LOCUS       pSCL103                10404 bp    DNA     circular UNA 19-JAN-2022
DEFINITION  cas9 p2a eco1RT.
ACCESSION   urn.local...136-dc0q8g9
VERSION     urn.local...136-dc0q8g9
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     tag             complement(12..35)
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="SV40 NLS"
     promoter        39..271
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="Gal10p"
     promoter        272..702
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="Gal1/10 promoter"
     misc_feature    complement(<312..>323)
                     /note="asp-tag (polyaspartate) used for protein
                     immobilization and purification"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on protein similarity"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on protein similarity"
                     /standard_name="polyaspartate"
     nls             731..763
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="SV40"
     CDS             <764..>4864
                     /codon_start=1
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="Cas9"
     p2a             4865..4921
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="P2A"
     CDS             <4922..>5878
                     /standard_name="RT Ec86 Sc optimized"
     tag             5879..5902
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="SV40 NLS"
     misc_feature    5917..6163
                     /standard_name="CYC1 terminator"
     protein_bind    complement(6253..6273)
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="lacO"
     promoter        complement(6279..6309)
                     /note="promoter for the E. coli lac operon"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="lac promoter"
     misc_feature    complement(6615..7297)
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="ColE1"
     CDS             complement(7392..8252)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="AmpR"
     promoter        complement(8253..8357)
                     /gene="bla"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="AmpR promoter"
     promoter        8769..8955
                     /gene="S. cerevisiae HIS3"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="HIS3 promoter"
     CDS             8956..9615
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase,
                     required for histidine biosynthesis"
                     /note="yeast auxotrophic marker"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="HIS3"
     nc              9616..9816
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="dubious DED1 orf"
     rep_origin      9875..10330
ORIGIN      
        1 tcagcggact tgacctttct tttcttcttt ggtggcattt atattgaatt ttcaaaaatt
       61 cttacttttt ttttggatgg acgcaaagaa gtttaataat catattacat ggcattacca
      121 ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa gagccccatt
      181 atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa ctgctcattg
      241 ctatattgaa gtacggatta gaagccgccg agcgggcgac agccctccga cggaagactc
      301 tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc
      361 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
      421 attggcagta acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg
      481 atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
      541 tttttgatct attaacagat atataaatgg aaaagctgca taaccacttt aactaatact
      601 ttcaacattt tcagtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa
      661 ttgttaatat acctctatac tttaacgtca aggagaaaaa acaactagtg gatcccccgg
      721 gaaaaaaatg agcagggctg accccaagaa gaagaggaag gtggacaaga agtactccat
      781 tgggctcgat atcggcacaa acagcgtcgg ctgggccgtc attacggacg agtacaaggt
      841 gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct
      901 cattggcgcc ctcctgttcg actccgggga gacggccgaa gccacgcggc tcaaaagaac
      961 agcacggcgc agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag
     1021 taatgagatg gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt
     1081 ggaggaggat aaaaagcacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc
     1141 gtaccatgaa aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga
     1201 taaggctgac ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca
     1261 cttcctcatc gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca
     1321 actggttcag acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga
     1381 cgccaaagca atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc
     1441 acagctccct ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg
     1501 gctgaccccc aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag
     1561 caaagacacc tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc
     1621 agaccttttt ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg
     1681 agtgaacacg gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga
     1741 gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta
     1801 caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc
     1861 aagccaggag gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga
     1921 ggagctgctg gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa
     1981 tggaagcatc ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga
     2041 ggatttctac ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg
     2101 gataccctac tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg
     2161 caaatcagaa gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc
     2221 tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt
     2281 gcttcctaaa cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt
     2341 caaatacgtc acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc
     2401 tatcgtggac ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga
     2461 ctatttcaaa aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt
     2521 caacgcatcc ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct
     2581 ggacaatgag gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga
     2641 agatagggag atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt
     2701 catgaaacag ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat
     2761 caatgggatc cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg
     2821 atttgccaac cggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga
     2881 catccagaaa gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct
     2941 tgcaggtagc ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact
     3001 cgtcaaagta atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa
     3061 ccaaactacc cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg
     3121 tataaaagaa ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca
     3181 gaatgagaag ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga
     3241 actggacatc aatcggctct ccgactacga cgtggatcat atcgtgcccc agtcttttct
     3301 caaagatgat tctattgata ataaagtgtt gacaagatcc gataaaaata gagggaagag
     3361 tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct
     3421 gaacgccaaa ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg
     3481 cctgtctgag ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat
     3541 caccaagcac gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga
     3601 caaactgatt cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag
     3661 aaaggacttt cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc
     3721 ctacctgaat gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga
     3781 atttgtttac ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca
