LOCUS       62056_14470        3094 bp DNA     circular SYN 27-AUG-2019
DEFINITION  Expression vector pLT_190, complete sequence.
ACCESSION   MK542534
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3094)
  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 3094)
  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 3094)
  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..3094
                     /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             1013..1030
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1064..1111
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1209..1238
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1335..1923
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2055..2082)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2109..2915)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(2916..3006)
                     /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 taaaaaatgc aaaaaatatg
      181 tgtaataggg gcgggaacaa tgggaagcgg tattgcccag gtttttgcac aaaacggctt
      241 cgaagtaatt ttaagggata ttgacatgaa gtttgtagag aaaggctttg gaacgattga
      301 aaaaaatctt caaagaaatg ttgacaaagg aaagattacc gccgacgaaa agaatgaaat
      361 cctgtccagg ataagaggta caacaaacct tgaagatgca aaagaagcgg attttgttgt
      421 ggaagctgcg atagaaaaca tggatttgaa aaaacagata tttaaagagt tggatgaaat
      481 ttgcaaaatg gaaaccattc ttgcttccaa tacttcgagt ctctcaataa cggaaatagc
      541 ttctgccaca aaaagaccgg aaaaagttat tggaatgcat tttttcaatc ctgttccggt
      601 aatgaaactg gttgaagtta taaaagggct gaaaacaagt gaacagactt ttaatgtggt
      661 gagggaactt gcattaaaag tggacaagac accgattgag gtaaaggaag ccccgggatt
      721 tgtggtaaac agaattttga ttcccatgat aaacgaagca ataggaatac tggcagacgg
      781 agttgcaacc gcggaagaca ttgatgaggc aatgaaactt ggagcaaatc atccgatagg
      841 tcctttggct ttaagcgatt taatcggaaa tgatgttgtt cttgccataa tgaatgtact
      901 ttatgaagag tatggagatt caaaatacag acctcaccct ctgcttaaaa agatggtaag
      961 aggaggactt ttgggaagaa aaaccggaaa gggtttcttt gaatataaag gtcatcacca
     1021 ccaccaccat tagggtctcg cggccgccac cgctgagcaa taactagcat aaccccttgg
     1081 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccgggtaacg
     1141 aattcaagct tgatatcatt caggacgagc ctcagactcc agcgtaactg gactgcaatc
     1201 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaagtcttc tcatgaccaa
     1261 aatcccttaa cgtgagttac gcgcgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     1321 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1381 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1441 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1501 ttagcccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1561 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1621 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1681 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1741 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1801 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1861 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1921 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1981 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2041 gctgataccg ctcgcgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc
     2101 cctcattaga aaaactcatc gagcatcaaa tgaaattgca atttattcat atcaggatta
     2161 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     2221 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     2281 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     2341 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     2401 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     2461 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     2521 atcgagtgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     2581 ggatattctt ctaatacctg gaacgctgtt tttccgggga tcgcagtggt gagtaaccat
     2641 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gtggcataaa ttccgtcagc
     2701 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2761 agaaacaact ctggcgcatc gggcttccca tacaagcgat agattgtcgc acctgattgc
     2821 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2881 cgcggcctcg acgtttcccg ttgaatatgg ctcatagctc ctgaaaatct cgataactca
     2941 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     3001 cgatcaagaa gacggtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     3061 caggtatgat ttaaatggtc agtattgagc gata
//
