LOCUS       BC29_(GinB-8GGS-dCas9_N 8065 bp ds-DNA     circular     30-AUG-2016
DEFINITION  .
ACCESSION   urn.local...1wv-5r2m53h                                            
VERSION     urn.local...1wv-5r2m53h                                            
FEATURES             Location/Qualifiers
     promoter        complement(6866..6895)
                     /label="lac prom"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer_bind     6470..6475
                     /label="AgeI"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    2210..2215
                     /label="IG-->RF"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    1805..2230
                     /label="Gin Beta"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    1808..6469
                     /label="GACTACAAAGACCATGACGGTGATTATAAAGATCATGACATCGATTACAAGGATGACGATGACAAGATGGCCCCCAAGAAGAAGAGGAAGGTGGGCATTCACCGCGGGGTACCTGGAGGTTCTATGGGATCCCAACTAGTCAAAAGTGAACTGGAGGAGAAGAAATCTGAACTTCGTCATAAATTGAAATATGTGCCTCATGAATATATTGAATTAATTGAAATTGCCAGAAATTCCACTCAGGATAGAATTCTTGAAATGAAGGTAATGGAATTTTTTATGAAAGTTTATGGATATAGAGGTAAACATTTGGGTGGATCAAGGAAACCGGACGGAGCAATTTATACTGTCGGATCTCCTATTGATTACGGTGTGATCGTGGATACTAAAGCTTATAGCGGAGGTTATAATCTGCCAATTGGCCAAGCAGATGAAATGCAACGATATGTCGAAGAAAATCAAACACGAAACAAACATATCAACCCTAATGAATGGTGGAAAGTCTATCCATCTTCTGTAACGGAATTTAAGTTTTTATTTGTGAGTGGTCACTTTAAAGGAAACTACAAAGCTCAGCTTACACGATTAAATCATATCACTAATTGTAATGGAGCTGTTCTTAGTGTAGAAGAGCTTTTAATTGGTGGAGAAATGATTAAAGCCGGCACATTAACCTTAGAGGAAGTCAGACGGAAATTTAATAACGGCGAGATAAACTTTGGTGGCAGCATGGATAAAAAGTATTCTATTGGTTTAGCTATCGGCACTAATTCCGTTGGATGGGCTGTCATAACCGATGAATACAAAGTACCTTCAAAGAAATTTAAGGTGTTGGGGAACACAGACCGTCATTCGATTAAAAAGAATCTTATCGGTGCCCTCCTATTCGATAGTGGCGAAACGGCAGAGGCGACTCGCCTGAAACGAACCGCTCGGAGAAGGTATACACGTCGCAAGAACCGAATATGTTACTTACAAGAAATTTTTAGCAATGAGATGGCCAAAGTTGACGATTCTTTCTTTCACCGTTTGGAAGAGTCCTTCCTTGTCGAAGAGGACAAGAAACATGAACGGCACCCCATCTTTGGAAACATAGTAGATGAGGTGGCATATCATGAAAAGTACCCAACGATTTATCACCTCAGAAAAAAGCTAGTTGACTCAACTGATAAAGCGGACCTGAGGTTAATCTACTTGGCTCTTGCCCATATGATAAAGTTCCGTGGGCACTTTCTCATTGAGGGTGATCTAAATCCGGACAACTCGGATGTCGACAAACTGTTCATCCAGTTAGTACAAACCTATAATCAGTTGTTTGAAGAGAACCCTATAAATGCAAGTGGCGTGGATGCGAAGGCTATTCTTAGCGCCCGCCTCTCTAAATCCCGACGGCTAGAAAACCTGATCGCACAATTACCCGGAGAGAAGAAAAATGGGTTGTTCGGTAACCTTATAGCGCTCTCACTAGGCCTGACACCAAATTTTAAGTCGAACTTCGACTTAGCTGAAGATGCCAAATTGCAGCTTAGTAAGGACACGTACGATGACGATCTCGACAATCTACTGGCACAAATTGGAGATCAGTATGCGGACTTATTTTTGGCTGCCAAAAACCTTAGCGATGCAATCCTCCTATCTGACATACTGAGAGTTAATACTGAGATTACCAAGGCGCCGTTATCCGCTTCAATGATCAAAAGGTACGATGAACATCACCAAGACTTGACACTTCTCAAGGCCCTAGTCCGTCAGCAACTGCCTGAGAAATATAAGGAAATATTCTTTGATCAGTCGAAAAACGGGTACGCAGGTTATATTGACGGCGGAGCGAGTCAAGAGGAATTCTACAAGTTTATCAAACCCATATTAGAGAAGATGGATGGGACGGAAGAGTTGCTTGTAAAACTCAATCGCGAAGATCTACTGCGAAAGCAGCGGACTTTCGACAACGGTAGCATTCCACATCAAATCCACTTAGGCGAATTGCATGCTATACTTAGAAGGCAGGAGGATTTTTATCCGTTCCTCAAAGACAATCGTGAAAAGAT"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    7166..7192
                     /label="gRNA-for-Esp3I"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    complement(7204..7823)
                     /label="pBR322 origin"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2231..2302
                     /label="8GGS"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    6413..6466
                     /label="NLS"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     promoter        1773..1791
