LOCUS       pCMV-T7-DCV-nCas9-EcKlenow_(JO608)        9918 bp    DNA     circular UNK 24-NOV-2023
DEFINITION  .
ACCESSION   <unknown
KEYWORDS    .
SOURCE      .
  ORGANISM  .
FEATURES             Location/Qualifiers
     CDS             complement(171..1031)
                     /note="pLannotate"
                     /database="addgene"
                     /identity="99.884"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
                     /Color="#777777"
                     /modified_by="lha16"
                     /label="Ampicillin"
     promoter        complement(1032..1136)
                     /note="pLannotate"
                     /database="addgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
                     /label="AmpR promoter"
     enhancer        1420..1785
                     /note="pLannotate"
                     /database="addgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="enhancer"
                     /Color="#bbff33"
                     /modified_by="lha16"
                     /label="CMV enhancer"
     promoter        complement(1469..1497)
                     /created_by="lha16"
                     /modified_by="lha16"
                     /label="AmpR_promoter"
     promoter        1486..1773
                     /created_by="lha16"
                     /modified_by="lha16"
                     /label="CAG_enhancer"
     promoter        1786..2010
                     /note="pLannotate"
                     /database="addgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
                     /modified_by="lha16"
                     /label="CMV promoter"
     promoter        2031..2050
                     /note="pLannotate"
                     /database="addgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
                     /modified_by="lha16"
                     /label="T7 promoter"
     motif           2054..2066
                     /created_by="lha16"
                     /label="Kozak"
     CDS             2063..2422
                     /created_by="joana"
                     /Color="#ffbb33"
                     /modified_by="lha16"
                     /label="DCV HUHe"
     CDS             2423..2494
                     /created_by="lha16"
                     /Color="#ee7700"
                     /modified_by="lha16"
                     /label="GS-XTEN-GS linker"
     CDS             2492..6595
                     /note="pLannotate"
                     /database="addgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
                     /Color="#eeeeee"
                     /modified_by="lha16"
                     /label="nSpCas9(H840A)"
     CDS             5009..5011
                     /created_by="lha16"
                     /label="H840A"
     CDS             6596..6619
                     /created_by="lha16"
                     /Color="#ee7700"
                     /modified_by="lha16"
                     /label="GS linker"
     CDS             6620..6673
                     /created_by="lha16"
                     /Color="#6bba70"
                     /modified_by="lha16"
                     /label="BPNLS"
     CDS             6674..6697
                     /created_by="lha16"
                     /Color="#ee7700"
                     /modified_by="lha16"
                     /label="GS linker"
     CDS             6698..8512
                     /created_by="joana"
                     /Color="#ee0077"
                     /modified_by="lha16"
                     /label="EcKlenow (-exo;D355A/D357A)"
     CDS             8513..8524
                     /created_by="lha16"
                     /Color="#ee7700"
                     /modified_by="lha16"
                     /label="GS linker"
     CDS             8525..8578
                     /created_by="lha16"
                     /Color="#6bba70"
                     /modified_by="lha16"
                     /label="BPNLS"
     CDS             8579..8581
                     /Color="#000000"
                     /created_by="lha16"
                     /label
     terminator      8630..8857
                     /created_by="lha16"
                     /modified_by="lha16"
                     /label="bGH_PA_terminator"
     terminator      9116..9247
                     /created_by="lha16"
                     /label="SV40_PA_terminator"
     rep_origin      complement(9315..16)
                     /note="pLannotate"
                     /database="addgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
                     /modified_by="lha16"
                     /label="pBR322 origin"
ORIGIN      
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gcgaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
     1201 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     1261 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     1321 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     1381 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     1441 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     1501 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1561 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1621 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1681 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     1741 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     1801 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     1861 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     1921 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     1981 agcagagctg gtttagtgaa ccgtcagatc cgctagagat ccgcggccgc taatacgact
     2041 cactataggg agagccgcca ccatggccaa aagcggcaac tacagctaca agcggtgggt
     2101 ctttaccatc aacaacccca ccttcgagga ctacgtgcac gtgctggaat tctgcaccct
     2161 ggataattgc aagttcgcca tcgtgggcga agaaaaggga gccaatggca cacctcacct
     2221 gcagggcttc ctgaacctga gaagcaacgc cagagccgcc gctctggaag agagcctggg
     2281 cggacgggct tggctgagca gagccagagg ctccgatgag gacaacgagg aatactgtgc
     2341 caaggaaagc acctatctcc gggtgggcga gcctgtgtct aagggcagaa gcagcgacct
     2401 ggccgaggcc accagcgctg tgagcggagg ctctagcggc tccgagacac ccggcacatc
     2461 tgagtccgcc acccctgaga gcagcggcgg cagcgacaag aagtacagca tcggcctgga
