LOCUS       .                       6103 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      229..1211
                     /label="2μ ori"
                     /note="yeast 2μ plasmid origin of replication"
     promoter        1238..1342
                     /label="AmpR promoter"
     CDS             1343..2200
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      2374..2962
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    3250..3271
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3286..3316
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3324..3340
                     /label="lac operator"
                     /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-β-D-thiogalactopyranoside (IPTG)."
     primer_bind     3348..3364
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3385..3403
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3421..3689
                     /label="SNR52 promoter"
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        3710..3784
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     terminator      3789..3808
                     /label="SUP4 terminator"
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      3814..4061
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     promoter        complement(4080..4098)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4108..4124)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4597..4630)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 atacgaagtt atattaaggg ttgtcgacct gcagcgtacg aaggtgccta ttgatgatct
       61 ggcggaatgt ctgccgtgcc atagccatgc cttcacatat agtacaggca acacgcagat
      121 ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc gcgttgcatt ttcggaagcg
      181 ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt cctattctct agaaagtata
      241 ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa gacgcacttt caaaaaacca
      301 aaaacgcacc ggactgtaac gagctactaa aatattgcga ataccgcttc cacaaacatt
      361 gctcaaaagt atctctttgc tatatatctc tgtgctatat ccctatataa cctacccatc
      421 cacctttcgc tccttgaact tgcatctaaa ctcgacctct acatttttta tgtttatctc
      481 tagtattact ctttagacaa aaaaattgta gtaagaacta ttcatagagt gaatcgaaaa
      541 caatacgaaa atgtaaacat ttcctatacg tagtatatag agacaaaata gaagaaaccg
      601 ttcataattt tctgaccaat gaagaatcat caacgctatc actttctgtt cacaaagtat
      661 gcgcaatcca catcggtata gaatataatc ggggatgcct ttatcttgaa aaaatgcacc
      721 cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt caggcttttt ttatggaaga
      781 gaaaatagac accaaagtag ccttcttcta accttaacgg acctacagtg caaaaagtta
      841 tcaagagact gcattataga gcgcacaaag gagaaaaaaa gtaatctaag atgctttgtt
      901 agaaaaatag cgctctcggg atgcattttt gtagaacaaa aaagaagtat agattctttg
      961 ttggtaaaat agcgctctcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt
     1021 tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt acaaaaatga agcacagatt
     1081 cttcgttggt aaaatagcgc tttcgcgttg catttctgtt ctgtaaaaat gcagctcaga
     1141 ttctttgttt gaaaaattag cgctctcgcg ttgcattttt gttctacaaa atgaagcaca
     1201 gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1261 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1321 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1381 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1441 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1501 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1561 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1621 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1681 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1741 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1801 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1861 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1921 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1981 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2041 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2101 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2161 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2221 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2281 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2521 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2821 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2881 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2941 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3001 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3061 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3121 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3181 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3241 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     3301 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     3361 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     3421 tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt
     3481 ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt
     3541 agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt
     3601 caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga
     3661 aacttctccg cagtgaaaga taaatgatcg atacgttctc tatggaggag ttttagagct
     3721 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     3781 ggtggtgctt tttttgtttt ttatgtcttc gagtcatgta attagttatg tcacgcttac
     3841 gttcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     3901 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     3961 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     4021 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc
     4081 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     4141 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4201 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4261 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4321 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4381 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4441 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4501 gggccgttga acattcttag gctggtcgaa tcatttagac acgggcatcg tcctctcgaa
     4561 aggtggctag gccactagtg gatctgatat cacctaataa cttcgtatag catacattat
     4621 acgaagttat attaagggtt ctcgagagct cgttttattt aggttctatc gaggagaaaa
     4681 agcgacaaga agagatagac catggataaa tgattatgtt ctaaacactc ctcagaagct
     4741 catcgaactg tcatcctgcg tgaagattaa aatccaactt agaaatttcg agcttacgga
     4801 gacaatcata tgggagaagc aattggaaga tagaaaaaag gtactcggta cataaatata
     4861 tgtgattctg ggtagaagat cggtctgcat tggatggtgg taacgcattt ttttacacac
     4921 attacttgcc tcgagcatca aatggtggtt attcgtggat ctatatcacg tgatttgctt
     4981 aagaattgtc gttcatggtg acacttttag ctttgacatg attaagctca tctcaattga
     5041 tgttatctaa agtcatttca actatctaag atgtggttgt gattgggcca ttttgtgaaa
     5101 gccagtacgc cagcgtcaat acactcccgt caattagttg caccatgtcc acaaaatcat
     5161 ataccagtag agctgagact catgcaagtc cggttgcatc gaaactttta cgtttaatgg
     5221 atgaaaagaa gaccaatttg tgtgcttctc ttgacgttcg ttcgactgat gagctattga
     5281 aacttgttga aacgttgggt ccatacattt gccttttgaa aacacacgtt gatatcttgg
     5341 atgatttcag ttatgagggt actgtcgttc cattgaaagc attggcagag aaatacaagt
     5401 tcttgatatt tgaggacaga aaattcgccg atatcggtaa cacagtcaaa ttacaatata
     5461 catcgggcgt ttaccgtatc gcagaatggt ctgatatcac caacgcccac ggggttactg
     5521 gtgctggtat tgttgctggc ttgaaacaag gtgcgcaaga ggtcaccaaa gaaccaaggg
     5581 gattattgat gcttgctgaa ttgtcttcca agggttctct agcacacggt gaatatacta
     5641 agggtaccgt tgatattgca aagagtgata aagatttcgt tattgggttc attgctcaga
     5701 acgatatggg aggaagagaa gaagggtttg attggctaat catgacccca ggtgtaggtt
     5761 tagacgacaa aggcgatgca ttgggtcagc agtacagaac cgtcgacgaa gttgtaagtg
     5821 gtggatcaga tatcatcatt gttggcagag gacttttcgc caagggtaga gatcctaagg
     5881 ttgaaggtga aagatacaga aatgctggat gggaagcgta ccaaaagaga atcagcgctc
     5941 cccattaatt atacaggaaa cttaatagaa caaatcacat atttaatcta atagccacct
     6001 gcattggcac ggtgcaacac tacttcaact tcatcttaca aaaagatcac gtgatctgtt
     6061 gtattgggat ctctagacct aataacttcg tatagcatac att
//
