgtt tatgccaaag aaactctaat
     4501 gatgcagatg cttgatgaat tgagcaaagt acccgatgtg gtctatcaga atttgagcga
     4561 agatgtacaa aaaactttca tcgaagattg gaatgaatac ctaaaagaga acaatggcga
     4621 tgtagggaca atggaagaag agcaggtaat tcatccagtg attcgcaagc gttacgaaga
     4681 taaattcaat tattttgcca ttcgattttt agatgagttt gctcagttcc ctacgcttcg
     4741 ttttcaagtg catttaggga attatcttca cgattctcgc cctaaagaaa atctgatttc
     4801 tgaccgaaga ataaaagaaa aaatcaccgt tttcggaagg ctttcggaat tggaacataa
     4861 aaaagcattg tttataaaaa ataccgaaac caatgaagat agagaacact attgggaaat
     4921 ttttcctaac cccaattatg attttcctaa ggaaaatatt tccgttaatg ataaagattt
     4981 tcctatcgcg ggtagtattt tggacagaga gaaacagcct gtcgcaggga aaattggtat
     5041 aaaagtgaaa ttgcttaacc aacaatatgt ttcagaagta gataaagccg taaaagccca
     5101 tcagctcaag caaagaaaag ccagcaagcc gtctatccag aacattatag aagaaatagt
     5161 gccaatcaat gaaagcaatc caaaagaagc tattgttttc ggtgggcaac ccacggctta
     5221 tctgagtatg aatgacattc attccatttt gtatgagttt ttcgataaat gggagaaaaa
     5281 gaaagaaaag ttagaaaaga aaggtgaaaa agaattaaga aaggaaattg gaaaagagtt
     5341 agaaaagaaa atcgtaggaa aaattcaagc tcaaatccag caaatcatag ataaagatac
     5401 caatgccaag atattaaaac cttatcaaga cggaaattct actgctattg ataaagagaa
     5461 attgataaaa gatttgaagc aagagcaaaa cattctacaa aaattaaaag atgaacaaac
     5521 ggttcgtgaa aaagaatata atgattttat tgcttatcaa gataaaaata gggagataaa
     5581 taaagtcaga gatagaaatc ataaacaata tttaaaagat aaccttaaac gaaagtaccc
     5641 tgaagctcct gcacgaaaag aagttcttta ttatcgagaa aaaggaaaag tagcggtttg
     5701 gttggcgaat gacatcaaac gctttatgcc taccgatttc aaaaacgaat ggaaagggga
     5761 gcaacacagc ttgttacaaa aatcgttggc gtattatgag caatgtaaag aggagttgaa
     5821 aaatcttttg cctgaaaaag tatttcagca tttacctttt aagttgggag gatattttca
     5881 acaaaaatat ttgtaccaat tttacacttg ttatttagat aaacggttgg aatacatcag
     5941 tgggttggtg caacaagccg aaaactttaa atccgaaaat aaagtcttca aaaaggtaga
     6001 aaatgagtgt tttaaatttt tgaaaaaaca aaattatact cacaaagaat tagatgctcg
     6061 ggttcagtct atattggggt atcctatctt tttagagcgg ggctttatgg atgaaaaacc
     6121 tacgataatc aaaggaaaaa cttttaaagg aaatgaagct ctttttgccg attggtttag
     6181 gtattacaag gaatatcaga attttcagac attttatgat actgaaaatt atccgcttgt
     6241 agaattggaa aaaaaacaag cagaccgaaa acgaaaaact aaaatctacc aacaaaagaa
     6301 aaacgatgtg tttacacttt tgatggcaaa acatattttc aaatcggtat tcaagcagga
     6361 tagcatcgat cagttttcgt tagaagattt gtatcaatct cgggaggaac gcttaggaaa
     6421 tcaggagaga gccagacaaa cgggcgagcg aaatactaac tatatttgga acaaaaccgt
     6481 tgatttaaag ttgtgtgacg gaaaaattac ggtagaaaat gtaaagctaa aaaatgtagg
     6541 agatttcata aaatatgaat acgaccaaag ggttcaagct ttcttaaaat atgaggaaaa
     6601 tatagaatgg caggcgtttc taatcaaaga gagtaaggaa gaggaaaatt atccgtatgt
     6661 ggtggaaaga gaaatagagc aatatgaaaa agtgagaagg gaagaactgt tgaaggaagt
     6721 ccaccttata gaggaatata ttttggaaaa ggtaaaagat aaggaaatat taaaaaaagg
     6781 ggataatcaa aatttcaaat actacattct caacggactt ttaaagcaac ttaaaaatga
     6841 agatgtggag agctataagg tattcaactt aaatactgaa cctgaagatg tgaacattaa
     6901 ccaactaaaa caagaagcaa ccgatttaga acaaaaagct tttgttttaa cctatattGC
     6961 Aaataaattt gcaGCAaacc aattacctaa aaaagaattt tgggactatt gtcaagaaaa
     7021 atacggcaaa atagaaaaag aaaaaaccta tgccgagtat tttgctgagg tatttaaaaa
     7081 ggaaaaagaa gcactgataa agTAATTGAC AGCTAGCTCA GTCCTAGGTA TAATGCTAGC
     7141 GCTGAGTTGG AACTGCTCTC ATTTTGGAGG GTAATCACAA CGAGACCATG GCGGAAGGTC
     7201 TCAgttggaa ctgctctcat tttggagggt aatcacaaca tcgtccattc cgacagcatc
     7261 gccagtcact atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc
     7321 gttctcggag cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt
     7381 ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat cctctacgcc
     7441 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     7501 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     7561 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     7621 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     7681 caggagtcgc ataagggaga gcgtcgaccg atgcccttga gagccttcaa cccagtcagc
     7741 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     7801 atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt
     7861 cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc
     7921 gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc
     7981 gccggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg
     8041 atggccttcc ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgc
//