                     /label="T7 prom"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    6404..6412
                     /label="linker"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    2327..2329
                     /label="D10A"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    4817..4819
                     /label="H840A"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    2120..2122
                     /label="H106Y"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    2303..6403
                     /label="dCas9"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     promoter        1671..1752
                     /label="CMV prom"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     promoter        complement(815..843)
                     /label="amp prom"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    1049..1073
                     /label="gRNA-for-Esp3I(2)"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    1149..1758
                     /label="pCMV"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     terminator      6519..6746
                     /label="bGH_PA term"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    complement(7978..773)
                     /label="amp marker"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    complement(814..857)
                     /label="pHU6-rev-Esp3I-rev-2"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(6783..6806)
                     /label="gRNA Esp3I"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
ORIGIN
        1 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
       61 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      121 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      181 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      241 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      301 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      361 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
      421 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      481 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      541 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      601 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      661 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
      721 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
      781 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      841 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      901 cctgacgtcg acggatcggg agatcgatct cccgatcccc tagggtcgac tctcagtaca
      961 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     1021 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     1081 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     1141 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1201 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1261 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1321 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
     1381 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     1441 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1501 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     1561 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     1621 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     1681 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     1741 aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag ggagagccgc
     1801 caccatgctc attggctacg tgcgcgtctc aactaacgac cagaataccg atcttcagag
     1861 gaacgcactg gtttgtgcag gctgcgaaca gattttcgag gacaaactca gcgggacacg
     1921 gacggacaga cctggcctca agcgagcact caagaggctg cagaaaggag acactctggt
     1981 ggtctggaaa ttggaccgcc tgggtcgaag catgaagcat ctcatttctc tggttggcga
     2041 actgcgagaa agggggatca actttcgaag tctgacggat tccatagata caagcagccc
     2101 catgggccgg ttcttcttct acgtgatggg tgcactggct gaaatggaaa gagaactcat
     2161 tatagagcga accatggcag ggcttgcggc tgccaggaat aaaggcaggc ggtttggaag
     2221 accaccaaag ggtggatccg gagggtccgg aggtagtggc ggcagcggtg gttcaggtgg
     2281 cagcggaggg tcaggaggct ctgataaaaa gtattctatt ggtttagcta tcggcactaa
     2341 ttccgttgga tgggctgtca taaccgatga atacaaagta ccttcaaaga aatttaaggt
     2401 gttggggaac acagaccgtc attcgattaa aaagaatctt atcggtgccc tcctattcga
     2461 tagtggcgaa acggcagagg cgactcgcct gaaacgaacc gctcggagaa ggtatacacg
     2521 tcgcaagaac cgaatatgtt acttacaaga aatttttagc aatgagatgg ccaaagttga
     2581 cgattctttc tttcaccgtt tggaagagtc cttccttgtc gaagaggaca agaaacatga
     2641 acggcacccc atctttggaa acatagtaga tgaggtggca tatcatgaaa agtacccaac
     2701 gatttatcac ctcagaaaaa agctagttga ctcaactgat aaagcggacc tgaggttaat
     2761 ctacttggct cttgcccata tgataaagtt ccgtgggcac tttctcattg agggtgatct
     2821 aaatccggac aactcggatg tcgacaaact gttcatccag ttagtacaaa cctataatca
     2881 gttgtttgaa gagaacccta taaatgcaag tggcgtggat gcgaaggcta ttcttagcgc
     2941 ccgcctctct aaatcccgac ggctagaaaa cctgatcgca caattacccg gagagaagaa
     3001 aaatgggttg ttcggtaacc ttatagcgct ctcactaggc ctgacaccaa attttaagtc
     3061 gaacttcgac ttagctgaag atgccaaatt gcagcttagt aaggacacgt acgatgacga
     3121 tctcgacaat ctactggcac aaattggaga tcagtatgcg gacttatttt tggctgccaa
     3181 aaaccttagc gatgcaatcc tcctatctga catactgaga gttaatactg agattaccaa
     3241 ggcgccgtta tccgcttcaa tgatcaaaag gtacgatgaa catcaccaag acttgacact
     3301 tctcaaggcc ctagtccgtc agcaactgcc tgagaaatat aaggaaatat tctttgatca
     3361 gtcgaaaaac gggtacgcag gttatattga cggcggagcg agtcaagagg aattctacaa
     3421 gtttatcaaa cccatattag agaagatgga tgggacggaa gagttgcttg taaaactcaa
     3481 tcgcgaagat ctactgcgaa agcagcggac tttcgacaac ggtagcattc cacatcaaat
     3541 ccacttaggc gaattgcatg ctatacttag aaggcaggag gatttttatc cgttcctcaa
     3601 agacaatcgt gaaaagattg agaaaatcct aacctttcgc ataccttact atgtgggacc
     3661 cctggcccga gggaactctc ggttcgcatg gatgacaaga aagtccgaag aaacgattac
     3721 tccatggaat tttgaggaag ttgtcgataa aggtgcgtca gctcaatcgt tcatcgagag
     3781 gatgaccaac tttgacaaga atttaccgaa cgaaaaagta ttgcctaagc acagtttact
     3841 ttacgagtat ttcacagtgt acaatgaact cacgaaagtt aagtatgtca ctgagggcat
     3901 gcgtaaaccc gcctttctaa gcggagaaca gaagaaagca atagtagatc tgttattcaa
     3961 gaccaaccgc aaagtgacag ttaagcaatt gaaagaggac tactttaaga aaattgaatg
     4021 cttcgattct gtcgagatct ccggggtaga agatcgattt aatgcgtcac ttggtacgta
     4081 tcatgacctc ctaaagataa ttaaagataa ggacttcctg gataacgaag agaatgaaga
     4141 tatcttagaa gatatagtgt tgactcttac cctctttgaa gatcgggaaa tgattgagga
     4201 aagactaaaa acatacgctc acctgttcga cgataaggtt atgaaacagt taaagaggcg
     4261 tcgctatacg ggctggggac gattgtcgcg gaaacttatc aacgggataa gagacaagca
     4321 aagtggtaaa actattctcg attttctaaa gagcgacggc ttcgccaata ggaactttat
     4381 gcagctgatc catgatgact ctttaacctt caaagaggat atacaaaagg cacaggtttc
