LOCUS       .                       5809 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     protein_bind    18..51
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            106..1462
                     /label="kanMX"
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        1681..1949
                     /label="SNR52 promoter"
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        1970..2044
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     terminator      2049..2068
                     /label="SUP4 terminator"
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     rep_origin      2114..3274
                     /label="2μ ori"
                     /note="yeast 2μ plasmid origin of replication"
     terminator      complement(3320..3339)
                     /label="SUP4 terminator"
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     misc_RNA        complement(3344..3418)
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        complement(3439..3707)
                     /label="SNR52 promoter"
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     rep_origin      complement(4011..4599)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4773..5630)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(5631..5735)
                     /label="AmpR promoter"
ORIGIN
        1 ggtcgacaac ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtctag
       61 agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag
      121 aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg
      181 tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc
      241 acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc
      301 ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag
      361 gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt
      421 tctcacatca catccgaaca taaacaacca tgggtaagga aaagactcac gtttcgaggc
      481 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
      541 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
      601 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
      661 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
      721 atgcatggtt actcaccact gcgatccccg gcaaaacagc attccaggta ttagaagaat
      781 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
      841 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
      901 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
      961 ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag
     1021 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     1081 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     1141 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     1201 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     1261 cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta
     1321 tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga
     1381 catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat
     1441 gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtgtcgaa
     1501 aacgagctct cgagaaccct taatataact tcgtataatg tatgctatac gaagttatta
     1561 ggtgatatca gatccactag tggcctatgc cacctttcga gaggacgatg cccgtgtcta
     1621 aatgattcga ccagcctaag aatgttcaac ccctcactaa agggaacaaa agctggagct
     1681 tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt
     1741 ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt
     1801 agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt
     1861 caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga
     1921 aacttctccg cagtgaaaga taaatgatca cttgaagatt ctttagtgtg ttttagagct
     1981 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     2041 ggtggtgctt tttttgtttt ttatgtcttc gagtcatgta attagttatg tcacgcttac
     2101 gttcacgccc tccacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat
     2161 atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag
     2221 ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa
     2281 gttcctattc tgtagaaagt ataggaacct cagagcgctt ttgaaaacca aaagcgctct
     2341 gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt aacgagctac taaaatattg
     2401 cgaataccgc ttccacaaac attgctcaaa agtatctctt tgctatatat ctctgtgcta
     2461 tatccctata taacctaccc atccaccttt cgctccttga acttgcatct aaactcgacc
     2521 tctacatttt ttatgtttat ctctagtatt actctttaga caaaaaaatt gtagtaagaa
     2581 ctattcatag agtgaatcga aaacaatacg aaaatgtaaa catttcctat acgtagtata
     2641 tagagacaaa atagaagaaa ccgttcataa ttttctgacc aatgaagaat catcaacgct
     2701 atcactttct gttcacaaag tatgcgcaat ccacatcggt atagaatata atcggggatg
     2761 cctttatctt gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg
     2821 agtcaggctt tttttatgga agagaaaata gacaccaaag tagccttctt ctaaccttaa
     2881 cggacctaca gtgcaaaaag ttatcaagag actgcattat agagcgcaca aaggagaaaa
     2941 aaagtaatct aagatgcttt gttagaaaaa tagcgctctc gggatgcatt tttgtagaac
     3001 aaaaaagaag tatagattct ttgttggtaa aatagcgctc tcgcgttgca tttctgttct
     3061 gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt
     3121 tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct
     3181 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
     3241 tttgttctac aaaatgaagc acagatgctt cgttggaggg cgtgaacgta agcgtgacat
     3301 aactaattac atgactcgaa gacataaaaa acaaaaaaag caccaccgac tcggtgccac
     3361 tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaactc
     3421 ctccatagag aacgtatcga tcatttatct ttcactgcgg agaagtttcg aacgccgaaa
     3481 catgcgcacc aactttcact tctacagcgt ttgaccaaaa tcttttgaac agaacattgt
     3541 agggtgtgaa aaaatgcgca cctttaccgc tagcccaaga gggcactaca aaatctagag
     3601 ttgtacttca aacgtacatg taatcacctt gtatatactc gaaagaaaac atcaagtttc
     3661 tgtataaata tgagtgaaag cataatcata cattatcttt tcaaagaagc tccagctttt
     3721 gttcccttta gtgagggtat tcacgtagac ggataggtat agccagacat cagcagcata
     3781 cttcgggaac cgtaggctct agagagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3841 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3901 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3961 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4021 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4081 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4141 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4201 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4261 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4321 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4381 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4441 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4501 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4561 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4621 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4681 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4741 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4801 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4861 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4921 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4981 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5041 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5101 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5161 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5221 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5281 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5341 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5401 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5461 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5521 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5581 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5641 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5701 tgtatttaga aaaataaaca aataggggtt ccgcgcactc gagactatat gtgaaggcat
     5761 ggctatggca cggcagacat tccgccagat catcaatagg cacgcggcc
//
