LOCUS       pCMV-T7-PCV2-nSaCas9-EcKlenow_(MLE194)        8967 bp    DNA     circular SYN 08-APR-2024
DEFINITION  .
ACCESSION
VERSION     .
KEYWORDS    .
SOURCE      .
  ORGANISM  .
COMMENT     Annotated with pLannotate v1.2.0.
FEATURES             Location/Qualifiers
     promoter        complement(113..217)
                     /note="pLannotate"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
                     /modified_by="lha16"
                     /label="AmpR promoter"
     promoter        501..866
                     /created_by="lha16"
                     /label="CMV Enhancer"
     promoter        complement(550..578)
                     /created_by="lha16"
                     /label="AmpR_promoter"
     promoter        567..854
                     /created_by="lha16"
                     /label="CAG_enhancer"
     promoter        867..1091
                     /created_by="lha16"
                     /label="CMV_promoter"
     promoter        1112..1131
                     /created_by="lha16"
                     /label="T7 promoter"
     motif           1135..1147
                     /created_by="lha16"
                     /modified_by="lha16"
                     /label="Kozak"
     CDS             1144..1494
                     /created_by="joana"
                     /Color="#ffbb33"
                     /modified_by="lha16"
                     /label="PCV2 HUHe"
     CDS             1495..1566
                     /Color="#ff7400"
                     /created_by="lha16"
                     /label="GS-XTEN-GS linker"
     CDS             1564..4725
                     /created_by="User"
                     /modified_by="User"
                     /Color="#eeeeee"
                     /modified_by="lha16"
                     /label="nSaCas9(N580A)"
     CDS             3304..3306
                     /created_by="joana"
                     /modified_by="joana"
                     /modified_by="lha16"
                     /label="N580A"
     CDS             4726..4749
                     /Color="#ff7400"
                     /created_by="lha16"
                     /label="GS linker"
     CDS             4750..4803
                     /Color="#6bba70"
                     /created_by="lha16"
                     /label="BPNLS"
     CDS             4804..4827
                     /Color="#ff7400"
                     /created_by="lha16"
                     /label="GS linker"
     CDS             4828..6642
                     /created_by="joana"
                     /Color="#ee0077"
                     /modified_by="lha16"
                     /label="EcKlenow(-exo;D355A/D357A)"
     CDS             6643..6654
                     /created_by="lha16"
                     /Color="#ff7400"
                     /modified_by="lha16"
                     /label="GS linker"
     CDS             6655..6708
                     /created_by="lha16"
                     /Color="#6bba70"
                     /modified_by="lha16"
                     /label="BPNLS"
     CDS             6709..6711
                     /created_by="lha16"
                     /Color="#000000"
                     /modified_by="lha16"
                     /label
     terminator      6760..6987
                     /created_by="lha16"
                     /label="bGH_PA_terminator"
     terminator      7246..7377
                     /created_by="lha16"
                     /label="SV40_PA_terminator"
     rep_origin      complement(7445..8064)
                     /created_by="lha16"
                     /label="pBR322 origin"
     CDS             complement(8219..112)
                     /Color="#777777"
                     /created_by="lha16"
                     /label="Ampicillin"
ORIGIN      
        1 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
       61 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      241 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
      301 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      361 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      421 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      481 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      541 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      601 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      661 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      721 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      781 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      841 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      901 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      961 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     1021 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     1081 accgtcagat ccgctagaga tccgcggccg ctaatacgac tcactatagg gagagccgcc