     2521 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     2581 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     2641 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     2701 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     2761 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     2821 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     2881 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     2941 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     3001 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     3061 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     3121 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     3181 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     3241 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     3301 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     3361 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     3421 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     3481 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     3541 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     3601 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     3661 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     3721 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     3781 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     3841 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     3901 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     3961 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     4021 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     4081 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     4141 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     4201 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     4261 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     4321 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     4381 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     4441 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     4501 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     4561 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     4621 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     4681 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     4741 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     4801 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     4861 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     4921 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     4981 caaccggctg tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga
     5041 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     5101 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     5161 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     5221 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     5281 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     5341 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     5401 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     5461 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     5521 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     5581 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     5641 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     5701 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     5761 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     5821 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     5881 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     5941 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     6001 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     6061 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     6121 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     6181 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     6241 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     6301 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     6361 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     6421 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     6481 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     6541 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactccgg
     6601 cggaagctct ggtggcagca agcggaccgc cgacggctct gaattcgaga gccctaagaa
     6661 gaaaagaaag gtgagcggag gctctagcgg cggaagcgtt atctcctacg acaactacgt
     6721 gaccatcctg gacgaagaga cactgaaggc ttggatcgct aaactggaaa aagcccctgt
     6781 gttcgccttc gccaccgcca ccgactcctt agataacatc agcgccaacc tggtgggcct
     6841 gtctttcgcc atcgagcctg gcgtggccgc ctacatcccc gtggcccacg attacctgga
     6901 tgccccagat caaatcagca gagagagagc actggagctc ctgaagcccc tgctggagga
     6961 cgagaaggcc ctgaaggtgg gacagaacct gaaatacgat aggggaatcc tggctaatta
     7021 cggcatcgag ctgagaggca tcgccttcga taccatgctg gaatcttaca tcctgaacag
     7081 cgtggccggc cgccacgaca tggacagcct ggctgaacgg tggctgaagc acaaaaccat
     7141 caccttcgaa gagatcgccg gaaagggcaa gaaccagctg acatttaatc agatcgcact
     7201 ggaagaagcc ggcagatacg ccgctgagga cgccgacgtg acccttcagc tgcacctgaa
     7261 gatgtggccc gatctgcaga agcacaaggg cccactaaac gtgttcgaga acatcgagat
     7321 gcctctggtg cccgtcctga gcagaatcga gaggaatggc gtgaagatcg accccaaggt
     7381 gctgcacaac cacagcgagg aactgacact gcggctggcc gagctggaaa agaaggccca
     7441 cgaaatcgcc ggcgaggagt tcaacctgtc tagcaccaag cagctacaaa caatcctgtt
     7501 cgagaagcag ggcattaagc ctctgaaaaa gacacctggc ggagccccca gcaccagcga
     7561 agaagtgctg gaggaactgg cgctggatta ccctctgccc aaagtgatcc tggaatatag
     7621 aggactggcc aagctgaagt ctacatacac cgacaagctg cctctgatga tcaaccccaa
     7681 gaccggccgg gtgcatacat cttatcacca ggccgtgaca gccaccggcc ggctgagcag
     7741 cacagatcct aacctgcaaa acatcccagt gcggaacgaa gagggcagaa gaatcagaca
     7801 ggctttcatt gcccctgagg actacgtcat cgtgtctgct gactacagcc agatagaact
     7861 gagaatcatg gcccatctgt ctcgggacaa gggcctgctg accgcttttg cagaaggcaa
     7921 ggacatccac agagccaccg ctgccgaagt gttcggcctg cctctggaaa cagtgaccag
     7981 cgagcaaaga aggtccgcca aggccattaa cttcggcctg atctacggca tgagcgcctt
     8041 tggcctggcc agacagctca acatccctag aaaggaggcc cagaaataca tggacctgta
     8101 cttcgagcgg taccccggcg tgctggagta catggaacgg accagagccc aggctaaaga
     8161 gcagggatac gtggaaaccc tggatggccg gagactgtat ctgcctgaca tcaagagcag
     8221 caacggcgcc agaagagccg ccgctgagag agctgctatc aacgccccta tgcagggcac
     8281 cgccgccgac atcatcaagc gggccatgat cgccgtggac gcctggctgc aggccgagca
     8341 gcctagagtg cggatgatca tgcaggtgca cgacgagctg gtgtttgagg tgcacaagga
     8401 cgacgtggat gctgtggcca agcagattca ccagctgatg gaaaattgca ccagactgga
     8461 cgtgccactg ctggtcgagg ttggaagcgg agagaattgg gaccaggccc actctggcgg
     8521 ctcaaaaaga accgccgacg gcagcgaatt cgagtctccc aagaagaaga ggaaagtctg
     8581 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     8641 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     8701 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     8761 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     8821 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     8881 tggggctcga taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     8941 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     9001 tgtaaagcct agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     9061 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     9121 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     9181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     9241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     9301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     9361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     9421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     9481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     9541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     9601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     9661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     9721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9901 agaaaaaaag gatctcaa
//
