LOCUS       Exported                8745 bp ds-DNA     circular UNA 12-OCT-2022
DEFINITION  natural circular DNA
ACCESSION   urn.local...6s-823ng4u
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8745)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 13, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8745
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     promoter        104..680
                     /label=pCMV immearly protomter
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    183..351
                     /label=CAG enhancer
                     /note="/created_by=User"
     misc_feature    308
                     /label=C
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=C"
     promoter        638..707
                     /label=CMV promoter
                     /note="/created_by=User"
     promoter        728..746
                     /label=T7 protomet
                     /note="/created_by=User"
     CDS             760..1260
                     /label=TadAC1.14
                     /note="/created_by=User"
                     /note="/modified_by=ngaudelli"
     misc_feature    762^763
                     /label=TC
                     /note="Geneious type: Editing History Deletion"
                     /note="/Original_Bases=TC"
     misc_feature    763..765
                     /label=~S2H
                     /note="/created_by=dlam"
     misc_feature    764..765
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    826..828
                     /label=W23R
                     /note="/created_by=ngaudelli"
     misc_feature    865..867
                     /label=H36L
                     /note="/created_by=User"
     misc_feature    901..903
                     /label=P48A
                     /note="/created_by=ngaudelli"
                     /note="/modified_by=ngaudelli"
     misc_feature    904..906
                     /label=~I49K
                     /note="/created_by=dlam"
     misc_feature    905..906
                     /label=TC
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=TC"
     misc_feature    910..912
                     /label=R51L
                     /note="/created_by=User"
     misc_feature    963^964
                     /label=GTGATGCAGAATTATCGACTT
                     /note="Geneious type: Editing History Deletion"
                     /note="/Original_Bases=GTGATGCAGAATTATCGACTT"
     misc_feature    984^985
                     /label=T
                     /note="Geneious type: Editing History Deletion"
                     /note="/Original_Bases=T"
     misc_feature    985..987
                     /label=~I76
                     /note="/created_by=dlam"
     misc_feature    987
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    1003..1005
                     /label=V82S
                     /note="/created_by=nicolegaudelli"
     misc_feature    1009..1011
                     /label=L84F
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    1075..1077
                     /label=A106V
                     /note="/created_by=User"
     misc_feature    1081..1083
                     /label=D108N
                     /note="/created_by=User"
     misc_feature    1093..1095
                     /label=~G112H
                     /note="/created_by=dlam"
                     /note="/modified_by=dlam"
     misc_feature    1126..1128
                     /label=Y123H
                     /note="/created_by=nicolegaudelli"
     misc_feature    1195..1197
                     /label=S146C
                     /note="/created_by=User"
     misc_feature    1198..1221
                     /label=ACTTTTTTTCGCATGCCCAGGCAG
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=ACTTTTTTTCGCATGCCCAGGCAG"
     misc_feature    1198..1200
                     /label=Y147R
                     /note="/created_by=nicolegaudelli"
     misc_feature    1213..1215
                     /label=R152P
                     /note="/created_by=ngaudelli"
     misc_feature    1219..1221
                     /label=Q154R
                     /note="/created_by=nicolegaudelli"
     misc_feature    1222..1224
                     /label=E155V
                     /note="/created_by=User"
     misc_feature    1225..1227
                     /label=I156F
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    1228..1230
                     /label=K157N
                     /note="/created_by=User"
     CDS             1261..1356
                     /label=linker
                     /note="/created_by=nicolegaudelli"
     CDS             1285..1332
                     /label=x ten
                     
                     /note="/created_by=User"
     CDS             1357..5457
                     /label=nCas9 (D10A)
                     /note="hSpCas9"
                     /note="/modified_by=ngaudelli"
     CDS             5458..6015
                     /label=UGI
                     /note="/modified_by=dlam"
     CDS             6016..6105
                     /label=linker+SV40 NLS
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="/modified_by=dlam"
     terminator      complement(6154..6381)
                     /label=bgH terminator
                     /note="/created_by=User"
     promoter        complement(6501..6530)
                     /label=lac promoter
                     /note="/created_by=User"
     rep_origin      6839..7458
                     /label=pBR322
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    7050
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    7295
                     /label=G
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=G"
     CDS             complement(7613..8473)
                     /label=beta-lactamase
                     /note="/created_by=User"
     promoter        complement(8515..8543)
                     /label=pAmpR
                     /note="/created_by=User"
                     /note="/modified_by=User"
ORIGIN
        1 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
       61 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      121 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      181 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      241 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      481 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      541 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      601 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      661 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
      721 cggccgctaa tacgactcac tatagggaga gccgccacca tgcatgaagt cgagttttcc
      781 catgagtact ggatgagaca cgcattgact ctcgcaaaga gggctcgaga tgaacgcgag
      841 gtgcccgtgg gggcagtact cgtgctcaac aatcgcgtaa tcggcgaagg ttggaatagg
      901 gcaaagggac tccacgaccc cactgcacat gcggaaatca tggcccttcg acagggaggg
      961 cttgtgatgc agaattatcg acttatcgat gcgacgctgt actcgacgtt tgaaccttgc
     1021 gtaatgtgcg cgggagctat gattcactcc cgcattggac gagttgtatt cggtgttcgc
     1081 aacgccaaga cgcatgccgc aggttcactg atggacgtgc tgcatcatcc aggcatgaac
     1141 caccgggtag aaatcacaga aggcatattg gcggacgaat gtgcggcgct gttgtgtcgt
     1201 ttttttcgca tgcccaggcg tgtctttaac gcccagaaaa aagcacaatc ctctactgac
     1261 tctggtggtt cttctggtgg ttctagcggc agcgagactc ccgggacctc agagtccgcc
     1321 acacccgaaa gttctggtgg ttcttctggt ggttctgaca agaagtacag catcggcctg
