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pTrc-At4CL1m-AtCHI-PhCHS vector (V002557#)

Basic Vector Information

Basic Vector Information
Vector Name pTrc-At4CL1m-AtCHI-PhCHS Antibiotic Resistance Ampicillin
Length 7827 bp Type Cloning vector
Source Kim B, Binkley R, Kim HU, Lee SY.

pTrc-At4CL1m-AtCHI-PhCHS vector Vector Map

Trc promoter 4CL1m CHI CHS rrnBT1T2 AmpR promoter bla pBR322 bom lacIq promoter

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 ℃.

pTrc-At4CL1m-AtCHI-PhCHS vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7827 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTrc-At4CL1m-AtCHI-PhCHS, complete sequence.
ACCESSION   MH488946
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7827)
  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 7827)
  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 7827)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7827
                     /organism="Cloning vector pTrc-At4CL1m-AtCHI-PhCHS"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:2301172"
     regulatory      193..262
                     /regulatory_class="promoter"
                     /note="Trc promoter"
     promoter        193..222
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     CDS             267..1952
                     /codon_start=1
                     /gene="4CL1m"
                     /product="4-coumarate:CoA ligase 1 (4AT mutant)"
                     /label=4CL1m
                     /note="I250L/N404K/I461V; derived from Arabidopsis 
                     thaliana"
                     /protein_id="AXN70059.1"
                     /translation="MAPQEQAVSQVMEKQSNNNNSDVIFRSKLPDIYIPNHLSLHDYIF
                     QNISEFATKPCLINGPTGHVYTYSDVHVISRQIAANFHKLGVNQNDVVMLLLPNCPEFV
                     LSFLAASFRGATATAANPFFTPAEIAKQAKASNTKLIITEARYVDKIKPLQNDDGVVIV
                     CIDDNESVPIPEGCLRFTELTQSTTEASEVIDSVEISPDDVVALPYSSGTTGLPKGVML
                     THKGLVTSVAQQVDGENPNLYFHSDDVLLCVLPMFHIYALNSIMLCGLRVGAAILIMPK
                     FEINLLLELIQRCKVTVAPMVPPIVLAIAKSSETEKYDLSSIRVVKSGAAPLGKELEDA
                     VNAKFPNAKLGQGYGMTEAGPVLAMSLGFAKEPFPVKSGACGTVVRNAEMKIVDPDTGD
                     SLSRKQPGEICIRGHQIMKGYLNNPAATAETIDKDGWLHTGDIGLIDDDDELFIVDRLK
                     ELVKYKGFQVAPAELEALLIGHPDITDVAVVAMKEEAAGEVPVAFVVKSKDSELSEDDV
                     KQFVSKQVVFYKRINKVFFTESIPKAPSGKILRKDLRAKLANGL"
     gene            267..1952
                     /gene="4CL1m"
                     /label=4CL1m
     CDS             1978..2718
                     /codon_start=1
                     /gene="CHI"
                     /product="chalcone isomerase"
                     /label=CHI
                     /note="derived from Arabidopsis thaliana"
                     /protein_id="AXN70060.1"
                     /translation="MSSSNACASPSPFPAVTKLHVDSVTFVPSVKSPASSNPLFLGGAG
                     VRGLDIQGKFVIFTVIGVYLEGNAVPSLSVKWKGKTTEELTESIPFFREIVTGAFEKFI
                     KVTMKLPLTGQQYSEKVTENCVAIWKQLGLYTDCEAKAVEKFLEIFKEETFPPGSSILF
                     ALSPTGSLTVAFSKDDSIPETGIAVIENKLLAEAVLESIIGKNGVSPGTRLSVAERLSQ
                     LMMKNKDEKEVSDHSVEEKLAKEN"
     gene            1978..2718
                     /gene="CHI"
                     /label=CHI
     CDS             2778..3947
                     /codon_start=1
                     /gene="CHS"
                     /product="chalcone synthase"
                     /label=CHS
                     /note="derived from Petunia x hybrida"
                     /protein_id="AXN70061.1"
                     /translation="MVTVEEYRKAQRAEGPATVMAIGTATPTNCVDQSTYPDYYFRITN
                     SEHKTDLKEKFKRMCEKSMIKKRYMHLTEEILKENPSMCEYMAPSLDARQDIVVVEVPK
                     LGKEAAQKAIKEWGQPKSKITHLVFCTTSGVDMPGCDYQLTKLLGLRPSVKRLMMYQQG
                     CFAGGTVLRLAKDLAENNKGARVLVVCSEITAVTFRGPNDTHLDSLVGQALFGDGAGAI
                     IIGSDPIPGVERPLFELVSAAQTLLPDSHGAIDGHLREVGLTFHLLKDVPGLISKNIEK
                     SLEEAFRPLSISDWNSLFWIAHPGGPAILDQVEIKLGLKPEKLKATRNVLSNYGNMSSA
                     CVLFILDEMRKASAKEGLGTTGEGLEWGVLFGFGPGLTVETVVLHSVAT"
     gene            2778..3947
                     /gene="CHS"
                     /label=CHS
     regulatory      4034..4397
                     /regulatory_class="terminator"
                     /note="rrnBT1T2"
     terminator      4180..4266
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4358..4385
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4405..4496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4497..5357
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AXN70062.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4497..5357
                     /gene="bla"
                     /label=bla
     rep_origin      5512..6131
                     /label=pBR322
                     /note="pBR322"
     rep_origin      5528..6116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    6302..6442
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        6628..6705
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6697..7788
                     /codon_start=1
                     /gene="lacIq"
                     /product="lac repressor protein"
                     /label=lacIq
                     /protein_id="AXN70063.1"
                     /translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
                     SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
                     SSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
                     SPRALADSLMQLARQVSRLESGQ"
     gene            6697..7788
                     /gene="lacIq"
                     /label=lacIq
     CDS             6706..7788
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"

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