LOCUS       40924_26476        3038 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pK18sB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3038)
  AUTHORS   Jayakody LN, Johnson CW, Whitham JM, Giannone RJ, Black BA, 
            Cleveland NS, Klingeman DM, Michener WE, Olstad JL, Vardon DR, Brown
            RC, Brown SD, Hettich RL, Guss AM, Beckham GT.
  TITLE     Thermochemical wastewater valorization via enhanced microbial 
            toxicity tolerance
  JOURNAL   Energy Environ Sci (2018) In press
REFERENCE   2  (bases 1 to 3038)
  AUTHORS   Johnson CW.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-APR-2018) National Bioenergy Center, National 
            Renewable Energy Laboratory, 15301 Denver West Parkway, Golden, CO 
            80401, USA
REFERENCE   3  (bases 1 to 3038)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3038)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Energy 
            Environ Sci (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-APR-2018) National Bioenergy Center, National Renewable Energy 
            Laboratory, 15301 Denver West Parkway, Golden, CO 80401, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     601..617
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    626..682
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(686..702)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      713..762
                     /label=NEOKAN promoter
                     /note="NEOKAN promoter"
                     /regulatory_class="promoter"
     CDS             784..1575
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             1585..3003
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgcctcaca
      601 caggaaacag ctatgacatg attacgaatt cgagctcggt acccggggat cctctagagt
      661 cgacctgcag gcatgcaagc ttggcactgg ccgtcgtttt acaacgtcgt gaccggaatt
      721 gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaacaggatg aggatcgttt
      781 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
      841 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
      901 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
      961 ctccaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     1021 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     1081 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     1141 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     1201 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     1261 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcggatgccc
     1321 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     1381 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     1441 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     1501 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     1561 cttgacgagt tcttctgagc gacgatgaac atcaaaaagt ttgcaaaaca agcaacagta
     1621 ttaaccttta ctaccgcact gctggcagga ggcgcaactc aagcgtttgc gaaagaaacg
     1681 aaccaaaagc catataagga aacatacggc atttcccata ttacacgcca tgatatgctg
     1741 caaatccctg aacagcaaaa aaatgaaaaa tatcaagttt ctgaatttga ttcgtccaca
     1801 attaaaaata tctcttctgc aaaaggcctg gacgtttggg acagctggcc attacaaaac
     1861 gctgacggca ctgtcgcaaa ctatcacggc taccacatcg tctttgcatt agccggagat
     1921 cctaaaaatg cggatgacac atcgatttac atgttctatc aaaaagtcgg cgaaacttct
     1981 attgacagct ggaaaaacgc tggccgcgtc tttaaagaca gcgacaaatt cgatgcaaat
     2041 gattctatcc taaaagacca aacacaagaa tggtcaggtt cagccacatt tacatctgac
     2101 ggaaaaatcc gtttattcta cactgatttc tccggtaaac attacggcaa acaaacactg
     2161 acaactgcac aagttaacgt atcagcatca gacagctctt tgaacatcaa cggtgtagag
     2221 gattataaat caatctttga cggtgacgga aaaacgtatc aaaatgtaca gcagttcatc
     2281 gatgaaggca actacagctc aggcgacaac catacgctga gagatcctca ctacgtagaa
     2341 gataaaggcc acaaatactt agtatttgaa gcaaacactg gaactgaaga tggctaccaa
     2401 ggcgaagaat ctttatttaa caaagcatac tatggcaaaa gcacatcatt cttccgtcaa
     2461 gaaagtcaaa aacttctgca aagcgataaa aaacgcacgg ctgagttagc aaacggcgct
     2521 ctcggtatga ttgagctaaa cgatgattac acactgaaaa aagtgatgaa accgctgatt
     2581 gcatctaaca cagtaacaga tgaaattgaa cgcgcgaacg tctttaaaat gaacggcaaa
     2641 tggtacctgt tcactgactc ccgcggatca aaaatgacga ttgacggcat tacgtctaac
     2701 gatatttaca tgcttggtta tgtttctaat tctttaactg gcccatacaa gccgctgaac
     2761 aaaactggcc ttgtgttaaa aatggatctt gatcctaacg atgtaacctt tacttactca
     2821 cacttcgctg tacctcaagc gaaaggaaac aatgtcgtga ttacaagcta tatgacaaac
     2881 agaggattct acgcagacaa acaatcaacg tttgcgccga gcttcctgct gaacatcaaa
     2941 ggcaagaaaa catctgttgt caaagacagc atccttgaac aaggacaatt aacagttaac
     3001 aaataatcag accccgtaga aaagatcaaa ggatcttc
//
