LOCUS       pSCL72                 10407 bp    DNA     circular UNA 19-JAN-2022
DEFINITION  Gibson/Homology Cloning ligation product.
ACCESSION   urn.local...1p7-d5b6sg9
VERSION     urn.local...1p7-d5b6sg9
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"
     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"
     linker          4865..4924
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="linker"
     CDS             <4925..>5908
                     /standard_name="RT Ec86 Sc optimized"
     tag             5882..5905
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="SV40 NLS"
     misc_feature    5920..6166
                     /standard_name="CYC1 terminator"
     protein_bind    complement(6256..6276)
                     /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"
     protein_bind    6258..6274
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="lac operator"
     promoter        complement(6282..6312)
                     /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(6618..7300)
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="ColE1"
     CDS             complement(7395..8255)
                     /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(8256..8360)
                     /gene="bla"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="AmpR promoter"
     promoter        8772..8958
                     /gene="S. cerevisiae HIS3"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="HIS3 promoter"
     CDS             8959..9618
                     /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              9619..9819
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="dubious DED1 orf"
     rep_origin      9878..10333
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 agacggcggt acgtcttccg gtggctctgg tcccgctggt agctctggcg caacgagcgg
     4921 tggtaagtcc gctgaatact tgaacacctt cagattgaga aatttgggtt tgccagtcat
     4981 gaacaacttg cacgatatgt ctaaggctac cagaatttct gttgaaacct tgagattatt
     5041 gatctacacc gccgatttca gatacagaat ctacactgtt gaaaagaagg gtccagaaaa
     5101 gagaatgaga actatctacc aaccatccag agaattgaag gctttacaag gttgggtttt
     5161 gagaaacatc ttggacaagt tatcctcctc cccattttct atcggttttg aaaagcacca
     5221 atccatcttg aacaatgcca ctccacatat tggtgccaac ttcattttga acatcgactt
     5281 ggaagatttc ttcccatctt tgactgctaa caaggttttc ggtgttttcc attctttggg
     5341 ttacaacaga ttgatctcct ctgttttgac caagatttgc tgctacaaga acttgttgcc
     5401 acaaggtgct ccatcttctc caaaattggc taacttgatt tgctccaagt tggactacag
     5461 aattcaaggt tatgctggtt ctagaggttt gatatacact agatacgctg atgacttgac
     5521 cttgtctgct caatctatga agaaagttgt taaggccaga gacttcttgt tctccattat
     5581 tccatcagaa ggtttggtca tcaactccaa aaagacctgt atttctggtc caagatccca
     5641 aagaaaagtt accggtttgg ttatctccca agaaaaggtt ggtattggta gagaaaagta
     5701 caaagaaatc agagccaaga tccaccatat tttctgcggt aaatcctccg aaatcgaaca
     5761 tgttagaggt tggttgtctt tcatcttgtc cgttgattct aagtcccaca gaagattgat
     5821 tacctacatc tctaagttgg aaaagaagta cggtaagaac ccattgaaca aggctaagac
     5881 tccaccaaag aagaaaagaa aggtctgatc tcttctcgag tcatgtaatt agttatgtca
     5941 cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa
     6001 cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt
     6061 atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg
     6121 aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttcacc gcggtggagc
     6181 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     6241 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc
     6301 ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc
     6361 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6421 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6481 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6541 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7021 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7261 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7321 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7381 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7441 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7501 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7561 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7621 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7681 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7741 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7801 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7861 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7921 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7981 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8041 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8101 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8161 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8221 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8281 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8341 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     8401 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     8461 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     8521 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     8581 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     8641 aaattcccgt tttaagagct tggtgagcgc taggagtcac tgccaggtat cgtttgaaca
     8701 cggcattagt cagggaagtc ataacacagt cctttcccgc aattttcttt ttctattact
     8761 cttggcctcc tctagtacac tctatatttt tttatgcctc ggtaatgatt ttcatttttt
     8821 tttttcccct agcggatgac tctttttttt tcttagcgat tggcattatc acataatgaa
     8881 ttatacatta tataaagtaa tgtgatttct tcgaagaata tactaaaaaa tgagcaggca
     8941 agataaacga aggcaaagat gacagagcag aaagccctag taaagcgtat tacaaatgaa
     9001 accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga gcactcgatc
     9061 ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca agtgattaac
     9121 gtccacacag gtatagggtt tctggaccat atgatacatg ctctggccaa gcattccggc
     9181 tggtcgctaa tcgttgagtg cattggtgac ttacacatag acgaccatca caccactgaa
     9241 gactgcggga ttgctctcgg tcaagctttt aaagaggccc tactggcgcg tggagtaaaa
     9301 aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt ggtagatctt
     9361 tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc
     9421 tcttgcgaga tgatcccgca ttttcttgaa agctttgcag aggctagcag aattaccctc
     9481 cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc gttcaaggct
     9541 cttgcggttg ccataagaga agccacctcg cccaatggta ccaacgatgt tccctccacc
     9601 aaaggtgttc ttatgtagtg acaccgatta tttaaagctg cagcatacga tatatataca
     9661 tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt atgatactga
     9721 agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc gctttccttt
     9781 tttctttttg ctttttcttt ttttttctct tgaactcgac ggatctatgc ggtgtgaaat
     9841 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaaacgt taatattttg
     9901 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     9961 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
    10021 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
    10081 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
    10141 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
    10201 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
    10261 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
    10321 ctacagggcg cgtcgcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
    10381 gtgcgggcct cttcgctatt acgccag
//