     1381 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     1441 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     1501 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     1561 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     1621 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     1681 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     1741 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     1801 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     1861 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     1921 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     1981 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     2041 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     2101 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     2161 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     2221 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     2281 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     2341 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     2401 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     2461 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     2521 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     2581 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     2641 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     2701 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     2761 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     2821 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     2881 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     2941 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     3001 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     3061 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     3121 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     3181 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     3241 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     3301 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     3361 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     3421 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     3481 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     3541 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     3601 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     3661 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     3721 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     3781 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     3841 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
     3901 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     3961 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     4021 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     4081 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     4141 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     4201 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     4261 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     4321 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     4381 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     4441 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     4501 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     4561 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     4621 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     4681 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     4741 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     4801 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     4861 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     4921 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     4981 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
     5041 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     5101 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     5161 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     5221 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     5281 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     5341 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     5401 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggtgactct
     5461 ggtggaagcg gaggatctgg cggcagcacc aatctgagcg acatcatcga gaaagagaca
     5521 ggcaagcagc tggtcatcca agagtccatc ctgatgctgc ctgaagaggt ggaagaagtg
     5581 atcggcaaca agcccgagtc cgacatcctg gtgcacaccg cctacgatga gagcaccgac
     5641 gagaacgtga tgctgctgac ctctgacgcc cctgagtaca agccttgggc tctcgtgatc
     5701 caggacagca acggcgagaa caagatcaag atgctgagcg gcggctctgg tggctctggc
     5761 ggatctacaa acctgtccga tattattgag aaagaaaccg ggaaacagct cgtgattcaa
     5821 gagtctattc tcatgctccc ggaagaagtc gaggaagtca ttggaaacaa gcctgagagc
     5881 gatattctgg tccatacagc ctacgacgag tctaccgatg agaatgtcat gctcctcacc
     5941 agcgacgctc ccgagtataa gccatgggca cttgtcattc aggactccaa tggggaaaac
     6001 aaaatcaaaa tgctcccaaa gaaaaaacgc aaggtggagg gagctgataa gcgcaccgcc
     6061 gatggttccg agttcgaaag ccccaagaag aagaggaaag tctaaccggt catcatcacc
     6121 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     6181 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     6241 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     6301 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     6361 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt
     6421 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6481 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctagggtg
     6541 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6601 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6661 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6721 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6781 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6841 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6901 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6961 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7021 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7081 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7141 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7201 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7261 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7321 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7381 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7441 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7501 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7561 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7621 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7681 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7741 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7801 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7861 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7921 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7981 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8041 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8101 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8161 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8221 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8281 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8341 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8401 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8461 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8521 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8581 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
     8641 tagggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     8701 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aattt
//
