LOCUS       Exported                7118 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLT_110, complete sequence.
ACCESSION   MF818012
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7118)
  AUTHORS   Tian L.
  TITLE     Methods for metabolic engineering of Thermoanaerobacterium 
            saccharolyticum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7118)
  AUTHORS   Tian L.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-SEP-2017) MCB, Dartmouth College, 14 Engineering 
            Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 7118)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7118
                     /organism="Cloning vector pLT_110"
                     /mol_type="other DNA"
                     /db_xref="taxon:2283382"
     rep_origin      1..828
                     /label=p15A
                     /note="p15A"
     rep_origin      complement(256..801)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             1336..3957
                     /codon_start=1
                     /product="AdhE"
                     /EC_number="1.1.1.2"
                     /label=AdhE
                     /note="bifunctional iron-containing alcohol dehydrogenase 
                     and aldehyde dehydrogenase; derived from Clostridium 
                     thermocellum"
                     /protein_id="AXJ20730.1"
                     /translation="MTKIANKYEVIDNVEKLEKALKRLREAQSVYATYTQEQVDKIFFE
                     AAMAANKMRIPLAKMAVEETGMGVVEDKVIKNHYASEYIYNAYKNTKTCGVIEEDPAFG
                     IKKIAEPLGVIAAVIPTTNPTSTAIFKTLIALKTRNAIIISPHPRAKNSTIEAAKIVLE
                     AAVKAGAPEGIIGWIDVPSLELTNLVMREADVILATGGPGLVKAAYSSGKPAIGVGAGN
                     TPAIIDDSADIVLAVNSIIHSKTFDNGMICASEQSVIVLDGVYKEVKKEFEKRGCYFLN
                     EDETEKVRKTIIINGALNAKIVGQKAHTIANLAGFEVPETTKILIGEVTSVDISEEFAH
                     EKLCPVLAMYRAKDFDDALDKAERLVADGGFGHTSSLYIDTVTQKEKLQKFSERMKTCR
                     ILVNTPSSQGGIGDLYNFKLAPSLTLGCGSWGGNSVSDNVGVKHLLNIKTVAERRENML
                     WFRTPEKIYIKRGCLPVALDELKNVMGKKKAFIVTDNFLYNNGYTKPITDKLDEMGIVH
                     KTFFDVSPDPSLASAKAGAAEMLAFQPDTIIAVGGGSAMDAAKIMWVMYEHPEVDFMDM
                     AMRFMDIRKRVYTFPKMGQKAYFIAIPTSAGTGSEVTPFAVITDEKTGIKYPLADYELL
                     PDMAIVDADMMMNAPKGLTAASGIDALTHALEAYVSMLATDYTDSLALRAIKMIFEYLP
                     RAYENGASDPVAREKMANAATIAGMAFANAFLGVCHSMAHKLGAFYHLPHGVANALMIN
                     EVIRFNSSEAPTKMGTFPQYDHPRTLERYAEIADYIGLKGKNNEEKVENLIKAIDELKE
                     KVGIRKTIKDYDIDEKEFLDRLDEMVEQAFDDQCTGTNPRYPLMNEIRQMYLNAYYGGA
                     KK"
     regulatory      4533..5063
                     /regulatory_class="promoter"
     CDS             5064..5825
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /note="HTK"
                     /protein_id="AXJ20731.1"
                     /translation="MKGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVLSTEGVEFSYEWTTGEWKAEVNFYSEEILLDYASRVEPDWPLTHGRFF
                     SILPIYDPGGYFEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQPDLPPGYVQLCQLVMSGQLSDPEK
                     LLESLENFWNGVQEWAERHGYIVDVSKRIPF"
     CDS             5849..6430
                     /codon_start=1
                     /product="thymidine kinase"
                     /EC_number="2.7.1.21"
                     /label=thymidine kinase
                     /note="tdk"
                     /protein_id="AXJ20732.1"
                     /translation="MIYGSLDHGFIEAIVGPMFSGKSEELIRRIKRAQIAKQKVQVFKP
                     AIDDRYSIDKVVSHNGTNINAISVVKASEIIELLEEDTEVIAIDEIQFFDHSIVDVVRE
                     IADLGKRVICAGLDMDFRGEPFGPTPDIMAIAESVDKLTAICVKCGNPATRTQRLINGK
                     PAKYDDPIILVGAHETYEARCRKCHEVPRT"