     3841 ggaaataggc aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa
     3901 gaccgagatt acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg
     3961 agaaacagga gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct
     4021 gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa
     4081 ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga
     4141 ccccaagaaa tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc
     4201 caaagtggag aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac
     4261 aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata
     4321 taaagaggtc aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga
     4381 aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc
     4441 actgccctct aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg
     4501 gtctcccgaa gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga
     4561 tgagatcatc gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct
     4621 cgataaggtg ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga
     4681 aaacattatc cacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt
     4741 cgacaccacc atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact
     4801 gattcatcag tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg
     4861 agacgcaaca aacttctctc tgctgaaaca agccggagat gtcgaagaga atcctggacc
     4921 gaagtccgct gaatacttga acaccttcag attgagaaat ttgggtttgc cagtcatgaa
     4981 caacttgcac gatatgtcta aggctaccag aatttctgtt gaaaccttga gattattgat
     5041 ctacaccgcc gatttcagat acagaatcta cactgttgaa aagaagggtc cagaaaagag
     5101 aatgagaact atctaccaac catccagaga attgaaggct ttacaaggtt gggttttgag
     5161 aaacatcttg gacaagttat cctcctcccc attttctatc ggttttgaaa agcaccaatc
     5221 catcttgaac aatgccactc cacatattgg tgccaacttc attttgaaca tcgacttgga
     5281 agatttcttc ccatctttga ctgctaacaa ggttttcggt gttttccatt ctttgggtta
     5341 caacagattg atctcctctg ttttgaccaa gatttgctgc tacaagaact tgttgccaca
     5401 aggtgctcca tcttctccaa aattggctaa cttgatttgc tccaagttgg actacagaat
     5461 tcaaggttat gctggttcta gaggtttgat atacactaga tacgctgatg acttgacctt
     5521 gtctgctcaa tctatgaaga aagttgttaa ggccagagac ttcttgttct ccattattcc
     5581 atcagaaggt ttggtcatca actccaaaaa gacctgtatt tctggtccaa gatcccaaag
     5641 aaaagttacc ggtttggtta tctcccaaga aaaggttggt attggtagag aaaagtacaa
     5701 agaaatcaga gccaagatcc accatatttt ctgcggtaaa tcctccgaaa tcgaacatgt
     5761 tagaggttgg ttgtctttca tcttgtccgt tgattctaag tcccacagaa gattgattac
     5821 ctacatctct aagttggaaa agaagtacgg taagaaccca ttgaacaagg ctaagactcc
     5881 accaaagaag aaaagaaagg tctgatctct tctcgagtca tgtaattagt tatgtcacgc
     5941 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     6001 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     6061 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     6121 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttcaccgcg gtggagctcc
     6181 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     6241 tttcctgtgt gaaattgtta tccgctcaca attccacaca acataggagc cggaagcata
     6301 aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca
     6361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6421 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6481 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6541 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6601 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6661 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6721 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6781 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6841 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6901 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6961 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7021 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7081 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7141 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7201 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7261 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7321 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7381 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7441 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7501 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7561 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7621 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7681 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7741 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7801 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7861 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7921 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7981 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8041 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8101 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8161 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8221 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8281 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8341 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     8401 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     8461 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     8521 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     8581 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa
     8641 ttcccgtttt aagagcttgg tgagcgctag gagtcactgc caggtatcgt ttgaacacgg
     8701 cattagtcag ggaagtcata acacagtcct ttcccgcaat tttctttttc tattactctt
     8761 ggcctcctct agtacactct atattttttt atgcctcggt aatgattttc attttttttt
     8821 ttcccctagc ggatgactct ttttttttct tagcgattgg cattatcaca taatgaatta
     8881 tacattatat aaagtaatgt gatttcttcg aagaatatac taaaaaatga gcaggcaaga
     8941 taaacgaagg caaagatgac agagcagaaa gccctagtaa agcgtattac aaatgaaacc
     9001 aagattcaga ttgcgatctc tttaaagggt ggtcccctag cgatagagca ctcgatcttc
     9061 ccagaaaaag aggcagaagc agtagcagaa caggccacac aatcgcaagt gattaacgtc
     9121 cacacaggta tagggtttct ggaccatatg atacatgctc tggccaagca ttccggctgg
     9181 tcgctaatcg ttgagtgcat tggtgactta cacatagacg accatcacac cactgaagac
     9241 tgcgggattg ctctcggtca agcttttaaa gaggccctac tggcgcgtgg agtaaaaagg
     9301 tttggatcag gatttgcgcc tttggatgag gcactttcca gagcggtggt agatctttcg
     9361 aacaggccgt acgcagttgt cgaacttggt ttgcaaaggg agaaagtagg agatctctct
     9421 tgcgagatga tcccgcattt tcttgaaagc tttgcagagg ctagcagaat taccctccac
     9481 gttgattgtc tgcgaggcaa gaatgatcat caccgtagtg agagtgcgtt caaggctctt
     9541 gcggttgcca taagagaagc cacctcgccc aatggtacca acgatgttcc ctccaccaaa
     9601 ggtgttctta tgtagtgaca ccgattattt aaagctgcag catacgatat atatacatgt
     9661 gtatatatgt atacctatga atgtcagtaa gtatgtatac gaacagtatg atactgaaga
     9721 tgacaaggta atgcatcatt ctatacgtgt cattctgaac gaggcgcgct ttcctttttt
     9781 ctttttgctt tttctttttt tttctcttga actcgacgga tctatgcggt gtgaaatacc
     9841 gcacagatgc gtaaggagaa aataccgcat caggaaattg taaacgttaa tattttgtta
     9901 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     9961 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
    10021 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
    10081 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
    10141 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
    10201 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
    10261 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
    10321 cagggcgcgt cgcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
    10381 cgggcctctt cgctattacg ccag
//
