LOCUS       pCAG-hSpCas9-NLS-FLAG-P2A-EGFP_(MSP2582)        9829 bp    DNA     circular UNA 02-OCT-2024
DEFINITION  .
ACCESSION   urn.local...7j-ia1c4os
VERSION     urn.local...7j-ia1c4os
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     regulatory      18..1718
                     /note="Geneious type: promoter"
                     /standard_name="CAG promoter"
     misc_feature    1763..1772
                     /note="Geneious type: motif"
                     /standard_name="Kozak"
     CDS             1769..5872
                     /standard_name="hSpCas9"
     CDS             5981..6046
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIK
                     KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE
                     ESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAH
                     MIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSK
                     SRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLD
                     NLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTL
                     LKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVK
                     LNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPY
                     YVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL
                     PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKE
                     DYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLT
                     LFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDF
                     LKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT
                     VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE
                     HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKV
                     LTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA
                     GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIG
                     KATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS
                     MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVV
                     AKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLF
                     ELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQH
                     KHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA
                     PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDKRPAATKKAG
                     QAKKKKDYKDDDDKGSGATNFSLLKQAGDVEENPGPTEYKPTVRLATRDDVPRAVRTL
                     AAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPES
                     VEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGL
                     GSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKP
                     GA*"
                     /standard_name="P2A"
     CDS             6047..6763
                     /standard_name="EGFP"
     regulatory      6773..7297
                     /note="Geneious type: Terminator"
                     /standard_name="rb_glob_PA term"
     regulatory      7504..7706
                     /note="Geneious type: Promoter"
                     /standard_name="SV40 prom"
     misc_feature    7551..7628
                     /note="Geneious type: Origin of Replication"
                     /standard_name="SV40 origin"
     misc_feature    complement(8064..8683)
                     /note="Geneious type: Origin of Replication"
                     /standard_name="pBR322 origin"
     misc_feature    complement(8838..9698)
                     /note="Geneious type: Marker"
                     /standard_name="amp marker"
     regulatory      complement(9740..9768)
                     /note="Geneious type: Promoter"
                     /standard_name="amp prom"
ORIGIN      
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
     1741 gcgggcccgg gatccaccgg tcgccaccat ggataaaaag tattctattg gtttagacat
     1801 cggcactaat tccgttggat gggctgtcat aaccgatgaa tacaaagtac cttcaaagaa
     1861 atttaaggtg ttggggaaca cagaccgtca ttcgattaaa aagaatctta tcggtgccct
     1921 cctattcgat agtggcgaaa cggcagaggc gactcgcctg aaacgaaccg ctcggagaag
     1981 gtatacacgt cgcaagaacc gaatatgtta cttacaagaa atttttagca atgagatggc
     2041 caaagttgac gattctttct ttcaccgttt ggaagagtcc ttccttgtcg aagaggacaa
     2101 gaaacatgaa cggcacccca tctttggaaa catagtagat gaggtggcat atcatgaaaa
     2161 gtacccaacg atttatcacc tcagaaaaaa gctagttgac tcaactgata aagcggacct
     2221 gaggttaatc tacttggctc ttgcccatat gataaagttc cgtgggcact ttctcattga
     2281 gggtgatcta aatccggaca actcggatgt cgacaaactg ttcatccagt tagtacaaac
     2341 ctataatcag ttgtttgaag agaaccctat aaatgcaagt ggcgtggatg cgaaggctat
     2401 tcttagcgcc cgcctctcta aatcccgacg gctagaaaac ctgatcgcac aattacccgg
     2461 agagaagaaa aatgggttgt tcggtaacct tatagcgctc tcactaggcc tgacaccaaa
     2521 ttttaagtcg aacttcgact tagctgaaga tgccaaattg cagcttagta aggacacgta
     2581 cgatgacgat ctcgacaatc tactggcaca aattggagat cagtatgcgg acttattttt
     2641 ggctgccaaa aaccttagcg atgcaatcct cctatctgac atactgagag ttaatactga
     2701 gattaccaag gcgccgttat ccgcttcaat gatcaaaagg tacgatgaac atcaccaaga
     2761 cttgacactt ctcaaggccc tagtccgtca gcaactgcct gagaaatata aggaaatatt
     2821 ctttgatcag tcgaaaaacg ggtacgcagg ttatattgac ggcggagcga gtcaagagga
     2881 attctacaag tttatcaaac ccatattaga gaagatggat gggacggaag agttgcttgt
     2941 aaaactcaat cgcgaagatc tactgcgaaa gcagcggact ttcgacaacg gtagcattcc
     3001 acatcaaatc cacttaggcg aattgcatgc tatacttaga aggcaggagg atttttatcc
     3061 gttcctcaaa gacaatcgtg aaaagattga gaaaatccta acctttcgca taccttacta
     3121 tgtgggaccc ctggcccgag ggaactctcg gttcgcatgg atgacaagaa agtccgaaga
     3181 aacgattact ccatggaatt ttgaggaagt tgtcgataaa ggtgcgtcag ctcaatcgtt
     3241 catcgagagg atgaccaact ttgacaagaa tttaccgaac gaaaaagtat tgcctaagca
     3301 cagtttactt tacgagtatt tcacagtgta caatgaactc acgaaagtta agtatgtcac
     3361 tgagggcatg cgtaaacccg cctttctaag cggagaacag aagaaagcaa tagtagatct
     3421 gttattcaag accaaccgca aagtgacagt taagcaattg aaagaggact actttaagaa
     3481 aattgaatgc ttcgattctg tcgagatctc cggggtagaa gatcgattta atgcgtcact
     3541 tggtacgtat catgacctcc taaagataat taaagataag gacttcctgg ataacgaaga
     3601 gaatgaagat atcttagaag atatagtgtt gactcttacc ctctttgaag atcgggaaat
     3661 gattgaggaa agactaaaaa catacgctca cctgttcgac gataaggtta tgaaacagtt
     3721 aaagaggcgt cgctatacgg gctggggacg attgtcgcgg aaacttatca acgggataag
     3781 agacaagcaa agtggtaaaa ctattctcga ttttctaaag agcgacggct tcgccaatag
     3841 gaactttatg cagctgatcc atgatgactc tttaaccttc aaagaggata tacaaaaggc
