LOCUS       62056_14455        3427 bp DNA     circular SYN 27-AUG-2019
DEFINITION  Expression vector pLT_181, complete sequence.
ACCESSION   MK542531
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"
     rep_origin      39..627
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(759..786)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(813..1619)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(1620..1710)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     promoter        1814..1932
                     /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             3144..3161
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3195..3242
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3340..3369
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
       61 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      181 actgttcttc tagtgtagcc gtagttagcc caccacttca agaactctgt agcaccgcct
      241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
      301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
      361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
      421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      721 aaccgtatta ccgcctttga gtgagctgat accgctcgcg cagaaaggcc cacccgaagg
      781 tgagccaggt gattacattt gggccctcat tagaaaaact catcgagcat caaatgaaat
      841 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
      901 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
      961 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1021 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     1081 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1141 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg
     1201 ttaaaaggac aattacaaac aggaatcgag tgcaaccggc gcaggaacac tgccagcgca
     1261 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaacgc tgtttttccg
     1321 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1381 ggaagtggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1441 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag
     1501 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1561 tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata
     1621 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     1681 tgaaagttgg aacctcttac gtgccgatca agaagacggt caaaagcctc cggtcggagg
     1741 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagtatt gagcgatatc
     1801 tagagaattc gtccaccaca attcagcaaa ttgtgaacat catcacgttc atctttccct
     1861 ggttgccaat ggcccatttt cctgtcagta acgagaaggt cgcgaattca ggcgcttttt
     1921 agactggtcg tagagaccat gaaattcttt taaggaggta aaaaatgaaa gaagtagtaa
     1981 tagcaagtgg cgtaagaaca gcggtcggaa aatttggtgg aaccctttta aacgttccag
     2041 ctgtggacct gggtgcggtg gtaattaaag aggctttaaa aagggcaaat gtaaaaccgg
     2101 aagacgtttc agaggttatt atgggaaatg ttttgcaggc aggattggga caaaatccgg
     2161 caaggcaggc tgaaataaag gcaggcatac cggtggaagt tcctgcaatg acagtgaata
     2221 tggtatgcgg aagcggctta agagctgtaa cattggctgc acaggccgtt atgctgggag
     2281 acgcagatat tgtagttgcg ggaggtatgg aaaacatgtc cagggctcct tatattttaa
     2341 atgatgcaag attcggatac agaatgaata acggacagct tgttgatgaa atggtttatg
     2401 acggacttac cgatgttttt aatcaatatc atatgggaat tactgcggaa aatcttgcag
     2461 aaaaatatgg aataagcagg gaagaacagg atgaatttgc atacaggagt caaaagctgg
     2521 cttcggaagc aatttcttcc ggaagatttg aagatgaaat tgttcccgtt atagttcccc
     2581 aaaaaaaggg agaacctatt gaattcaaag ttgacgaaca tgtgagaccg aatacaacca
     2641 tagaagcgct ggcaaagctt aaacctgcct ttaaaaaaga cggtactgta acggccggaa
     2701 acgcaagcgg tataaatgac gctgctgcgg cagttgttgt aatgtcaaag gaaaaggctg
     2761 atgagcttgg aatcaagcct cttgccacaa taaaaagctt tggatatgcc ggtgtggatc
     2821 catcaataat gggcattggt cccgtatacg caacaagaaa agcattggag aaagccaacc
     2881 tcacggtcga tgatttggac cttattgagg ccaatgaagc ttttgccgcc cagtctcttg
     2941 ctgttgcaaa agaacttaag tttaacatgg atcgggtaaa tgtaaacggc ggtgccatag
     3001 cgattggaca tccgatagga gcatcgggct gcagaatact tgtaactctt ttgtatgaaa
     3061 tgcagaaaag aaattcccac accggtctgg caactttgtg tataggcgga ggaatgggta
     3121 ttgccatggt ggttgaaaga ggtcatcacc accaccacca ttagggtctc gcggccgcca
     3181 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3241 tgctgaaagg aggaactata tccgggtaac gaattcaagc ttgatatcat tcaggacgag
     3301 cctcagactc cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc
     3361 ctttcttcgg tagaagtctt ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
     3421 gttccac
//
