LOCUS       62056_14490        3418 bp DNA     circular SYN 27-AUG-2019
DEFINITION  Expression vector pLT_196, complete sequence.
ACCESSION   MK542538
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3418)
  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 3418)
  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 3418)
  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..3418
                     /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             1337..1354
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1388..1435
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1533..1562
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1659..2247
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2379..2406)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2433..3239)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(3240..3330)
                     /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 taaaaaatga aaatatttaa
      181 gttccatatg ccccccatta atttgatcgg ggtcggatgt cttaaagatg tgggaaggga
      241 aataaaaaaa ctgggtttca agaaaggtat tattgtaact gataaagttt tggtaagagc
      301 cgggcttgta aacaatgtta taagcgtgct ggaagaggaa ggaatcgaat atgttgtttt
      361 tgatgagaca aagccgaatc cgacaataaa gaatgtaacc aacggtttaa agctgttgat
      421 tgaaaataaa tgtgatttta ttatctcctg cggaggcgga agtgcccatg actgtgcaaa
      481 gggaatagga cttatcgcca aagagaaaaa ttttatagac gaagtggaaa ggcttgacaa
      541 ggtaaaatgc ggtggatgga attcggcttt gttactgcct ttggtggcaa ttaacacaac
      601 tgcaggaacc ggaagcgaag ttaccaaatt tgccataatt acagatgaag aaaagagaat
      661 taaaatgcct attgtggact ggagaataac tcctcttatt gccgtaaatg atccgctgct
      721 tatgataggt atgccgaaat ctttgacggc agcttccgga atggatgccc ttacccatgc
      781 cattgaagct tacataagta ttgacgcaaa tccttttacg gatgcacttg cattaaaagc
      841 aatagaaatc attttcaatt atttgaaaag agcggttgaa aacggaaatg acattgaggc
      901 aagagaaaaa atggcttatg ctgaattttt ggcaggtata gcttttaata atgcgggatt
      961 gggatatgtg catgcaatgg cacaccagct tggcggtttt tatgaccttc cccacggtgt
     1021 ttgcaatgct gttttacttc ctcatgttct tgaatacaac ttggaagcag tgcaaaacaa
     1081 gcttatatat attgcaaaag ctatgggaat tgatgttgac aaacttacaa caaaagaaat
     1141 tggcggcaaa attatagagt caataaacca gctgtcccag gaaataggca taccgtcacg
     1201 tctcaaagaa cttggagtaa aggaagaaga tattaaggag ctttcacaaa atgccctgaa
     1261 agacgtatgc ggctttacca atccaaaaaa ggcaactctt gaggatataa taaatatttt
     1321 taaaagcgcc atgggtcatc accaccacca ccattagggt ctcgcggccg ccaccgctga
     1381 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1441 aggaggaact atatccgggt aacgaattca agcttgatat cattcaggac gagcctcaga
     1501 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     1561 cggtagaagt cttctcatga ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac
     1621 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1681 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1741 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1801 actgttcttc tagtgtagcc gtagttagcc caccacttca agaactctgt agcaccgcct
     1861 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1921 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1981 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2041 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2101 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2161 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2221 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2281 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2341 aaccgtatta ccgcctttga gtgagctgat accgctcgcg cagaaaggcc cacccgaagg
     2401 tgagccaggt gattacattt gggccctcat tagaaaaact catcgagcat caaatgaaat
     2461 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     2521 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     2581 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     2641 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     2701 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     2761 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg
     2821 ttaaaaggac aattacaaac aggaatcgag tgcaaccggc gcaggaacac tgccagcgca
     2881 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaacgc tgtttttccg
     2941 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     3001 ggaagtggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     3061 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag
     3121 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     3181 tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata
     3241 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     3301 tgaaagttgg aacctcttac gtgccgatca agaagacggt caaaagcctc cggtcggagg
     3361 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagtatt gagcgata
//
