pAW068 vector (V009387)

Basic Vector Information

Vector Name:
pAW068
Antibiotic Resistance:
Chloramphenicol
Length:
9406 bp
Type:
Transposon delivery vector
Replication origin:
ori
Source/Author:
Wilson AC, Perego M, Hoch JA.

pAW068 vector Map

pAW0689406 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200C9 transposaseT7 promoterCAP binding sitelac promoterlac operatorM13 revT3 promoterner3xFLAGFRTT7 promoteroriSpectinomycin 9-adenylyltransferaseFRTnerT7 promoterM13 fwdChloramphenicol acetyltransferaseRepAFactor Xa site

pAW068 vector Sequence

LOCUS       V009387                 9406 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009387
VERSION     V009387
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9406)
  AUTHORS   Wilson AC, Perego M, Hoch JA.
  TITLE     New transposon delivery plasmids for insertional mutagenesis in
            Bacillus anthracis
  JOURNAL   J. Microbiol. Methods 71 (3), 332-335 (2007)
   PUBMED   17931726
REFERENCE   2  (bases 1 to 9406)
  AUTHORS   Wilson AC, Perego M, Hoch JA.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-SEP-2007) Department of Molecular and Experimental,
            The Scripps Research Institute, 10550 North Torrey Pines Road, La
            Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 9406)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9406)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Microbiol. Methods"; date: "2007"; volume: "71"; issue: "3"; pages:
            "332-335"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (10-SEP-2007) Department of Molecular and Experimental, The Scripps
            Research Institute, 10550 North Torrey Pines Road, La Jolla, CA
            92037, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             171..1217
                     /codon_start=1
                     /product="C9 transposase"
                     /label="C9 transposase"
                     /protein_id="ABV70027.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     promoter        complement(1280..1298)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1635..1656
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1671..1701
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1709..1725
                     /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     1733..1749
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        1770..1788
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     mobile_element  1801..1827
                     /mobile_element_type="transposon:mariner"
                     /label="ner"
     CDS             1843..1908
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     protein_bind    1912..1959
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(1978..1996)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2483..3071)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             3367..4146
                     /gene="ant1"
                     /label="Spectinomycin 9-adenylyltransferase"
                     /note="Spectinomycin 9-adenylyltransferase from
                     Staphylococcus aureus (strain N315). Accession#: P0A0D1"
     protein_bind    5051..5098
                     /label="FRT"
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     mobile_element  complement(5112..5138)
                     /mobile_element_type="transposon:mariner"
                     /label="ner"
     promoter        complement(5151..5169)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5176..5192)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             5817..6464
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(7080..7778)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label="repA"
                     /protein_id="ABV70030.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(7080..7778)
                     /gene="repA"
                     /label="repA"
     CDS             8390..8401
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"

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