LOCUS       V015087                 7032 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V015087
VERSION     V015087
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7032)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     475..491
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        498..516
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(607..2025)
                     /label="SacB"
                     /note="secreted levansucrase that renders bacterial growth
                     sensitive to sucrose"
     promoter        2487..2516
                     /label="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_RNA        2539..2614
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        complement(2694..2712)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2730..2746)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(2754..2770)
                     /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(2778..2808)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2823..2844)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3326..4060
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     rep_origin      complement(4401..4752)
                     /direction=LEFT
                     /label="pRO1600 oriV"
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             4766..5587
                     /label="pRO1600 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      6056..6644
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     rep_origin      complement(join(6911..7032,1..334))
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
       61 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
      121 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
      181 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
      241 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
      301 cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact
      361 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      421 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      481 cgacggccag tgaattgtaa tacgactcac tatagggcga attcgagctc ggtacccggg
      541 gatcccggca tcagagcaga ttgtactgag agtgcaccat aatcggcatt ttcttttgcg
      601 tttttatttg ttaactgtta attgtccttg ttcaaggatg ctgtctttga caacagatgt
      661 tttcttgcct ttgatgttca gcaggaagct aggcgcaaac gttgattgtt tgtctgcgta
      721 gaatcctctg tttgtcatat agcttgtaat cacgacattg tttcctttcg cttgaggtac
      781 agcgaagtgt gagtaagtaa aggttacatc gttaggatca agatccattt ttaacacaag
      841 gccagttttg ttcagcggct tgtatgggcc agttaaagaa ttagaaacat aaccaagcat
      901 gtaaatatcg ttagacgtaa tgccgtcaat cgtcattttt gatccgcggg agtcagtgaa
      961 cagataccat ttgccgttca ttttaaagac gttcgcgcgt tcaatttcat ctgttactgt
     1021 gttagatgca atcagcggtt tcatcacttt tttcagtgtg taatcatcgt ttagctcaat
     1081 cataccgaga gcgccgtttg ctaactcagc cgtgcgtttt ttatcgcttt gcagaagttt
     1141 ttgactttct tgacggaaga atgatgtgct tttgccatag tatgctttgt taaataaaga
     1201 ttcttcgcct tggtagccat cttcagttcc agtgtttgct tcaaatacta agtatttgtg
     1261 gcctttatct tctacgtagt gaggatctct cagcgtatgg ttgtcgcctg agctgtagtt
     1321 gccttcatcg atgaactgct gtacattttg atacgttttt ccgtcaccgt caaagattga
     1381 tttataatcc tctacaccgt tgatgttcaa agagctgtct gatgctgata cgttaacttg
     1441 tgcagttgtc agtgtttgtt tgccgtaatg tttaccggag aaatcagtgt agaataaacg
     1501 gatttttccg tcagatgtaa atgtggctga acctgaccat tcttgtgttt ggtcttttag
     1561 gatagaatca tttgcatcga atttgtcgct gtctttaaag acgcggccag cgtttttcca
     1621 gctgtcaata gaagtttcgc cgactttttg atagaacatg taaatcgatg tgtcatccgc
     1681 atttttagga tctccggcta atgcaaagac gatgtggtag ccgtgatagt ttgcgacagt
     1741 gccgtcagcg ttttgtaatg gccagctgtc ccaaacgtcc aggccttttg cagaagagat
     1801 atttttaatt gtggacgaat cgaactcagg aacttgatat ttttcatttt tttgctgttc
     1861 agggatttgc agcatatcat ggcgtgtaat atgggaaatg ccgtatgttt ccttatatgg
     1921 cttttggttc gtttctttcg caaacgcttg agttgcgcct cctgccagca gtgcggtagt
     1981 aaaggttaat actgttgctt gttttgcaaa ctttttgatg ttcatcgttc atgtctcctt
     2041 ttttatgtac tgtgttagcg gtctgcttct tccagccctc ctgtttgaag atggcaagtt
     2101 agttacgcac aataaaaaaa gacctaaaat atgtaagggg tgacgccaaa gtatacactt
     2161 tgccctttac acattttagg tcttgcctgc tttatcagta acaaacccgc gcgatttact
     2221 tttcgacctc attctattag actctcgttt ggattgcaac tggtctattt tcctcttttg
     2281 tttgatagaa aatcataaaa ggatttgcag actacgggcc taaagaacta aaaaatctat
     2341 ctgtttcttt tcattctctg tattttttat agtttctgtt gcatgggcat aaagttgcgt
     2401 cgacgccttg ctcaaattgg aatcaggttt gtgccaatac cagtagaaac agacgaagaa
     2461 tccatggcgg ccgctgtcat gggcccttga caattaatca tccggctcgt ataatgtgtg
