pTac-VvSTS-At4CL1m vector (V002826)

Basic Vector Information

Vector Name:
pTac-VvSTS-At4CL1m
Antibiotic Resistance:
Kanamycin
Length:
7315 bp
Type:
Cloning vector
Replication origin:
p15A ori
Source/Author:
Kim B, Binkley R, Kim HU, Lee SY.
Promoter:
tac

pTac-VvSTS-At4CL1m vector Vector Map

pTac-VvSTS-At4CL1m7315 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200Stilbene synthase 2 from Vitis vinifera. Accession#: P51070rrnB T1 terminatorrrnB T2 terminatorAmpR promoterM13 fwdtac promoterlac operator4CL1rrnB T1 terminatorrrnB T2 terminatorKanRp15A oriTac promoterlac operator

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pTac-VvSTS-At4CL1m vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_42484        7315 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-VvSTS-At4CL1m, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7315)
  AUTHORS   Kim B, Binkley R, Kim HU, Lee SY.
  TITLE     Metabolic engineering of Escherichia coli for the enhanced 
            production of l-tyrosine
  JOURNAL   Biotechnol. Bioeng. (2018) In press
  PUBMED    30019750
REFERENCE   2  (bases 1 to 7315)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUN-2018) Dept. Chemical and Biomolecular Engineering,
            Korea Advanced Institute of Science and Technology, Daehak-ro 291, 
            Daejeon 34141, South Korea
REFERENCE   3  (bases 1 to 7315)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7315)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Bioeng. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JUN-2018) Dept. Chemical and Biomolecular Engineering, Korea 
            Advanced Institute of Science and Technology, Daehak-ro 291, Daejeon
            34141, South Korea"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..1183
                     /note="Stilbene synthase 2 from Vitis vinifera. Accession#:
                     P51070"
     terminator      1435..1521
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1613..1640
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1660..1751
                     /label=AmpR promoter
     primer_bind     complement(2064..2080)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2325..2353
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    2361..2377
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2399..4081
                     /gene="4CL1"
                     /label=4CL1
                     /note="4-coumarate--CoA ligase 1 from Arabidopsis thaliana.
                     Accession#: Q42524"
     terminator      complement(4336..4379)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4511..4538
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             4723..5535
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      6067..6612
                     /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."
     regulatory      7249..7277
                     /label=Tac promoter
                     /note="Tac promoter"
                     /regulatory_class="promoter"
     promoter        7249..7277
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7285..7301
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

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