LOCUS       62056_14485        3649 bp DNA     circular SYN 27-AUG-2019
DEFINITION  Expression vector pLT_195, complete sequence.
ACCESSION   MK542537
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3649)
  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 3649)
  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 3649)
  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..3649
                     /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             1568..1585
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1619..1666
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1764..1793
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1890..2478
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2610..2637)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2664..3470)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(3471..3561)
                     /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 ttgacgaaaa
      181 cctagtagta accataacaa aaaaaatttt aaatgagata aacttaaaag aagccgagga
      241 aaaaaaggaa aaggataatc ccgaccttgg aatatttaat gatgtaaacg aagctgtcga
      301 atgtgccaaa gaggctcaaa agaagtttgc cttgatggat cttgaaaaaa gggaagaaat
      361 aatagcggct ataagggagg catgtgtaaa taatgctagg cttttggcgg aaatagcatg
      421 cagcgaaaca ggtcgtggaa gagttgagga taaagttgca aagaatatac tggccgcaaa
      481 aaaaactccg ggaactgaag atttgaagcc tacggcatgg acaggagaca ggggacttac
      541 tttggtggaa atggcaccag tcggagttat agcttcaatt acacctgtta ccaatcctac
      601 agcaactata attaacaaca ccatttcaat gcttgcggca ggcaatgccg ttgtgtttaa
      661 cccccatccg tcggctaaga aaacaagcaa caaagcggtg gagattataa atgaagcaat
      721 tttgaaagta ggcgccccca acggtcttgt ctgcagtatt aataatccaa caattcagac
      781 ggcccaaaaa ctgatggaac atcctgaagt taacatggtt gtagttacag gcggcaaagc
      841 cgtggttcag actgctttaa gatgcggaaa gaaagtgatt ggtgccggag caggaaatcc
      901 gccggtggta gtggatgaaa ccgccgatat tgtaaaggct gcacatgaca tagcctgcgg
      961 tgcgagcttt gacaacaatc ttccgtgtat tgcagagaaa gaaattattg cggttgaaag
     1021 aattgccgac acattgctgg aaaggatgaa aagagaaggt gcatatgttc tgcacggtaa
     1081 ggacatagac agaatgacgg aacttatttt tcagggagga gccataaata aagacttaat
     1141 tggaagggat gcccatttta ttctgtccca gataggtatt gaaacgggaa aagatataag
     1201 acttgttgtt atgcctgtag atgtttccca tccccttgta tatcatgaac agcttatgcc
     1261 ggtaatacct tttgtaactg ttccgacagt tgaagaggcc atcaatttgg ctgttaaagc
     1321 agaaggcgga aacaggcaca ctgcaatgat gcattcaaaa aacgtggaaa atatgaccgc
     1381 ttttgcaagg gcaatacaga caacaatatt cgtaaaaaat gcaccttctt atgcaggtat
     1441 aggatttgga ggtgaaggat acaccacttt taccatcgcg ggaccgaccg gagaagggct
     1501 tacctctgca agaaccttta caagacaaag aagatgtgtt ttggttgatg ctttcagaat
     1561 agtaggtcat caccaccacc accattaggg tctcgcggcc gccaccgctg agcaataact
     1621 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1681 tatatccggg taacgaattc aagcttgata tcattcagga cgagcctcag actccagcgt
     1741 aactggactg caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaag
     1801 tcttctcatg accaaaatcc cttaacgtga gttacgcgcg cgtcgttcca ctgagcgtca
     1861 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1921 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1981 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2041 ctagtgtagc cgtagttagc ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2101 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2161 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2221 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2281 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2341 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2401 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2461 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2521 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2581 accgcctttg agtgagctga taccgctcgc gcagaaaggc ccacccgaag gtgagccagg
     2641 tgattacatt tgggccctca ttagaaaaac tcatcgagca tcaaatgaaa ttgcaattta
     2701 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     2761 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     2821 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     2881 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     2941 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     3001 ccgttattca ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga
     3061 caattacaaa caggaatcga gtgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     3121 ttttcacctg aatcaggata ttcttctaat acctggaacg ctgtttttcc ggggatcgca
     3181 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagtggc
     3241 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     3301 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt
     3361 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     3421 atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat agctcctgaa
     3481 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     3541 gaacctctta cgtgccgatc aagaagacgg tcaaaagcct ccggtcggag gcttttgact
     3601 ttctgctatg gaggtcaggt atgatttaaa tggtcagtat tgagcgata
//
