pALH015 vector (V016225)

Basic Vector Information

Vector Name:
pALH015
Antibiotic Resistance:
Ampicillin
Length:
5505 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Herneisen AL, Sidik SM, Markus BM, Drewry DH

pALH015 vector Map

pALH0155505 bp60012001800240030003600420048005400f1 oriM13 fwdT7 promoterKS primer5'UTRGCaMP6f3'UTRT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

pALH015 vector Sequence

LOCUS       62056_2970        5505 bp DNA     circular SYN 15-JUL-2020
DEFINITION  Cloning vector pALH015, complete sequence.
ACCESSION   MT345687
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5505)
  AUTHORS   Herneisen AL, Sidik SM, Markus BM, Drewry DH, Zuercher WJ, Lourido 
            S.
  TITLE     Identifying the Target of an Antiparasitic Compound in Toxoplasma 
            Using Thermal Proteome Profiling
  JOURNAL   ACS Chem. Biol. (2020) In press
  PUBMED    32597628
REFERENCE   2  (bases 1 to 5505)
  AUTHORS   Herneisen AL, Sidik SM, Markus BM, Zuercher WJ, Lourido S.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-APR-2020) Whitehead Institute, Massachusetts Institute
            of Technology, 455 Main St, Cambridge, MA 02142, USA
REFERENCE   3  (bases 1 to 5505)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Chem. 
            Biol. (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-APR-2020) Whitehead Institute, Massachusetts Institute of 
            Technology, 455 Main St, Cambridge, MA 02142, USA"
FEATURES             Location/Qualifiers
     source          1..5505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     5'UTR           698..1195
                     /label=TgTUB1 5'UTR
                     /note="TgTUB1 5'UTR"
     CDS             1196..2545
                     /label=GCaMP6f
                     /note="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
     3'UTR           2549..3291
                     /label=3'UTR of DHFR
                     /note="3'UTR of DHFR"
     promoter        complement(3317..3335)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3356..3372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3380..3396)
                     /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(3404..3434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3449..3470)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3758..4346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4520..5377)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5378..5482)
                     /label=AmpR promoter

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