LOCUS       40924_28811        3268 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLT_78, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3268)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-JUL-2017) Thayer School of Engineering, Dartmouth College, 14 
            Engineering Dr, Hanover, NH 03755, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1028..1055)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1082..1888)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /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
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /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
ORIGIN
        1 ggttagggtc tcgcggccgc caccgctgag caataactag cataacccct tggggcctct
       61 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccgggta acgaattcaa
      121 gcttgatatc attcaggacg agcctcagac tccagcgtaa ctggactgca atcaactcac
      181 tggctcacct tcacgggtgg gcctttcttc ggtagaagtc ttctcatgac caaaatccct
      241 taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      301 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      361 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      421 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttagccc
      481 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      541 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      601 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      661 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      721 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      781 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      841 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      901 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      961 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     1021 ccgctcgcgc agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatt
     1081 agaaaaactc atcgagcatc aaatgaaatt gcaatttatt catatcagga ttatcaatac
     1141 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     1201 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     1261 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     1321 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     1381 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     1441 cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgagt
     1501 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     1561 cttctaatac ctggaacgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat
     1621 caggagtacg gataaaatgc ttgatggtcg gaagtggcat aaattccgtc agccagttta
     1681 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     1741 actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat
     1801 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     1861 tcgacgtttc ccgttgaata tggctcatag ctcctgaaaa tctcgataac tcaaaaaata
     1921 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     1981 gaagacggtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat
     2041 gatttaaatg gtcagtattg agcgatatct agagaattcg tccaccacaa ttcagcaaat
     2101 tgtgaacatc atcacgttca tctttccctg gttgccaatg gcccattttc ctgtcagtaa
     2161 cgagaaggtc gcgaattcag gcgcttttta gactggtcgt agagaccatg aaattctttt
     2221 aaggaggtaa aaaatgaaac atcaccacca ccaccatatg gcagtaaaaa ttggtatcaa
     2281 cggttttgga cgtatcggtc gtcttgtgtt cagggccagt ctcaacaacc cgaacgttga
     2341 ggttgtaggt ataaacgacc catttattga ccttgaatac atgcaatata tgttaaagta
     2401 tgatacagta catggtcagt tcaaaggtga aatttcacaa gacaacggaa aactcgttgt
     2461 aaacggcaga aaaataagtg tttatggttt cactgatcct gctgagattc cgtggagcga
     2521 atgcggtgcg gaatacatcg ttgaatcaac gggtgtattc acaactactg aaaaggcttc
     2581 agctcacttc aagggcggtg caaagaaggt agttatcagt gctccttcgg cagatgctcc
     2641 gatgtttgtt atgggtgtaa accatgataa atacacaaag gatatgaacg ttgtatctaa
     2701 cgcttcatgt acaacaaact gccttgctcc tctggctaaa gttatacatg aaaacttcgg
     2761 aatcgtagaa ggcttgatga ctactgtaca cgctacaact gcaactcaaa agactgttga
     2821 cggtccttca aagaaagact ggagaggcgg acgtgcagca gcaggcaaca tcattccttc
     2881 atcaacagga gctgcaaagg cagttggaaa ggttatccct gaattgaacg gtaagttgac
     2941 aggtatggct ttcagagttc cgactcttga cgtatctgtt gttgacttga cttgccgtct
     3001 tgaaaaaccg gctacttatg atgaaatcaa agccgctgtt aagaaagctt cagaaaatga
     3061 acttaagggt attttgggat acactgagga tgcggttgtt tcttcagact tcatcggcga
     3121 tccccgcact tcaattttcg atgctgaagc aggtatttct ctcaacagca actttgtaaa
     3181 acttgttgcc tggtacgaca atgaatgggg ttattcaaac aaagttgttg atttgatagt
     3241 tcatatggct tcggttgatg caaaataa
//