     4441 cggacaaggg gactcattgc acgaacatat tgcgaatctt gctggttcgc cagccatcaa
     4501 aaagggcata ctccagacag tcaaagtagt ggatgagcta gttaaggtca tgggacgtca
     4561 caaaccggaa aacattgtaa tcgagatggc acgcgaaaat caaacgactc agaaggggca
     4621 aaaaaacagt cgagagcgga tgaagagaat agaagagggt attaaagaac tgggcagcca
     4681 gatcttaaag gagcatcctg tggaaaatac ccaattgcag aacgagaaac tttacctcta
     4741 ttacctacaa aatggaaggg acatgtatgt tgatcaggaa ctggacataa accgtttatc
     4801 tgattacgac gtcgatgcca ttgtacccca atcctttttg aaggacgatt caatcgacaa
     4861 taaagtgctt acacgctcgg ataagaaccg agggaaaagt gacaatgttc caagcgagga
     4921 agtcgtaaag aaaatgaaga actattggcg gcagctccta aatgcgaaac tgataacgca
     4981 aagaaagttc gataacttaa ctaaagctga gaggggtggc ttgtctgaac ttgacaaggc
     5041 cggatttatt aaacgtcagc tcgtggaaac ccgccaaatc acaaagcatg ttgcacagat
     5101 actagattcc cgaatgaata cgaaatacga cgagaacgat aagctgattc gggaagtcaa
     5161 agtaatcact ttaaagtcaa aattggtgtc ggacttcaga aaggattttc aattctataa
     5221 agttagggag ataaataact accaccatgc gcacgacgct tatcttaatg ccgtcgtagg
     5281 gaccgcactc attaagaaat acccgaagct agaaagtgag tttgtgtatg gtgattacaa
     5341 agtttatgac gtccgtaaga tgatcgcgaa aagcgaacag gagataggca aggctacagc
     5401 caaatacttc ttttattcta acattatgaa tttctttaag acggaaatca ctctggcaaa
     5461 cggagagata cgcaaacgac ctttaattga aaccaatggg gagacaggtg aaatcgtatg
     5521 ggataagggc cgggacttcg cgacggtgag aaaagttttg tccatgcccc aagtcaacat
     5581 agtaaagaaa actgaggtgc agaccggagg gttttcaaag gaatcgattc ttccaaaaag
     5641 gaatagtgat aagctcatcg ctcgtaaaaa ggactgggac ccgaaaaagt acggtggctt
     5701 cgatagccct acagttgcct attctgtcct agtagtggca aaagttgaga agggaaaatc
     5761 caagaaactg aagtcagtca aagaattatt ggggataacg attatggagc gctcgtcttt
     5821 tgaaaagaac cccatcgact tccttgaggc gaaaggttac aaggaagtaa aaaaggatct
     5881 cataattaaa ctaccaaagt atagtctgtt tgagttagaa aatggccgaa aacggatgtt
     5941 ggctagcgcc ggagagcttc aaaaggggaa cgaactcgca ctaccgtcta aatacgtgaa
     6001 tttcctgtat ttagcgtccc attacgagaa gttgaaaggt tcacctgaag ataacgaaca
     6061 gaagcaactt tttgttgagc agcacaaaca ttatctcgac gaaatcatag agcaaatttc
     6121 ggaattcagt aagagagtca tcctagctga tgccaatctg gacaaagtat taagcgcata
     6181 caacaagcac agggataaac ccatacgtga gcaggcggaa aatattatcc atttgtttac
     6241 tcttaccaac ctcggcgctc cagccgcatt caagtatttt gacacaacga tagatcgcaa
     6301 acgatacact tctaccaagg aggtgctaga cgcgacactg attcaccaat ccatcacggg
     6361 attatatgaa actcggatag atttgtcaca gcttgggggt gacggtggct ccgattataa
     6421 ggatgatgac gacaagggag gttccccaaa gaagaaaagg aaggtctgaa ccggtcatca
     6481 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     6541 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     6601 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     6661 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     6721 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     6781 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6841 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     6901 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6961 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7021 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7381 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7621 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7861 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8041 tgcctgactc cccgtcgtgt agata
//