LOCUS       Exported                6400 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLT_88, complete sequence.
ACCESSION   MF503104
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6400)
  AUTHORS   Tian L.
  TITLE     Elucidation of the ethanol tolerance mechanism of Clostridium 
            thermocellum by metabolome analysis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6400)
  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 6400)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6400
                     /organism="Cloning vector pLT_88"
                     /mol_type="other DNA"
                     /db_xref="taxon:2213001"
     CDS             1..1005
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     CDS             1281..2288
                     /codon_start=1
                     /gene="Gapdh"
                     /product="Gapdh"
                     /label=Gapdh
                     /note="derived from Thermoanaerobacterium saccharolyticum"
                     /protein_id="AWT04834.1"
                     /translation="MAVKVGINGFGRIGRNFFRAALKKNVDLDIVAFNDLTDAKTLAHL
                     LKYDSTFGQFEGEVIAKEDSLVVNGKEIKILKETDPAKLPWKELGVDIVIESTGRFTNK
                     EDAVKHIEAGAKKVIISAPAKNEDITIVMGVNEDKYDPNAHHVISNASCTTNCLAPFAK
                     VLHNKFGIKRGLMTTVHSYTNDQRILDLPHKDLRRARSAAMSIIPTTTGAAKAVALVLP
                     ELKGKLNGFAMRVPTPDVSVVDLVAELEKSVTVEEVNAALKEAAENELKGILGYTDEPL
                     VSMDFKGDSRSSIVDGLSTMVMEGNMVKVVSWYDNEWGYSNRVVDLAKYVADRL"
     gene            1281..2288
                     /gene="Gapdh"
                     /label=Gapdh
     CDS             2430..3080
                     /codon_start=1
                     /product="chloramphenicol resistance protein"
                     /label=chloramphenicol resistance protein
                     /protein_id="AWT04835.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     CDS             complement(3158..4018)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AWT04836.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4019..4110)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4869..5091
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5139..6089
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"