LOCUS       Exported                8513 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJLO21, complete sequence.
ACCESSION   KY296113
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8513)
  AUTHORS   Lo J, Olson DG, Murphy SJ, Tian L, Hon S, Lanahan A, Guss AM, Lynd 
            LR.
  TITLE     Engineering electron metabolism to increase ethanol production in 
            Clostridium thermocellum
  JOURNAL   Metab. Eng. (2016) In press
  PUBMED    27989806
REFERENCE   2  (bases 1 to 8513)
  AUTHORS   Lo J, Olson DG, Murphy SJ, Tian L, Hon S, Lanahan A, Guss AM, Lynd 
            LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-DEC-2016) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8513)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8513
                     /organism="Cloning vector pJLO21"
                     /mol_type="other DNA"
                     /db_xref="taxon:1930519"
     CDS             complement(558..1418)
                     /codon_start=1
                     /product="Bla"
                     /EC_number="3.5.2.6"
                     /label=Bla
                     /note="beta-lactamase"
                     /protein_id="APQ46116.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1419..1523)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3191..3841
                     /codon_start=1
                     /product="Cat"
                     /EC_number="2.3.1.28"
                     /label=Cat
                     /note="chloramphenicol transferase"
                     /protein_id="APQ46117.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     CDS             3859..4404
                     /codon_start=1
                     /product="Hpt"
                     /EC_number="2.4.2.8"
                     /label=Hpt
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="APQ46118.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     rep_origin      complement(5559..6147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6234..6812)
                     /codon_start=1
                     /product="Tdk"
                     /EC_number="2.7.1.21"
                     /label=Tdk
                     /note="thymidine kinase"
                     /protein_id="APQ46119.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     CDS             complement(7509..8513)
                     /codon_start=1
                     /product="RepB"
                     /EC_number="3.6.4.12"
                     /label=RepB
                     /note="replication protein"
                     /protein_id="APQ46120.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"