Basic Vector Information
- Vector Name:
- pET-Sac_rppA
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6341 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Kim B, Binkley R, Kim HU, Lee SY.
pET-Sac_rppA vector Map
pET-Sac_rppA vector Sequence
LOCUS 40924_18231 6341 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pET-Sac_rppA, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6341) 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 6341) AUTHORS Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY. TITLE Direct Submission JOURNAL Submitted (14-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 6341) TITLE Direct Submission REFERENCE 4 (bases 1 to 6341) 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 (14-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..6341 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 12..467 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(563..1375) /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 1497..2085 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(2271..2413) /label=bom /note="basis of mobility region from pBR322" CDS complement(2518..2706) /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" protein_bind complement(3481..3502) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(3518..4597) /codon_start=1 /label=lacI /note="lac repressor" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(4598..4675) /label=lacI promoter promoter 4988..5006 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5007..5031 /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)." RBS 5046..5068 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 5076..6143 /codon_start=1 /gene="rppA" /product="1,3,6,8-tetrahydroxynaphthalene synthase" /label=rppA /note="derived from Streptomyces aculeolatus" /protein_id="AXN69953.1" /translation="MPRLCKPAVSAPEYTITMEETLEFAKQAHAGKPQLPLALRLIRNT GVLKRHIVQPIEKTLGHPGLTERNLIYEAESKKMCPPVIEEALQNADMTARDIDAIIYV SCTGFLMPSLTAWLINKMGFRSDTRQIPIAQLGCAAGGAAVNRAHDFCLAHPGSNVLIV ACELCSLCYQPTADDIGSLLSDGLFGDAVAAAVVRGNGGVGIEVERNASYLIPNTEEWI SYSVRDTGFHFQLDRRVPGTMEPLAPVLREFAKDHSWDAGKLDFYIVHAGGPRILDDLA RFLDVDRQVFRHSWSTLTEYGNIASAVVFDAARRLFEEGSAKPDATGMIAGFGPGITAE MALGTWGTDGTGTSN" gene 5076..6143 /gene="rppA" /label=rppA CDS 6185..6202 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" terminator 6269..6316 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase"
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