Basic Vector Information
pTac-Sc4CL 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-Sc4CL vector Sequence
LOCUS 40924_42469 5305 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pTac-Sc4CL, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5305) 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 5305) 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 5305) TITLE Direct Submission REFERENCE 4 (bases 1 to 5305) 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..5305 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 8..1576 /codon_start=1 /gene="Sc4CL" /product="4-coumarate:CoA ligase" /label=Sc4CL /note="derived from Streptomyces coelicolor" /protein_id="AXN70032.1" /translation="MFRSEYADVPPVDLPIHDAVLGGAAAFGSTPALIDGTDGTTLTYE QVDRFHRRVAAALAETGVRKGDVLALHSPNTVAFPLAFYAATRAGASVTTVHPLATAEE FAKQLKDSAARWIVTVSPLLSTARRAAELAGGVQEILVCDSAPGHRSLVDMLASTAPEP SVAIDPAEDVAALPYSSGTTGTPKGVMLTHRQIATNLAQLEPSMPSAPGDRVLAVLPFF HIYGLTALMNAPLRLGATVVVLPRFDLEQFLAAIQNHRITSLYVAPPIVLALAKHPLVA DYDLSSLRYIVSAAAPLDARLAAACSQRLGLPPVGQAYGMTELSPGTHVVPLDAMADAP PGTVGRLIAGTEMRIVSLTDPGTDLPAGESGEILIRGPQIMKGYLGRPDATAAMIDEEG WLHTGDVGHVDADGWLFVVDRVKELIKYKGFQVAPAELEAHLLTHPGVADAAVVGAYDD DGNEVPHAFVVRQPAAPGLAESEIMMYVAERVAPYKRVRRVTFVDAVPRAASGKILRRQ LREPR" gene 8..1576 /gene="Sc4CL" /label=Sc4CL 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 /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" 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 5239..5267 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 5275..5291 /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)."
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