pLSP2 vector (V004932)

LOCUS       40924_28761        3792 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLSP2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3792)
  AUTHORS   Lehman SS, Mladinich KM, Boonyakanog A, Mima T, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     Versatile nourseothricin and streptomycin/spectinomycin resistance 
            gene cassettes and their use in chromosome integration vectors
  JOURNAL   J. Microbiol. Methods 129, 8-13 (2016)
  PUBMED    27457407
REFERENCE   2  (bases 1 to 3792)
  AUTHORS   Lehman SS, Mladinich K, Boonyakanog A, Mima T, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-MAR-2016) Molecular Genetics and Microbiology, 
            University of Florida, 2055 Mowry Road, Gainesville, FL 32610, USA
REFERENCE   3  (bases 1 to 3792)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3792)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods 129, 8-13 (2016)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-MAR-2016) Molecular Genetics and Microbiology, University of 
            Florida, 2055 Mowry Road, Gainesville, FL 32610, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(946..1803)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1804..1908)
                     /label=AmpR promoter
     primer_bind     2380..2396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2400..2444
                     /note="MCS; multiple cloning site"
     misc_feature    2450..2482
                     /note="loxP; Cre recombinase site"
     regulatory      2550..2603
                     /label=chloramphenicol transacetylase gene promoter
                     /note="chloramphenicol transacetylase gene promoter"
                     /regulatory_class="promoter"
     regulatory      2567..2572
                     /regulatory_class="minus_35_signal"
     regulatory      2590..2595
                     /regulatory_class="minus_10_signal"
     regulatory      2637..2641
                     /regulatory_class="ribosome_binding_site"
     CDS             2649..3401
                     /codon_start=1
                     /gene="aad9"
                     /product="spectinomycin adenyltransferase"
                     /label=aad9
                     /protein_id="ANY60775.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            2649..3401
                     /gene="aad9"
                     /label=aad9
                     /note="codon optimized; derived from Enterococcus faecalis"
     regulatory      3402..3443
                     /regulatory_class="terminator"
     protein_bind    complement(3479..3512)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3517..3561
                     /note="MCS; multiple cloning site"
     primer_bind     complement(3574..3590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3598..3614)
                     /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(3622..3652)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3667..3688)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."