pATE vector (V009409)

Basic Vector Information

Vector Name:
pATE
Antibiotic Resistance:
Chloramphenicol
Length:
3828 bp
Type:
Cloning vector
Replication origin:
p15A ori
Source/Author:
Buth SA, Shneider MM, Scholl D, Leiman PG.

pATE vector Vector Map

pATE3828 bp60012001800240030003600T7 promoterlac operatorRBS6xHisTEV sitemultiple cloning siteS-TagT7 terminatorCmRcat promoterp15A oriCAP binding sitelacIlacI promoter

pATE vector Sequence

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LOCUS       40924_4994        3828 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pATE, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3828)
  AUTHORS   Buth SA, Shneider MM, Scholl D, Leiman PG.
  TITLE     Structure and Analysis of R1 and R2 Pyocin Receptor-Binding Fibers
  JOURNAL   Viruses 10 (8), E427 (2018)
  PUBMED    30110933
REFERENCE   2  (bases 1 to 3828)
  AUTHORS   Shneider M.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUL-2018) Laboratory of Molecular Engineering, IBCH 
            RAN, Miklukho-Maklaya str, Moscow 117997, Russia
REFERENCE   3  (bases 1 to 3828)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3828)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Viruses"; 
            date: "2018"; volume: "10"; issue: "8"; pages: "E427"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUL-2018) Laboratory of Molecular Engineering, IBCH RAN, 
            Miklukho-Maklaya str, Moscow 117997, Russia"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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             79..101
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             120..137
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             147..167
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     misc_feature    171..217
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             223..267
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      319..366
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(589..1245)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1246..1348)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(1874..2418)
                     /direction=LEFT
                     /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."
     protein_bind    complement(2609..2630)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2646..3725)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3726..3803)
                     /label=lacI promoter

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