LOCUS       Exported                3268 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLT_78, complete sequence.
ACCESSION   MF503105
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3268)
  AUTHORS   Tian L.
  TITLE     Elucidation of the ethanol tolerance mechanism of Clostridium 
            thermocellum by metabolome analysis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3268)
  AUTHORS   Tian L.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-JUL-2017) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Dr, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 3268)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3268
                     /organism="Cloning vector pLT_78"
                     /mol_type="other DNA"
                     /db_xref="taxon:2212999"
     terminator      37..84
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      182..211
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      308..896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      1028..1055
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1079..1888)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AWT04837.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(1889..1979)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      1988..2018
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2083..2201
                     /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)"
     RBS             2220..2228
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             2240..2257
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2258..3268
                     /codon_start=1
                     /gene="Gapdh"
                     /product="Gapdh"
                     /label=Gapdh
                     /note="derived from Clostridium thermocellum"
                     /protein_id="AWT04838.1"
                     /translation="MAVKIGINGFGRIGRLVFRASLNNPNVEVVGINDPFIDLEYMQYM
                     LKYDTVHGQFKGEISQDNGKLVVNGRKISVYGFTDPAEIPWSECGAEYIVESTGVFTTT
                     EKASAHFKGGAKKVVISAPSADAPMFVMGVNHDKYTKDMNVVSNASCTTNCLAPLAKVI
                     HENFGIVEGLMTTVHATTATQKTVDGPSKKDWRGGRAAAGNIIPSSTGAAKAVGKVIPE
                     LNGKLTGMAFRVPTLDVSVVDLTCRLEKPATYDEIKAAVKKASENELKGILGYTEDAVV
                     SSDFIGDPRTSIFDAEAGISLNSNFVKLVAWYDNEWGYSNKVVDLIVHMASVDAK"
     gene            2258..3268
                     /gene="Gapdh"
                     /label=Gapdh