LOCUS       pCMV-T7-TraI-nCas9-EcKlenow_(JO604)       10548 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"
                     /modified_by="lha16"
                     /label="AmpR promoter"
     enhancer        1420..1785
                     /note="pLannotate"
                     /database="addgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="enhancer"
                     /label="CMV enhancer"
     enhancer        complement(1469..1497)
                     /created_by="lha16"
                     /modified_by="lha16"
                     /label="AmpR_promoter"
     enhancer        1486..1773
                     /created_by="lha16"
                     /label="CAG_enhancer"
     promoter        1786..1989
                     /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"
                     /label="T7 promoter"
     CDS             2063..3052
                     /created_by="joana"
                     /Color="#ffbb33"
                     /modified_by="lha16"
                     /label="TraI HUHe"
     CDS             3053..3124
                     /Color="#ff7400"
                     /created_by="lha16"
                     /label="GS-XTEN-GS linker"
     CDS             3122..7225
                     /note="pLannotate"
                     /database="addgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
                     /Color="#eeeeee"
                     /modified_by="lha16"
                     /label="nSpCas9(H840A)"
     CDS             5639..5641
                     /created_by="lha16"
                     /label="H840A"
     CDS             7226..7249
                     /Color="#ee7700"
                     /created_by="lha16"
                     /label="GS linker"
     CDS             7250..7303
                     /Color="#6bba70"
                     /created_by="lha16"
                     /label="BPNLS"
     CDS             7304..7327
                     /created_by="lha16"
                     /Color="#ff7400"
                     /modified_by="lha16"
                     /label="GS linker"
     CDS             7328..9142
                     /created_by="joana"
                     /Color="#ee0077"
                     /modified_by="lha16"
                     /label="EcKlenow(exo-;D355A/E375A)"
     CDS             9143..9154
                     /Color="#ee7700"
                     /created_by="lha16"
                     /label="GS linker"
     CDS             9155..9208
                     /Color="#6bba70"
                     /created_by="lha16"
                     /label="BPNLS"
     CDS             9209..9211
                     /Color="#000000"
                     /created_by="lha16"
                     /label
     terminator      9260..9487
                     /created_by="lha16"
                     /label="bGH_PA_terminator"
     terminator      9746..9877
                     /created_by="lha16"
                     /label="SV40_PA_terminator"
     rep_origin      complement(9945..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 ccatgatgag catcgcccag gtcagaagcg ctggctccgc
     2101 cggcaattac tacaccgaca aagataacta ctacgtgctg ggctccatgg gcgaacggtg
     2161 ggccggccgt ggagctgaac agctgggcct gcagggctct gtggataagg acgtgttcac
     2221 caggctgctg gaaggcagac ttcctgatgg cgccgacctg agcagaatgc aagacggcag
     2281 caaccggcac agacctggct acgacctgac cttcagcgcc cctaagagcg tgtccatgat
     2341 ggctatgctg ggcggagata agcggctgat cgacgcccac aaccaggctg ttgacttcgc
     2401 cgtgcggcag gtggaagcct tggctagcac cagagtgatg accgacggcc agagcgagac
     2461 agtgctgacc ggcaacctgg tgatggccct cttcaaccac gacacaagcc gggatcagga
     2521 gcctcagctg cacacccacg ccgtggtcgc caacgtgacc cagcacaacg gcgagtggaa
     2581 gacactgagc agcgacaagg tgggaaaaac cggattcatc gagaacgtgt acgccaatca
     2641 aatcgccttc ggcagactgt acagagagaa gctgaaagag caggtggaag ccctgggata
     2701 cgaaaccgag gtggtgggca agcacggcat gtgggagatg cctggcgtgc ccgtggaagc
     2761 ttttagcggc agatctcaga caatccggga agccgtcgga gaggacgcca gcctgaagtc
     2821 cagagatgtg gccgctctgg acacccgcaa gtctaagcag catgtggacc ccgagattaa
     2881 gatggccgag tggatgcaga ccctgaagga aaccggcttt gacatcagag cctatagaga
     2941 tgccgccgac cagcgggccg atctgagaac cctgacaccc ggacctgcca gccaggacgg
     3001 ccccgacgtg cagcaagccg tgacacaggc catcgccgga ctgtccgaga gaagcggcgg
     3061 ctctagcggc agcgagacac ctggtacaag cgaatctgca accccagaga gcagcggcgg
     3121 ctctgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     3181 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     3241 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     3301 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     3361 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     3421 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     3481 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     3541 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     3601 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     3661 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     3721 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     3781 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     3841 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     3901 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     3961 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     4021 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     4081 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     4141 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     4201 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     4261 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     4321 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     4381 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     4441 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     4501 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     4561 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     4621 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     4681 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     4741 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     4801 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     4861 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     4921 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     4981 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     5041 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     5101 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     5161 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     5221 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     5281 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     5341 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     5401 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     5461 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     5521 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     5581 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc
