LOCUS       62056_14480        3427 bp DNA     circular SYN 27-AUG-2019
DEFINITION  Expression vector pLT_194, complete sequence.
ACCESSION   MK542536
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3427)
  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 3427)
  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 3427)
  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..3427
                     /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             1346..1363
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1397..1444
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1542..1571
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1668..2256
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2388..2415)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2442..3248)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(3249..3339)
                     /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 taataaaacc
      181 catgataaga aacaatatat gtatgaatgc ccatcccgaa ggctgcagac tgtttgtaga
      241 caggcagatt gattatgtaa gacaaagagg tccgttatca tccggaccca aaaatgtttt
      301 ggttgtggga agcagtggcg gatacggttt ggccacaaga attgcagccg gattcggagc
      361 aggtgcagca actatcggag tatattttga gaaggagcct agtgaaaaaa gaacaggaac
      421 accgggatgg tatgcggcaa ggcatttttt gaagagagcc gaggctgcgg ggctgaaggt
      481 ccatgacata aacggagatg ctttcagctt tgaagttaaa gaacaggtgg ttgatttaat
      541 caaaaaagaa tatggtacca tagatttgtt tgtttattct ttggcttcgg gagttcgcac
      601 cgaccctgta agcggcgtga catacagaag tgcactgaag cctattggaa gaacctatac
      661 ttcaaaggcg cttgacccgt taaagaggga agtaattcag gcaaccattg aaccggcatc
      721 tgaggaagaa attgaggcaa cagtgaaagt tatgggagga gaagactggc agctttgggt
      781 tgaagcatta aaagaaggag gtgttcttgc agagggtgcc gttactgttg cttattcata
      841 tataggcccg gaacttacca gaccgatata tcgtgagggc acaataggaa aggctaaaga
      901 gcatcttgaa gcgacggcaa aaacacttga tgaagctttg gcatccgtgg gtggaaaagc
      961 atatgtttcg gtaaataaag ctgtggttac aagggcttca gctgttattc ctgcagtacc
     1021 tttgtatctt tccattcttt ttaccgtaat gaaagccaaa ggaattcatg aaggatgtat
     1081 agaacaaatg tatagaatgc ttgccgagag gctttatgcc ggagggcctg tgcctgttga
     1141 cgatgaggga aggataagga tggatgattg ggaaatgaga gaagatgttc aaagggaggt
     1201 tgcagaagta tgggaaagaa taactacgga aacactggac agagaagccg atgtggacgg
     1261 atatttgacg gattttttaa accttcacgg tttcggtttt aaagagattg actatgaaaa
     1321 ggaagtggat ccacgggaag gtggtcatca ccaccaccac cattagggtc tcgcggccgc
     1381 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1441 tttgctgaaa ggaggaacta tatccgggta acgaattcaa gcttgatatc attcaggacg
     1501 agcctcagac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg
     1561 gcctttcttc ggtagaagtc ttctcatgac caaaatccct taacgtgagt tacgcgcgcg
     1621 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1681 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1741 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1801 ataccaaata ctgttcttct agtgtagccg tagttagccc accacttcaa gaactctgta
     1861 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1921 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1981 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2041 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2101 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2161 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2221 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2281 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2341 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgcgc agaaaggccc
     2401 acccgaaggt gagccaggtg attacatttg ggccctcatt agaaaaactc atcgagcatc
     2461 aaatgaaatt gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     2521 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     2581 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     2641 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     2701 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     2761 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag gcgaaatacg
     2821 cgatcgctgt taaaaggaca attacaaaca ggaatcgagt gcaaccggcg caggaacact
     2881 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaacgct
     2941 gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     3001 ttgatggtcg gaagtggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     3061 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     3121 ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     3181 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc ccgttgaata
     3241 tggctcatag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     3301 tcattatggt gaaagttgga acctcttacg tgccgatcaa gaagacggtc aaaagcctcc
     3361 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagtattg
     3421 agcgata
//