     3901 acaggtttcc ggacaagggg actcattgca cgaacatatt gcgaatcttg ctggttcgcc
     3961 agccatcaaa aagggcatac tccagacagt caaagtagtg gatgagctag ttaaggtcat
     4021 gggacgtcac aaaccggaaa acattgtaat cgagatggca cgcgaaaatc aaacgactca
     4081 gaaggggcaa aaaaacagtc gagagcggat gaagagaata gaagagggta ttaaagaact
     4141 gggcagccag atcttaaagg agcatcctgt ggaaaatacc caattgcaga acgagaaact
     4201 ttacctctat tacctacaaa atggaaggga catgtatgtt gatcaggaac tggacataaa
     4261 ccgtttatct gattacgacg tcgatcacat tgtaccccaa tcctttttga aggacgattc
     4321 aatcgacaat aaagtgctta cacgctcgga taagaaccga gggaaaagtg acaatgttcc
     4381 aagcgaggaa gtcgtaaaga aaatgaagaa ctattggcgg cagctcctaa atgcgaaact
     4441 gataacgcaa agaaagttcg ataacttaac taaagctgag aggggtggct tgtctgaact
     4501 tgacaaggcc ggatttatta aacgtcagct cgtggaaacc cgccaaatca caaagcatgt
     4561 tgcacagata ctagattccc gaatgaatac gaaatacgac gagaacgata agctgattcg
     4621 ggaagtcaaa gtaatcactt taaagtcaaa attggtgtcg gacttcagaa aggattttca
     4681 attctataaa gttagggaga taaataacta ccaccatgcg cacgacgctt atcttaatgc
     4741 cgtcgtaggg accgcactca ttaagaaata cccgaagcta gaaagtgagt ttgtgtatgg
     4801 tgattacaaa gtttatgacg tccgtaagat gatcgcgaaa agcgaacagg agataggcaa
     4861 ggctacagcc aaatacttct tttattctaa cattatgaat ttctttaaga cggaaatcac
     4921 tctggcaaac ggagagatac gcaaacgacc tttaattgaa accaatgggg agacaggtga
     4981 aatcgtatgg gataagggcc gggacttcgc gacggtgaga aaagttttgt ccatgcccca
     5041 agtcaacata gtaaagaaaa ctgaggtgca gaccggaggg ttttcaaagg aatcgattct
     5101 tccaaaaagg aatagtgata agctcatcgc tcgtaaaaag gactgggacc cgaaaaagta
     5161 cggtggcttc gatagcccta cagttgccta ttctgtccta gtagtggcaa aagttgagaa
     5221 gggaaaatcc aagaaactga agtcagtcaa agaattattg gggataacga ttatggagcg
     5281 ctcgtctttt gaaaagaacc ccatcgactt ccttgaggcg aaaggttaca aggaagtaaa
     5341 aaaggatctc ataattaaac taccaaagta tagtctgttt gagttagaaa atggccgaaa
     5401 acggatgttg gctagcgccg gagagcttca aaaggggaac gaactcgcac taccgtctaa
     5461 atacgtgaat ttcctgtatt tagcgtccca ttacgagaag ttgaaaggtt cacctgaaga
     5521 taacgaacag aagcaacttt ttgttgagca gcacaaacat tatctcgacg aaatcataga
     5581 gcaaatttcg gaattcagta agagagtcat cctagctgat gccaatctgg acaaagtatt
     5641 aagcgcatac aacaagcaca gggataaacc catacgtgag caggcggaaa atattatcca
     5701 tttgtttact cttaccaacc tcggcgctcc agccgcattc aagtattttg acacaacgat
     5761 agatcgcaaa cgatacactt ctaccaagga ggtgctagac gcgacactga ttcaccaatc
     5821 catcacggga ttatatgaaa ctcggataga tttgtcacag cttgggggtg acggatcccc
     5881 caagaagaag aggaaagtct cgagcgacta caaagaccat gacggtgatt ataaagatca
     5941 tgacatcgat tacaaggatg acgatgacaa gggaggcggt ggaagcggcg caacaaactt
     6001 ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgatgg tgagcaaggg
     6061 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     6121 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     6181 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     6241 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     6301 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     6361 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     6421 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     6481 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     6541 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     6601 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     6661 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     6721 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtgaagcg gccgcactcc
     6781 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     6841 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     6901 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     6961 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     7021 gtatttggtt tagagtttgg caacatatgc catatgctgg ctgccatgaa caaaggtggc
     7081 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     7141 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     7201 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     7261 tcccagtcat agctgtccct cttctcttat gaagatccct cgacctgcag cccaagcttg
     7321 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7381 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7441 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     7501 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     7561 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     7621 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7681 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     7741 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7801 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     7861 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7921 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7981 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     8041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     8101 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     8161 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     8221 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     8281 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8341 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8401 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8461 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     8521 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8581 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8701 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8761 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8821 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8881 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8941 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     9001 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     9061 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     9121 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9181 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9241 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9301 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9361 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9421 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9481 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9541 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9601 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9661 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9721 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9781 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
