pUCGM-lox vector (V002381)

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LOCUS       40924_45258        3695 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gene deletion vector pUCGM-lox, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3695)
  AUTHORS   Quenee L, Lamotte D, Polack B.
  TITLE     Combined sacB-based negative selection and cre-lox antibiotic marker
            recycling for efficient gene deletion in pseudomonas aeruginosa
  JOURNAL   BioTechniques 38 (1), 63-67 (2005)
  PUBMED    15679087
REFERENCE   2  (bases 1 to 3695)
  AUTHORS   Quenee L, Lamotte D, Polack B.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JAN-2005) GREPI, CHU, Hopital Michallon, Grenoble 
            38043, France
REFERENCE   3  (bases 1 to 3695)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3695)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "2005"; volume: "38"; issue: "1"; pages: 
            "63-67"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-JAN-2005) GREPI, CHU, Hopital Michallon, Grenoble 38043, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    492..525
                     /label=Cre recombinase binding site
                     /bound_moiety="Cre recombinase"
                     /note="loxP, Cre recombinase recognition site"
     promoter        534..562
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             751..1281
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(1328..1361)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1474..1490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1498..1514)
                     /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(1522..1552)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1567..1588)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1876..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2638..3495)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3496..3600)
                     /label=AmpR promoter