Basic Vector Information
pTac-VvSTS-Sc4CLm vector Vector Map
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-Sc4CLm vector Sequence
LOCUS 40924_42499 7194 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTac-VvSTS-Sc4CLm, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7194)
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 7194)
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 7194)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7194)
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..7194
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 8..1576
/codon_start=1
/gene="Sc4CLm"
/product="4-coumarate:CoA ligase mutant"
/label=Sc4CLm
/note="A294G; derived from Streptomyces coelicolor"
/protein_id="AXN70047.1"
/translation="MFRSEYADVPPVDLPIHDAVLGGAAAFGSTPALIDGTDGTTLTYE
QVDRFHRRVAAALAETGVRKGDVLALHSPNTVAFPLAFYAATRAGASVTTVHPLATAEE
FAKQLKDSAARWIVTVSPLLSTARRAAELAGGVQEILVCDSAPGHRSLVDMLASTAPEP
SVAIDPAEDVAALPYSSGTTGTPKGVMLTHRQIATNLAQLEPSMPSAPGDRVLAVLPFF
HIYGLTALMNAPLRLGATVVVLPRFDLEQFLAAIQNHRITSLYVAPPIVLALAKHPLVA
DYDLSSLRYIVSGAAPLDARLAAACSQRLGLPPVGQAYGMTELSPGTHVVPLDAMADAP
PGTVGRLIAGTEMRIVSLTDPGTDLPAGESGEILIRGPQIMKGYLGRPDATAAMIDEEG
WLHTGDVGHVDADGWLFVVDRVKELIKYKGFQVAPAELEAHLLTHPGVADAAVVGAYDD
DGNEVPHAFVVRQPAAPGLAESEIMMYVAERVAPYKRVRRVTFVDAVPRAASGKILRRQ
LREPR"
gene 8..1576
/gene="Sc4CLm"
/label=Sc4CLm
terminator 1825..1911
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2003..2030
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 2050..2141
/label=AmpR promoter
primer_bind complement(2454..2470)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 2713..3525
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 4057..4602
/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."
promoter 5037..5065
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 5073..5089
/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 5111..6286
/note="Stilbene synthase 2 from Vitis vinifera. Accession#:
P51070"
terminator complement(6541..6584)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 6716..6743
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
regulatory 7128..7156
/label=Tac promoter
/note="Tac promoter"
/regulatory_class="promoter"
promoter 7128..7156
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 7164..7180
/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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