     2521 gcgagaccat tggtctcagt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
     2581 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgagatctgt ccatacccat
     2641 ggtctagaat gctcgagact tttcatactc ccgccattca gaagcttgag tattctatag
     2701 tgtcacctaa atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2761 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2821 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2881 aacctgtcgt gccagaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     2941 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     3001 ccttagaagc aaacttagag tgtgttgata gtgcattatc ttaaaatttt gtataatagg
     3061 aattgaagtt aaattagatg ctaaaaatag gaattgaagt taaattagat gctaaaaatt
     3121 tgtaattaag aaggagggat tcgtcatgtt ggtattccaa atgcgtaatg tagataaaac
     3181 atctactgtt ttgaaacaga ctaaaaacag tgattacgca gataaataaa tacgttagat
     3241 taattcctac cagtgactaa tcttatgact ttttaaacag ataactaaaa ttacaaacaa
     3301 atcgtttaac ttcaggagag attacatgaa caaaaatata aaatattctc aaaacttttt
     3361 aacgagtgaa aaagtactca accaaataat aaaacaattg aatttaaaag aaaccgatac
     3421 cgtttacgaa attggaacag gtaaagggca tttaacgacg aaactggcta aaataagtaa
     3481 acaggtaacg tctattgaat tagacagtca tctattcaac ttatcgtcag aaaaattaaa
     3541 actgaatact cgtgtcactt taattcacca agatattcta cagtttcaat tccctaacaa
     3601 acagaggtat aaaattgttg ggaatattcc ttacaattta agcacacaaa ttattaaaaa
     3661 agtggttttt gaaagccgtg cgtctgacat ctatctgact gttgaagaag gattctacaa
     3721 gcgtaccttg gatattcacc gaacactagg gttgctcttg cacactcaag tctcgattca
     3781 gcaattgctt aagctgccag cggaatgctt tcatcctaaa ccaaaagtaa acagtgtctt
     3841 aataaaactt acccgccata ccacagatgt tccagataaa tattggaagc tatataagta
     3901 ctttgtttca aaatgggtca atcgagaata tcgtcaactg tttactaaaa atcagtttcg
     3961 tcaagcaatg aaacacgcca aagtaaacaa tttaagtacc attacttatg agcaagtatt
     4021 gtctattttt aatagttatc tattatttaa cgggaggaaa taactgtcag accaagttta
     4081 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4141 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4201 gtcagacccc aattacacgc cactggctgt gcttgctggg gtgacggtgg caacggtggc
     4261 ggccttgctg ggctatcgcg ttggaaagaa acgagggaaa ggggactgat aaaccggtct
     4321 tagcccctcc ccttggtgtc caaccgctct gtaggcctct caggcgccgc tggtgccgct
     4381 ggttggacgc caagggtgaa tccgcctcga taccctgatt actcgcttcc tgcgccctct
     4441 caggcggcga taggggactg gtaaaacggg gattgcccag acgcctcccc cgccccttca
     4501 ggggcacaaa tgcggcccca acggggccac gtagtggtgc gttttttgcg tttccaccct
     4561 tttcttcctt ttccctttta aaccttttag gacgtctaca ggccacgtaa tccgtggcct
     4621 gtagagttta aaaagggacg gatttgttgc cattaaggga cggatttgtt gttaagaagg
     4681 gacggatttg ttgttgtaaa gggacggatt tgttgtattg tgggacgcag atacagtgtc
     4741 cccttataca caaggaatgt cgaacgtggc ctcaccccca atggtttaca aaagcaatgc
     4801 cctggtcgag gccgcgtatc gcctcagtgt tcaggaacag cggatcgttc tggcctgtat
     4861 tagccaggtg aagaggagcg agcctgtcac cgatgaagtg atgtattcag tgacggcgga
     4921 ggacatagcg acgatggcgg gtgtccctat cgaatcttcc tacaaccagc tcaaagaagc
     4981 ggccctgcgc ctgaaacggc gggaagtccg gttaacccaa gagcccaatg gcaaggggaa
     5041 aagaccgagt gtgatgatta ccggctgggt gcaaacaatc atctaccggg agggtgaggg
     5101 ccgtgtagaa ctcaggttca ccaaagacat gctgccgtac ctgacggaac tcaccaaaca
     5161 gttcaccaaa tacgccttgg ctgacgtggc caagatggac agcacccacg cgatcaggct
     5221 ttacgagctg ctcatgcaat gggacagcat cggccagcgc gaaatagaaa ttgaccagct
     5281 gcgaaagtgg tttcaactgg aaggccggta tccctcgatc aaggacttca agttgcgagt
     5341 gcttgatcca gccgtgacgc agatcaacga gcacagcccg ctacaggtgg agtgggcgca
     5401 gcgaaagacc gggcgcaagg tcacacatct gttgttcagt tttggaccga agaagcccgc
     5461 caaggcggtg ggtaaggccc cagcgaagcg caaggccggg aagatttcag atgctgagat
     5521 cgcgaaacag gctcgccctg gtgagacatg ggaagcggcc cgcgctcgac taacccagat
     5581 gccgcttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5641 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5701 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5761 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5821 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5881 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5941 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6001 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6061 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6181 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6721 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6781 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     6841 attttctcct tacgcatctg tgcggtattt cacaccgcac agatgcgtaa ggagaaaata
     6901 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6961 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     7021 tagaccgaga ta
//
