LOCUS       62056_19675        4704 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4704)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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-beta-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)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        complement(2297..2388)
                     /label=AmpR promoter
     primer_bind     2862..2878
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2916..2932
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2997..4415)
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
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 agtcaacccc
     1501 aagtcagagg gtccaatcgc agatagaagg caaggcgttc gcggtcgggg gcttcgatcc
     1561 ccaatacatt gaataggaca gcgaaggcgc gctctgcttc atctggcgct gcccagttct
     1621 cttcggcgtt agcaatcatg agtgccaaat cggcatagcg atctgctgtt ccgagccgcc
     1681 caaggtcgat cagacccgtg cattgaagag ttttagggtc caccatgaag ttcggcatgc
     1741 agggatcacc atggcaaaca accatatcgg tgcgctcttg gtcgagccgc accggtagct
     1801 ctcgttcgac acgagccaaa agatcgagct gcggcgtact cttgtcctcg tccggtaaga
     1861 agtcgggatt gacggcattg cgggacacca catcaacggc gcgtccgaac attcgcgaca
     1921 gcctgcgctc aaacggacat tgatcaacca ataggctgtg aacagcgcca agttgctgcc
     1981 ccattgacgg ccacgctttg agcaaatccg ctccagacag atcagccgcc ggtactcccg
     2041 gaattgccgt tatcaccaag catgcaccct cctgttcctc ctgccagttg atcacctcgg
     2101 ggcaagccac acctcgacct ttgagccaaa tgaggcggtc acgctctcca gcgagctcac
     2161 cgcggcggga agcaggtgcg attttcgcga aggcatgccc gtcaccacgt cgaaaaacaa
     2221 aatcaccaga ttctccgcct ctgacaggca accagtcaga atgcgattca ccaaaaaaaa
     2281 tattagttcg attcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaaca catttccccg
     2401 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     2461 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2521 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2581 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2641 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2701 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     2761 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     2821 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     2881 attgacgcgt attgggatgg taccgggccc cccctcgagg tcgacggtat cgatacggca
     2941 tcagagcaga ttgtactgag agtgcaccat aatcggcatt ttcttttgcg tttttatttg
     3001 ttaactgtta attgtccttg ttcaaggatg ctgtctttga caacagatgt tttcttgcct
     3061 ttgatgttca gcaggaagct aggcgcaaac gttgattgtt tgtctgcgta gaatcctctg
     3121 tttgtcatat agcttgtaat cacgacattg tttcctttcg cttgaggtac agcgaagtgt
     3181 gagtaagtaa aggttacatc gttaggatca agatccattt ttaacacaag gccagttttg
     3241 ttcagcggct tgtatgggcc agttaaagaa ttagaaacat aaccaagcat gtaaatatcg
     3301 ttagacgtaa tgccgtcaat cgtcattttt gatccgcggg agtcagtgaa cagataccat
     3361 ttgccgttca ttttaaagac gttcgcgcgt tcaatttcat ctgttactgt gttagatgca
     3421 atcagcggtt tcatcacttt tttcagtgtg taatcatcgt ttagctcaat cataccgaga
     3481 gcgccgtttg ctaactcagc cgtgcgtttt ttatcgcttt gcagaagttt ttgactttct
     3541 tgacggaaga atgatgtgct tttgccatag tatgctttgt taaataaaga ttcttcgcct
     3601 tggtagccat cttcagttcc agtgtttgct tcaaatacta agtatttgtg gcctttatct
     3661 tctacgtagt gaggatctct cagcgtatgg ttgtcgcctg agctgtagtt gccttcatcg
     3721 atgaactgct gtacattttg atacgttttt ccgtcaccgt caaagattga tttataatcc
     3781 tctacaccgt tgatgttcaa agagctgtct gatgctgata cgttaacttg tgcagttgtc
     3841 agtgtttgtt tgccgtaatg tttaccggag aaatcagtgt agaataaacg gatttttccg
     3901 tcagatgtaa atgtggctga acctgaccat tcttgtgttt ggtcttttag gatagaatca
     3961 tttgcatcga atttgtcgct gtctttaaag acgcggccag cgtttttcca gctgtcaata
     4021 gaagtttcgc cgactttttg atagaacatg taaatcgatg tgtcatccgc atttttagga
     4081 tctccggcta atgcaaagac gatgtggtag ccgtgatagt ttgcgacagt gccgtcagcg
     4141 ttttgtaatg gccagctgtc ccaaacgtcc aggccttttg cagaagagat atttttaatt
     4201 gtggacgaat cgaactcagg aacttgatat ttttcatttt tttgctgttc agggatttgc
     4261 agcatatcat ggcgtgtaat atgggaaatg ccgtatgttt ccttatatgg cttttggttc
     4321 gtttctttcg caaacgcttg agttgcgcct cctgccagca gtgcggtagt aaaggttaat
     4381 actgttgctt gttttgcaaa ctttttgatg ttcatcgttc atgtctcctt ttttatgtac
     4441 tgtgttagcg gtctgcttct tccagccctc ctgtttgaag atggcaagtt agttacgcac
     4501 aataaaaaaa gacctaaaat atgtaagggg tgacgccaaa gtatacactt tgccctttac
     4561 acattttagg tcttgcctgc tttatcagta acaaacccgc gcgatttact tttcgacctc
     4621 attctattag actctcgttt ggattgcaac tggtctattt tcctcttttg tttgatagaa
     4681 aatcataaaa ggatttgcag acta
//
