LOCUS       62056_14475        3028 bp DNA     circular SYN 27-AUG-2019
DEFINITION  Expression vector pLT_193, complete sequence.
ACCESSION   MK542535
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3028)
  AUTHORS   Tian L, Conway PM, Cervenka ND, Cui J, Maloney M, Olson DG, Lynd LR.
  TITLE     Metabolic engineering of Clostridium thermocellum for n-butanol 
            production from cellulose
  JOURNAL   Biotechnol Biofuels 12, 186 (2019)
  PUBMED    31367231
REFERENCE   2  (bases 1 to 3028)
  AUTHORS   Tian L.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-FEB-2019) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Dr, Hanover, New Hampshire 03755, United 
            States
REFERENCE   3  (bases 1 to 3028)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            Biofuels 12, 186 (2019)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-FEB-2019) Thayer School of Engineering, Dartmouth College, 14 
            Engineering Dr, Hanover, New Hampshire 03755, United States"
FEATURES             Location/Qualifiers
     source          1..3028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..134
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             947..964
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      998..1045
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1143..1172
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1269..1857
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1989..2016)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2043..2849)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(2850..2940)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tctagagaat tcgtccacca caattcagca aattgtgaac atcatcacgt tcatctttcc
       61 ctggttgcca atggcccatt ttcctgtcag taacgagaag gtcgcgaatt caggcgcttt
      121 ttagactggt cgtagagacc atgaaattct tttaaggagg taaaaaatgg attttaacaa
      181 tgtactttta aacaaggatg acggaattgc acttattata ataaatagac cgaaagcatt
      241 gaatgcttta aattatgaaa ctttgaagga acttgacagt gtgctggata ttgttgaaaa
      301 tgataaagaa ataaaagttt taattattac cggttccggt gaaaaagctt ttgttgccgg
      361 agccgacata gctgaaatga gcaatatgac acctttggaa gccaagaaat tttctcttta
      421 cggacaaaaa gtattcagaa aaatagaaat gctgtcaaag cctgtaattg cggcggtaaa
      481 cggttttgcc ttgggcggag gatgtgaatt gtcaatggca tgtgatataa gaatagctag
      541 caaaaatgca aaattcggac agccggaagt gggacttgga ataattccgg gattttccgg
      601 aacacaaaga cttcccaggc ttataggtac ttcaaaagcc aaagagctga tttttacagg
      661 agatatgata aacagtgatg aagcatataa aattggtttg ataagcaagg ttgtggagct
      721 ttccgacttg attgaagaag ctaaaaagct tgcaaaaaaa atgatgagta aaagccagat
      781 agcaatatcc ctggcaaagg aggcaattaa caaaggtatg gaaacggatt tggatacggg
      841 aaatacaatt gaggcggaaa aattttcgct ttgctttacc accgacgacc agaaagaagg
      901 aatgattgcc ttttcagaaa agagggctcc aaagtttgga aaaggtcatc accaccacca
      961 ccattagggt ctcgcggccg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1021 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccgggt aacgaattca
     1081 agcttgatat cattcaggac gagcctcaga ctccagcgta actggactgc aatcaactca
     1141 ctggctcacc ttcacgggtg ggcctttctt cggtagaagt cttctcatga ccaaaatccc
     1201 ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1261 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1321 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1381 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttagcc
     1441 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1501 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1561 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1621 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1681 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1741 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1801 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1861 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1921 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1981 accgctcgcg cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat
     2041 tagaaaaact catcgagcat caaatgaaat tgcaatttat tcatatcagg attatcaata
     2101 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     2161 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     2221 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     2281 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     2341 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     2401 gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgag
     2461 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     2521 tcttctaata cctggaacgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca
     2581 tcaggagtac ggataaaatg cttgatggtc ggaagtggca taaattccgt cagccagttt
     2641 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     2701 aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca
     2761 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     2821 ctcgacgttt cccgttgaat atggctcata gctcctgaaa atctcgataa ctcaaaaaat
     2881 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
     2941 agaagacggt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     3001 tgatttaaat ggtcagtatt gagcgata
//