     1141 accatgagcc catctaagaa aaacggcaga agcggacctc aacctcacaa gcggtgggtc
     1201 tttaccctga acaaccccag cgaagatgag agaaagaaga tccgggacct gcctatcagc
     1261 ctgttcgact acttcatcgt gggcgaggaa ggaaatgagg aaggccggac cccccacctg
     1321 cagggcttcg ccaacttcgt gaaaaagcag accttcaaca aggtgaagtg gtacctgggc
     1381 gctagatgcc acatcgagaa ggccaagggc accgaccagc agaacaaaga gtactgcagc
     1441 aaggaaggca atctgctgat ggaatgtggc gcccctagat cccagggaca gagaagcgga
     1501 ggctctagcg gctccgagac acccggcaca tctgagtccg ccacccctga gagcagcggc
     1561 ggcagcggga aacggaacta catcctgggg cttgacattg ggataaccag cgttggctac
     1621 ggaattattg attatgagac acgcgatgtg attgacgccg gggttaggct gttcaaagag
     1681 gccaacgttg aaaacaacga gggaagacgg agtaagcgcg gagcaagaag actcaagcgc
     1741 agacggagac atcggattca gagggtgaaa aagctgctct tcgattacaa tctcctgacc
     1801 gatcatagtg agctgagcgg aatcaacccc tacgaggcgc gagtgaaagg gctttcccag
     1861 aagctgtccg aagaggagtt ctccgccgcg ttgctgcacc tggccaaacg gaggggggtt
     1921 cacaatgtaa acgaagtgga ggaggacacg ggcaatgaac ttagtacgaa agaacagatc
     1981 agtaggaact ctaaggctct cgaagagaaa tacgtcgctg agttgcagct tgagagactg
     2041 aaaaaagacg gcgaagtacg cggatctatt aataggttca agacttcaga ttacgtaaag
     2101 gaagccaagc agctcctgaa agtacagaaa gcgtaccatc agctcgatca gagcttcatc
     2161 gatacctaca tagatttgct ggagacacgg aggacatact acgagggccc aggggaagga
     2221 tctccttttg ggtggaagga catcaaggaa tggtacgaga tgcttatggg acattgtaca
     2281 tattttccgg aggagctcag gagcgtcaag tacgcctaca atgccgacct gtacaatgcc
     2341 ctcaatgacc tcaataacct cgtgattacc agggacgaga acgagaagct ggagtactat
     2401 gaaaagttcc agattatcga gaatgtgttt aagcagaaga agaagccgac acttaagcag
     2461 attgcaaagg aaatcctcgt gaatgaggaa gatatcaagg gatacagagt gacaagtaca
     2521 ggcaagcccg agttcacaaa tctgaaggtg taccacgata ttaaggacat aaccgcacga
     2581 aaggagataa tcgaaaacgc tgagctcctc gatcagatcg caaaaattct taccatctac
     2641 cagtctagtg aggacattca ggaggaactg actaatctga acagtgagct cacccaagag
     2701 gaaattgagc agatttcaaa cctgaaaggc tacaccggga cgcacaatct gagcctcaaa
     2761 gcaatcaacc tcattctgga tgaactttgg cacacaaatg acaaccaaat tgccatattc
     2821 aaccgcctga aactggtgcc aaaaaaagtg gatctgtcac agcaaaagga aatccctaca
     2881 accttggttg acgattttat tctgtccccc gttgtcaagc ggagcttcat ccagtcaatc
     2941 aaggtgatca atgccatcat taaaaaatac ggattgccaa acgatataat tatcgagctt
     3001 gcacgagaga agaactcaaa ggacgcccag aagatgatta acgaaatgca gaagcgcaac
     3061 cgccagacaa acgaacgcat agaggaaatt ataagaacaa ccggcaaaga gaatgccaag
     3121 tatctgatcg agaaaatcaa gctgcacgac atgcaagaag gcaagtgcct gtactctctg
     3181 gaagctatcc cactcgaaga tctgctgaat aatccattca attacgaggt ggaccacatc
     3241 atccctagat ccgtaagctt tgacaattcc ttcaataaca aagttctggt taaacaggag
     3301 gaagcctcta aaaaagggaa ccggaccccg ttccagtacc tgagctccag tgacagcaag
     3361 attagctacg agacttttaa gaaacatatt ctgaatctgg ccaaaggcaa aggcaggatc
     3421 agcaagacca agaaggagta cctcctcgaa gaacgcgaca ttaacagatt tagtgtgcag
     3481 aaagatttca tcaaccgaaa ccttgtcgat actcggtacg ccacgagagg cctgatgaat
     3541 ctcctcagga gctacttccg cgtcaataat ctggacgtta aagtcaagag cataaatggg
     3601 ggattcacca gctttctgag gagaaagtgg aagtttaaga aggaacgaaa caaaggatac
     3661 aagcaccatg ctgaggatgc tttgatcatc gctaacgcgg actttatctt taaggaatgg
     3721 aaaaagctgg ataaggcaaa gaaagtgatg gaaaaccaga tgttcgagga gaagcaggca
     3781 gagtcaatgc ctgagatcga gacagagcag gaatacaagg aaattttcat cacccctcat
     3841 cagattaaac acataaagga cttcaaagac tataaatact ctcatagggt ggacaaaaaa
     3901 cccaatcgcg agctcattaa tgacaccctg tactcaacac ggaaggatga taaaggtaat
     3961 accttgattg tgaataatct taatggattg tatgacaaag ataacgacaa gctcaagaag
     4021 ctgatcaaca agtctccaga gaagctcctt atgtatcacc acgacccaca gacttatcag
     4081 aaattgaaac tgatcatgga gcaatacggg gatgagaaga acccactcta caaatattat
     4141 gaggaaacag gtaattacct gaccaagtac tccaagaagg ataacggacc agtgatcaaa
     4201 aagataaagt actatggcaa caaacttaat gcgcatttgg acataactga cgattacccc
     4261 aattctcgaa acaaggttgt gaagctctcc ctgaagcctt atagatttga cgtgtacctg
     4321 gataatgggg tttataaatt cgtcaccgtg aaaaatctgg acgtgatcaa aaaggagaac
     4381 tattatgaag taaactcaaa gtgctatgag gaggcgaaga agctgaagaa gatctccaat
     4441 caggccgagt tcatcgcttc cttctataat aacgatctca tcaagatcaa tggagagctt
     4501 tatcgcgtca ttggtgtgaa caatgacttg ctgaacagga tcgaagtcaa tatgatagac
     4561 attacctacc gggagtatct cgaaaacatg aatgataaac ggccgcctcg catcatcaag
     4621 acaatcgcat ctaaaactca gtcaataaaa aagtactcta ccgatatcct ggggaatctc
     4681 tatgaagtga agtcaaagaa gcacccacaa atcattaaaa aaggttccgg cggaagctct
