LOCUS       .                       6305 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        97..131
                     /label="J23119(SpeI) promoter"
                     /note="bacterial promoter (Registry of Standard Biological
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        149..224
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     primer_bind     complement(257..273)
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(332..348)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(356..372)
                     /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)."
     promoter        complement(380..410)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(425..446)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(734..1322)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(1496..2308)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(2309..2413)
                     /label="AmpR promoter"
     primer_bind     2887..2903
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(2919..3941)
                     /label="HygR"
                     /note="hygromycin B phosphotransferase"
     promoter        complement(4397..4499)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     primer_bind     4517..4533
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(4598..6016)
                     /label="SacB"
                     /note="secreted levansucrase that renders bacterial growth
                     sensitive to sucrose"
ORIGIN
        1 cgggcctaaa gaactaaaaa atctatctgt ttcttttcat tctctgtatt ttttatagtt
       61 tctgttgcat gggcataaag ttgcaagctt gatatcttga cagctagctc agtcctaggt
      121 ataatactag tcgagaccat tggtctcagt tttagagcta gaaatagcaa gttaaaataa
      181 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgatatcgaa
      241 ttcctgcagc ccgggggatc cactagttct agagcggccg ccaccgcggt ggagctcatc
      301 ccaatggcgc gccgagcttg gctcgagcat ggtcatagct gtttcctgtg tgaaattgtt
      361 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      421 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      481 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
      541 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      901 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1141 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1441 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttagaaaa
     1501 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1561 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1621 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     1681 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     1741 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     1801 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     1861 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     1921 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     1981 atacctggaa tgctgttttc ccagggatcg cagtggtgag taaccatgca tcatcaggag
     2041 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     2101 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     2161 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     2221 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     2281 aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt
     2341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     2461 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     2521 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2581 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2641 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     2701 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     2761 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     2821 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     2881 gacgttgtaa aacgacggcc agtgaattga cgcgtctatt cctttgccct cggacgagtg
     2941 ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc
     3001 gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg
     3061 tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag
     3121 agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc
     3181 ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag
     3241 aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg
     3301 accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg
     3361 aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg
     3421 acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca
     3481 atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg
     3541 ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc
     3601 ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca
     3661 cggcgggaga tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga
     3721 atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg
     3781 gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga
     3841 gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga
     3901 aacttctcga cagacgtcgc ggtgagttca ggctttttca tcagtcgatc atagcacgat
     3961 caacggcact gttgcaaata gtcggtggtg ataaacttat catccccttt tgctgatgga
     4021 gctgcacatg aacccattca aaggccggca ttttcagcgt gacatcattc tgtgggccgt
     4081 acgctggtac tgcaaatacg gcatcagtta ccgtgagccg gatcagtgag ggtttgcaac
     4141 tgcgggtcaa ggatctggat ttcgatcacg gcacgatcat cgtgcgggag ggcaagggct
     4201 ccaaggatcg ggccttgatg ttacccgaga gcttggcacc cagcctgcgc gagcagggga
     4261 attgatccgg tggatgaccc agctgaacgg tctggttata ggtacattga gcaactgact
     4321 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     4381 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     4441 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcag
     4501 gtaccgggcc ccccctcgag gtcgacggta tcgatacggc atcagagcag attgtactga
     4561 gagtgcacca taatcggcat tttcttttgc gtttttattt gttaactgtt aattgtcctt
     4621 gttcaaggat gctgtctttg acaacagatg ttttcttgcc tttgatgttc agcaggaagc
     4681 taggcgcaaa cgttgattgt ttgtctgcgt agaatcctct gtttgtcata tagcttgtaa
     4741 tcacgacatt gtttcctttc gcttgaggta cagcgaagtg tgagtaagta aaggttacat
     4801 cgttaggatc aagatccatt tttaacacaa ggccagtttt gttcagcggc ttgtatgggc
     4861 cagttaaaga attagaaaca taaccaagca tgtaaatatc gttagacgta atgccgtcaa
     4921 tcgtcatttt tgatccgcgg gagtcagtga acagatacca tttgccgttc attttaaaga
     4981 cgttcgcgcg ttcaatttca tctgttactg tgttagatgc aatcagcggt ttcatcactt
     5041 ttttcagtgt gtaatcatcg tttagctcaa tcataccgag agcgccgttt gctaactcag
     5101 ccgtgcgttt tttatcgctt tgcagaagtt tttgactttc ttgacggaag aatgatgtgc
     5161 ttttgccata gtatgctttg ttaaataaag attcttcgcc ttggtagcca tcttcagttc
     5221 cagtgtttgc ttcaaatact aagtatttgt ggcctttatc ttctacgtag tgaggatctc
     5281 tcagcgtatg gttgtcgcct gagctgtagt tgccttcatc gatgaactgc tgtacatttt
     5341 gatacgtttt tccgtcaccg tcaaagattg atttataatc ctctacaccg ttgatgttca
     5401 aagagctgtc tgatgctgat acgttaactt gtgcagttgt cagtgtttgt ttgccgtaat
     5461 gtttaccgga gaaatcagtg tagaataaac ggatttttcc gtcagatgta aatgtggctg
     5521 aacctgacca ttcttgtgtt tggtctttta ggatagaatc atttgcatcg aatttgtcgc
     5581 tgtctttaaa gacgcggcca gcgtttttcc agctgtcaat agaagtttcg ccgacttttt
     5641 gatagaacat gtaaatcgat gtgtcatccg catttttagg atctccggct aatgcaaaga
     5701 cgatgtggta gccgtgatag tttgcgacag tgccgtcagc gttttgtaat ggccagctgt
     5761 cccaaacgtc caggcctttt gcagaagaga tatttttaat tgtggacgaa tcgaactcag
     5821 gaacttgata tttttcattt ttttgctgtt cagggatttg cagcatatca tggcgtgtaa
     5881 tatgggaaat gccgtatgtt tccttatatg gcttttggtt cgtttctttc gcaaacgctt
     5941 gagttgcgcc tcctgccagc agtgcggtag taaaggttaa tactgttgct tgttttgcaa
     6001 actttttgat gttcatcgtt catgtctcct tttttatgta ctgtgttagc ggtctgcttc
     6061 ttccagccct cctgtttgaa gatggcaagt tagttacgca caataaaaaa agacctaaaa
     6121 tatgtaaggg gtgacgccaa agtatacact ttgcccttta cacattttag gtcttgcctg
     6181 ctttatcagt aacaaacccg cgcgatttac ttttcgacct cattctatta gactctcgtt
     6241 tggattgcaa ctggtctatt ttcctctttt gtttgataga aaatcataaa aggatttgca
     6301 gacta
//
