pLoxGentrc vector (V004957)

Basic Vector Information

Vector Name:
pLoxGentrc
Antibiotic Resistance:
Ampicillin
Length:
6295 bp
Type:
Expression vector
Replication origin:
ori
Source/Author:
Palmeros B, Wild J, Szybalski W, Le Borgne S, Hernandez-Chavez G, Gosset G, Valle F, Bolivar F.

pLoxGentrc vector Map

pLoxGentrc6295 bp30060090012001500180021002400270030003300360039004200450048005100540057006000f1 oriM13 fwdKS primerloxPGmRPc promoterloxP site; DNA target site for Cre site-specific recombinaserrnB T2 terminatorrrnBMCSfrom lacZlac operatortrc promoterlacIlacIq promoterlacIq 5' sequenceintergenic region from pTrc99AM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

pLoxGentrc vector Sequence

LOCUS       40924_28606        6295 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLoxGentrc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6295)
  AUTHORS   Palmeros B, Wild J, Szybalski W, Le Borgne S, Hernandez-Chavez G, 
            Gosset G, Valle F, Bolivar F.
  TITLE     A family of removable cassettes designed to obtain 
            antibiotic-resistance-free genomic modifications of Escherichia coli
            and other bacteria
  JOURNAL   Gene 247 (1-2), 255-264 (2000)
  PUBMED    10773465
REFERENCE   2  (bases 1 to 6295)
  AUTHORS   Sabido A, Martinez LM, de Anda R, Martinez A, Bolivar F, Gosset G.
  TITLE     A novel plasmid vector designed for chromosomal gene integration and
            expression: Use for developing a genetically stable Escherichia coli
            melanin production strain
  JOURNAL   Plasmid 69 (1), 16-23 (2013)
  PUBMED    22884755
REFERENCE   3  (bases 1 to 6295)
  AUTHORS   Sabido A, Gosset G.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-MAY-2011) Cell Engineering and Biocatalysis, 
            Biotechnology Institute, Av. Universidad 2001, Cuernavaca, Morelos 
            62210, Mexico
REFERENCE   4  (bases 1 to 6295)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6295)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene 247 
            (1-2), 255-264 (2000)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2013"; volume: "69"; issue: "1"; pages: "16-23"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (11-MAY-2011) Cell Engineering and Biocatalysis, Biotechnology 
            Institute, Av. Universidad 2001, Cuernavaca, Morelos 62210, Mexico"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     669..685
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     733..749
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    763..796
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(843..1373)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     regulatory      complement(1557..1563)
                     /gene="aacC1"
                     /regulatory_class="minus_10_signal"
     promoter        complement(1562..1590)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     regulatory      complement(1585..1590)
                     /gene="aacC1"
                     /regulatory_class="minus_35_signal"
     misc_feature    complement(1599..1632)
                     /note="loxP site; DNA target site for Cre site-specific 
                     recombinase"
     protein_bind    complement(1599..1632)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     terminator      complement(1716..1743)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1835..1921)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rRNA            complement(1912..2041)
                     /gene="rrnB"
                     /product="5S ribosomal RNA"
                     /label=rrnB
     misc_feature    complement(2124..2180)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      complement(2192..2195)
                     /label=from lacZ
                     /note="from lacZ"
                     /regulatory_class="ribosome_binding_site"
     protein_bind    2204..2220
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(2228..2257)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             complement(2492..3571)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3572..3649)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    complement(3650..3680)
                     /label=lacIq 5' sequence
                     /note="lacIq 5' sequence"
     misc_feature    complement(3681..3873)
                     /label=intergenic region from pTrc99A
                     /note="intergenic region from pTrc99A"
     primer_bind     complement(3888..3904)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3912..3928)
                     /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(3936..3966)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3981..4002)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4290..4878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5052..5909)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5910..6014)
                     /label=AmpR promoter

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