pJY16 vector (V015364)

LOCUS       V015364                 9728 bp    DNA     circular SYN 16-FEB-2021
DEFINITION  Exported.
ACCESSION   V015364
VERSION     V015364
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9728)
  AUTHORS   Yayo J, Kuil T, Olson DG, Lynd LR, Holwerda EK, van Maris AJA.
  TITLE     Laboratory evolution and reverse engineering of Clostridium
            thermocellum for growth on glucose and fructose
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9728)
  AUTHORS   Yayo J, Kuil T, Olson DG, Lynd LR, Holwerda EK, van Maris AJA.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-DEC-2020) Industrial Biotechnology, KTH Royal
            Institute of Technology, Roslagstullsbacken 21, Stockholm 11421,
            Sweden
REFERENCE   3  (bases 1 to 9728)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (09-DEC-2020) Industrial Biotechnology, KTH Royal Institute of
            Technology, Roslagstullsbacken 21, Stockholm 11421, Sweden"
FEATURES             Location/Qualifiers
     source          1..9728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..455
                     /label="C. thermocellum origin of replication"
                     /note="C. thermocellum origin of replication"
     CDS             456..1457
                     /label="repB"
                     /note="RepB replication protein"
     regulatory      1536..2156
                     /label="C. thermocellum cbp promoter"
                     /note="C. thermocellum cbp promoter"
                     /regulatory_class="promoter"
     gene            2157..2735
                     /gene="tdk"
                     /label="tdk"
     CDS             2157..2735
                     /codon_start=1
                     /transl_table=11
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /protein_id="QRQ89514.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     rep_origin      complement(3002..3547)
                     /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."
     terminator      complement(3597..3785)
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     misc_feature    complement(3821..4409)
                     /label="upstream homology recombination 5' flank"
                     /note="upstream homology recombination 5' flank"
     gene            complement(4594..5139)
                     /gene="hpt"
                     /label="hpt"
     CDS             complement(4594..5139)
                     /codon_start=1
                     /transl_table=11
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /protein_id="QRQ89515.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     CDS             complement(5160..5807)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     regulatory      complement(5808..6382)
                     /label="C. thermocellum gapDH promoter"
                     /note="C. thermocellum gapDH promoter"
                     /regulatory_class="promoter"
     misc_feature    complement(6386..7007)
                     /label="downstream homology recombination 3' flank"
                     /note="downstream homology recombination 3' flank"
     gene            7014..7979
                     /gene="cbpA"
                     /label="cbpA"
                     /note="gene target with single-point mutation c.443G>T
                     resulting in G148V"
     CDS             7014..7979
                     /codon_start=1
                     /transl_table=11
                     /gene="cbpA"
                     /product="carbohydrate binding protein A"
                     /label="cbpA"
                     /note="gene target CDS with single-point mutation c.443G>T
                     resulting in G148V"
                     /protein_id="QRQ89517.1"
                     /translation="MKRFLVLLLAISLIFTFTACNSGSGSGNGQAKGYVGDPSDEYYMV
                     TFLSGIDYWKYCFEGFEDAAKAIGVTAKYTGQTDTDVSGQVAVLEQVIAQKPKGIAVTA
                     VNSTALADTINSAIEQGISVVCFDSDSPTSNRSAYLGTGNYAAVQKAAEFLVPLVNYKG
                     KIAVLYTVGAENSESRVQGFEDWCKQNAPEVSLVKVNDAGDTTVAADNLAAALQANDDI
                     VGVFCVDGVAGTAGPTAVAESKKDLRVLAFDVDVTVLDKVKSGEIDGTVAQGMYNMGYW
                     SLMMLYTEANGLSSKALPGNLDTGVVIVTKDNVDEYYPKK"
     misc_feature    complement(7980..8568)
                     /label="upstream homology recombination 5' flank"
                     /note="upstream homology recombination 5' flank"
     promoter        8661..8765
                     /label="AmpR promoter"
     CDS             8766..9623
                     /label="AmpR"
                     /note="beta-lactamase"