LOCUS       Exported                9005 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pZJ08, complete sequence.
ACCESSION   KU682276
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9005)
  AUTHORS   Zhou J, Olson DG.
  TITLE     Conversion of phosphoenolpyruvate to pyruvate in Clostridium 
            thermocellum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9005)
  AUTHORS   Zhou J, Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2016) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 9005)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9005
                     /organism="Cloning vector pZJ08"
                     /mol_type="other DNA"
                     /db_xref="taxon:1889897"
     CDS             complement(1..861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="Bla; ampicillin resistance protein; provides 
                     antibiotic resistance to ampicillin and carbenicillin"
                     /protein_id="AOC59183.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1..861)
                     /gene="bla"
                     /label=bla
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2986..3636
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="Cat; provides antibiotic resistance to 
                     thiamphenicol"
                     /protein_id="AOC59184.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            2986..3636
                     /gene="cat"
                     /label=cat
     CDS             3654..4199
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label=hpt
                     /note="Hpt; counterselection with 8AZH"
                     /protein_id="AOC59185.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            3654..4199
                     /gene="hpt"
                     /label=hpt
     rep_origin      5435..6108
                     /label=E. coli origin of replication
                     /note="E. coli origin of replication"
     rep_origin      complement(5494..6082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6169..6747)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label=tdk
                     /note="Tdk; counterselection with FUDR"
                     /protein_id="AOC59186.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(6169..6747)
                     /gene="tdk"
                     /label=tdk
     CDS             complement(7444..8448)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein"
                     /label=repB
                     /protein_id="AOC59187.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(7444..8448)
                     /gene="repB"
                     /label=repB
     rep_origin      complement(8449..8903)
                     /direction=LEFT
                     /label=C. thermocellum replication origin
                     /note="C. thermocellum replication origin"