Basic Vector Information
- Vector Name:
- pET28AMP_MaIE
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5377 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Skowron PM, Zylicz-Stachula A.
pET28AMP_MaIE vector Map
pET28AMP_MaIE vector Sequence
LOCUS 62056_10155 5377 bp DNA circular SYN 02-OCT-2019 DEFINITION Cloning vector pET28AMP_MaIE, complete sequence. ACCESSION MK606522 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5377) AUTHORS Skowron PM, Zylicz-Stachula A. TITLE Direct Submission JOURNAL Submitted (07-MAR-2019) University of Gdansk, Faculty of Chemistry, Wita Stwosza 63, Gdansk, Pomorskie 80-308, Poland REFERENCE 2 (bases 1 to 5377) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted (07-MAR-2019) University of Gdansk, Faculty of Chemistry, Wita Stwosza 63, Gdansk, Pomorskie 80-308, Poland" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..5377 /mol_type="other DNA" /organism="synthetic DNA construct" terminator complement(26..73) /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(140..157) /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" CDS complement(206..223) /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" RBS complement(314..336) /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind complement(351..375) /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)." promoter complement(376..394) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 703..780 /label=lacI promoter CDS 781..1860 /codon_start=1 /label=lacI /note="lac repressor" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" protein_bind 1876..1897 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS 2672..2860 /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" misc_feature 2965..3107 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(3293..3881) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS 4003..4815 /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin complement(4911..5366) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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