     4741 ggtggcagca agcggaccgc cgacggctct gaattcgaga gccctaagaa gaaaagaaag
     4801 gtgagcggag gctctagcgg cggaagcgtt atctcctacg acaactacgt gaccatcctg
     4861 gacgaagaga cactgaaggc ttggatcgct aaactggaaa aagcccctgt gttcgccttc
     4921 gccaccgcca ccgactcctt agataacatc agcgccaacc tggtgggcct gtctttcgcc
     4981 atcgagcctg gcgtggccgc ctacatcccc gtggcccacg attacctgga tgccccagat
     5041 caaatcagca gagagagagc actggagctc ctgaagcccc tgctggagga cgagaaggcc
     5101 ctgaaggtgg gacagaacct gaaatacgat aggggaatcc tggctaatta cggcatcgag
     5161 ctgagaggca tcgccttcga taccatgctg gaatcttaca tcctgaacag cgtggccggc
     5221 cgccacgaca tggacagcct ggctgaacgg tggctgaagc acaaaaccat caccttcgaa
     5281 gagatcgccg gaaagggcaa gaaccagctg acatttaatc agatcgcact ggaagaagcc
     5341 ggcagatacg ccgctgagga cgccgacgtg acccttcagc tgcacctgaa gatgtggccc
     5401 gatctgcaga agcacaaggg cccactaaac gtgttcgaga acatcgagat gcctctggtg
     5461 cccgtcctga gcagaatcga gaggaatggc gtgaagatcg accccaaggt gctgcacaac
     5521 cacagcgagg aactgacact gcggctggcc gagctggaaa agaaggccca cgaaatcgcc
     5581 ggcgaggagt tcaacctgtc tagcaccaag cagctacaaa caatcctgtt cgagaagcag
     5641 ggcattaagc ctctgaaaaa gacacctggc ggagccccca gcaccagcga agaagtgctg
     5701 gaggaactgg cgctggatta ccctctgccc aaagtgatcc tggaatatag aggactggcc
     5761 aagctgaagt ctacatacac cgacaagctg cctctgatga tcaaccccaa gaccggccgg
     5821 gtgcatacat cttatcacca ggccgtgaca gccaccggcc ggctgagcag cacagatcct
     5881 aacctgcaaa acatcccagt gcggaacgaa gagggcagaa gaatcagaca ggctttcatt
     5941 gcccctgagg actacgtcat cgtgtctgct gactacagcc agatagaact gagaatcatg
     6001 gcccatctgt ctcgggacaa gggcctgctg accgcttttg cagaaggcaa ggacatccac
     6061 agagccaccg ctgccgaagt gttcggcctg cctctggaaa cagtgaccag cgagcaaaga
     6121 aggtccgcca aggccattaa cttcggcctg atctacggca tgagcgcctt tggcctggcc
     6181 agacagctca acatccctag aaaggaggcc cagaaataca tggacctgta cttcgagcgg
     6241 taccccggcg tgctggagta catggaacgg accagagccc aggctaaaga gcagggatac
     6301 gtggaaaccc tggatggccg gagactgtat ctgcctgaca tcaagagcag caacggcgcc
     6361 agaagagccg ccgctgagag agctgctatc aacgccccta tgcagggcac cgccgccgac
     6421 atcatcaagc gggccatgat cgccgtggac gcctggctgc aggccgagca gcctagagtg
     6481 cggatgatca tgcaggtgca cgacgagctg gtgtttgagg tgcacaagga cgacgtggat
     6541 gctgtggcca agcagattca ccagctgatg gaaaattgca ccagactgga cgtgccactg
     6601 ctggtcgagg ttggaagcgg agagaattgg gaccaggccc actctggcgg ctcaaaaaga
     6661 accgccgacg gcagcgaatt cgagtctccc aagaagaaga ggaaagtctg accggtcatc
     6721 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     6781 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     6841 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     6901 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     6961 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     7021 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     7081 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7141 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7201 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7261 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7321 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7381 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7441 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7501 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7561 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7621 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7681 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7741 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7801 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7861 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7921 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7981 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8041 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8101 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8161 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8221 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8281 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8341 gtgctgcaat gataccgcgc gaaccacgct caccggctcc agatttatca gcaataaacc
     8401 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8461 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8521 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8581 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8641 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8701 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8761 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8821 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8881 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8941 gttcgatgta acccactcgt gcaccca
//