     5641 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     5701 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     5761 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     5821 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     5881 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     5941 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     6001 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     6061 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     6121 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     6181 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     6241 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     6301 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     6361 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     6421 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     6481 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     6541 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     6601 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     6661 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     6721 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     6781 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     6841 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     6901 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     6961 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     7021 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     7081 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     7141 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     7201 cgacctgtct cagctgggag gtgactccgg cggaagctct ggtggcagca agcggaccgc
     7261 cgacggctct gaattcgaga gccctaagaa gaaaagaaag gtgagcggag gctctagcgg
     7321 cggaagcgtt atctcctacg acaactacgt gaccatcctg gacgaagaga cactgaaggc
     7381 ttggatcgct aaactggaaa aagcccctgt gttcgccttc gccaccgcca ccgactcctt
     7441 agataacatc agcgccaacc tggtgggcct gtctttcgcc atcgagcctg gcgtggccgc
     7501 ctacatcccc gtggcccacg attacctgga tgccccagat caaatcagca gagagagagc
     7561 actggagctc ctgaagcccc tgctggagga cgagaaggcc ctgaaggtgg gacagaacct
     7621 gaaatacgat aggggaatcc tggctaatta cggcatcgag ctgagaggca tcgccttcga
     7681 taccatgctg gaatcttaca tcctgaacag cgtggccggc cgccacgaca tggacagcct
     7741 ggctgaacgg tggctgaagc acaaaaccat caccttcgaa gagatcgccg gaaagggcaa
     7801 gaaccagctg acatttaatc agatcgcact ggaagaagcc ggcagatacg ccgctgagga
     7861 cgccgacgtg acccttcagc tgcacctgaa gatgtggccc gatctgcaga agcacaaggg
     7921 cccactaaac gtgttcgaga acatcgagat gcctctggtg cccgtcctga gcagaatcga
     7981 gaggaatggc gtgaagatcg accccaaggt gctgcacaac cacagcgagg aactgacact
     8041 gcggctggcc gagctggaaa agaaggccca cgaaatcgcc ggcgaggagt tcaacctgtc
     8101 tagcaccaag cagctacaaa caatcctgtt cgagaagcag ggcattaagc ctctgaaaaa
     8161 gacacctggc ggagccccca gcaccagcga agaagtgctg gaggaactgg cgctggatta
     8221 ccctctgccc aaagtgatcc tggaatatag aggactggcc aagctgaagt ctacatacac
     8281 cgacaagctg cctctgatga tcaaccccaa gaccggccgg gtgcatacat cttatcacca
     8341 ggccgtgaca gccaccggcc ggctgagcag cacagatcct aacctgcaaa acatcccagt
     8401 gcggaacgaa gagggcagaa gaatcagaca ggctttcatt gcccctgagg actacgtcat
     8461 cgtgtctgct gactacagcc agatagaact gagaatcatg gcccatctgt ctcgggacaa
     8521 gggcctgctg accgcttttg cagaaggcaa ggacatccac agagccaccg ctgccgaagt
     8581 gttcggcctg cctctggaaa cagtgaccag cgagcaaaga aggtccgcca aggccattaa
     8641 cttcggcctg atctacggca tgagcgcctt tggcctggcc agacagctca acatccctag
     8701 aaaggaggcc cagaaataca tggacctgta cttcgagcgg taccccggcg tgctggagta
     8761 catggaacgg accagagccc aggctaaaga gcagggatac gtggaaaccc tggatggccg
     8821 gagactgtat ctgcctgaca tcaagagcag caacggcgcc agaagagccg ccgctgagag
     8881 agctgctatc aacgccccta tgcagggcac cgccgccgac atcatcaagc gggccatgat
     8941 cgccgtggac gcctggctgc aggccgagca gcctagagtg cggatgatca tgcaggtgca
     9001 cgacgagctg gtgtttgagg tgcacaagga cgacgtggat gctgtggcca agcagattca
     9061 ccagctgatg gaaaattgca ccagactgga cgtgccactg ctggtcgagg ttggaagcgg
     9121 agagaattgg gaccaggccc actctggcgg ctcaaaaaga accgccgacg gcagcgaatt
     9181 cgagtctccc aagaagaaga ggaaagtctg accggtcatc atcaccatca ccattgagtt
     9241 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     9301 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     9361 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     9421 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     9481 tctatggctt ctgaggcgga aagaaccagc tggggctcga taccgtcgac ctctagctag
     9541 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     9601 ccacacaaca tacgagccgg aagcataaag tgtaaagcct agggtgccta atgagtgagc
     9661 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     9721 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     9781 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     9841 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     9901 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     9961 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
    10021 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
    10081 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
    10141 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
    10201 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
    10261 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
    10321 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
    10381 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
    10441 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
    10501 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaa
//
