pJL2 vector (V005316)

LOCUS       40924_26196       10360 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gene deletion vector pJL2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10360)
  AUTHORS   Olson DG, Tripathi SA, Giannone RJ, Lo J, Caiazza NC, Hogsett DA, 
            Hettich RL, Guss AM, Dubrovsky G, Lynd LR.
  TITLE     Deletion of the Cel48S cellulase from Clostridium thermocellum
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 107 (41), 17727-17732 (2010)
  PUBMED    20837514
REFERENCE   2  (bases 1 to 10360)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-AUG-2010) Thayer School of Engineering, Dartmouth 
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 10360)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10360)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2010"; volume: "107"; issue: "41"; pages:
            "17727-17732"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-AUG-2010) Thayer School of Engineering, Dartmouth College, 8000 
            Cummings, Hanover, NH 03755, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      27..274
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(522..1110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      1182..1526
                     /note="pfk; derived from Clostridium thermocellum strain
                     DSM 1313"
                     /regulatory_class="promoter"
     RBS             1512..1520
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1527..2471
                     /codon_start=1
                     /product="pyrF"
                     /label=pyrF
                     /note="5FOA negative-selection in Clostridium thermocellum"
                     /protein_id="ADW95859.1"
                     /translation="MFIDTLIEKIREKDNPSVVGLDPKIEYVPSFIKEDMYKKYGKNLK
                     AVAEAILLFNKYIIDAVYDIVPAVKPQLAYYEMYGLEGMRVFYETCKYAKEKGLLVIAD
                     GKRNDIGSTAQCYSAAYLGKTDIDEGISEAVFDVDALTVNPYLGVDGIKPFIDDCVKYN
                     KGIFVLVKTSNKSSGEIQDILTQEGRSIYEIVAEYVESWGENKKGKYGYSCVGAVVGAT
                     YPNLAKILRKILKNSYILVPGYGAQGGTARDVAHCFNYDGLGAIVNASRSIMCAYKSEQ
                     WKNVYSEEKFYEASRAEAIRMRDDINSALRDRK"
     CDS             complement(2574..3431)
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             complement(3530..4330)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(4331..4551)
                     /label=URA3 promoter
     misc_feature    4579..5082
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     CDS             complement(5314..6315)
                     /label=repB
                     /note="RepB replication protein"
     misc_feature    complement(6784..7783)
                     /note="cel48Y 3' flanking region; derived from Clostridium 
                     thermocellum strain DSM 1313"
     regulatory      7784..8431
                     /note="gapDH; derived from Clostridium thermocellum strain
                     DSM 1313"
                     /regulatory_class="promoter"
     CDS             8432..9223
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    complement(9227..10281)
                     /note="cel48Y 5' flanking region; derived from Clostridium 
                     thermocellum strain DSM 1313"