pYS2 vector (V001868)

Basic Vector Information

Vector Name:
pYS2
Antibiotic Resistance:
Ampicillin
Length:
5315 bp
Type:
Gene deletion vector
Replication origin:
ori
Source/Author:
Shenkerman Y, Elharar Y, Vishkautzan M, Gur E.

pYS2 vector Map

pYS25315 bp6001200180024003000360042004800CAP binding sitelac promoterlac operatorM13 revloxPmgfp5Hygromycin-B 7''-O-kinaseloxP site for cre recombinaseAmpR promoterAmpRori

pYS2 vector Sequence

LOCUS       V001868                 5315 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V001868
VERSION     V001868
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5315)
  AUTHORS   Shenkerman Y, Elharar Y, Vishkautzan M, Gur E.
  TITLE     Efficient and simple generation of unmarked gene deletions in
            Mycobacterium smegmatis
  JOURNAL   Gene 533 (1), 374-378 (2014)
   PUBMED   24100088
REFERENCE   2  (bases 1 to 5315)
  AUTHORS   Shenkerman Y, Elharar Y, Vishkautzan M, Gur E.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-AUG-2013) Life Sciences, Ben-Gurion University of the
            Negev, Rager Blvd, Beer-Sheva 84105, Israel
REFERENCE   3  (bases 1 to 5315)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5315)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
            date: "2014"; volume: "533"; issue: "1"; pages: "374-378"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-AUG-2013) Life Sciences, Ben-Gurion University of the Negev,
            Rager Blvd, Beer-Sheva 84105, Israel"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    616..649
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1008..1721
                     /label="mgfp5"
                     /note="GFP with folding enhancement mutations"
     CDS             1836..2831
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     misc_recomb     3059..3092
                     /label="loxP site for cre recombinase"
                     /note="loxP site for cre recombinase"
     protein_bind    complement(3059..3092)
                     /label="loxP"
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     promoter        3409..3513
                     /label="AmpR promoter"
     CDS             3514..4371
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4545..5133
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"

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