pLL1228 vector (V015730)

Basic Vector Information

Vector Name:
pLL1228
Antibiotic Resistance:
Ampicillin
Length:
8469 bp
Type:
Cloning vector
Replication origin:
p15A ori
Source/Author:
Holwerda EK, Zhou J, Hon S, Stevenson D

pLL1228 vector Map

pLL12288469 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400p15A oritdkC. thermocellum cbp promoterrepBC. thermocellum origin of replicationAmpRAmpR promoterupstream homologous recombination flankdownstream homologous recombination flankC. thermocellum gapdh promoterChloramphenicol acetyltransferasehptgene target internal homologous recombination regionCYC1 terminator

pLL1228 vector Sequence

LOCUS       V015730                 8469 bp    DNA     circular SYN 05-OCT-2020
DEFINITION  Exported.
ACCESSION   V015730
VERSION     V015730
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8469)
  AUTHORS   Holwerda EK, Zhou J, Hon S, Stevenson D, Amador-Noguez D, Lynd LR,
            van Dijken JP.
  TITLE     Metabolic fluxes of nitrogen and pyrophosphate in chemostat cultures
            of Clostridium thermocellum and Thermoanaerobacterium
            saccharolyticum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8469)
  AUTHORS   Hon S.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-APR-2020) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8469)
  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            "Unpublished"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
            (28-APR-2020) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, USA"
FEATURES             Location/Qualifiers
     source          1..8469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..573
                     /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."
     gene            complement(840..1418)
                     /gene="tdk"
                     /label="tdk"
     CDS             complement(840..1418)
                     /codon_start=1
                     /transl_table=11
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /protein_id="QOE54900.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     regulatory      complement(1419..2039)
                     /label="C. thermocellum cbp promoter"
                     /note="C. thermocellum cbp promoter"
                     /regulatory_class="promoter"
     CDS             complement(2118..3119)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(3120..3574)
                     /direction=LEFT
                     /label="C. thermocellum origin of replication"
                     /note="C. thermocellum origin of replication"
     CDS             complement(3680..4537)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(4538..4642)
                     /label="AmpR promoter"
     misc_feature    4732..5244
                     /label="upstream homologous recombination flank"
                     /note="upstream homologous recombination flank"
     misc_feature    5245..5736
                     /label="downstream homologous recombination flank"
                     /note="downstream homologous recombination flank"
     regulatory      5740..6314
                     /label="C. thermocellum gapdh promoter"
                     /note="C. thermocellum gapdh promoter"
                     /regulatory_class="promoter"
     CDS             6315..6962
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     gene            6983..7528
                     /gene="hpt"
                     /label="hpt"
     CDS             6983..7528
                     /codon_start=1
                     /transl_table=11
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /protein_id="QOE54904.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     misc_feature    7713..8226
                     /note="gene target internal homologous recombination
                     region"
     terminator      8259..8447
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"

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