LOCUS       Exported                7468 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRLBCR8, complete sequence.
ACCESSION   JQ768524
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7468)
  AUTHORS   Redder P, Linder P.
  TITLE     New Range of Vectors with a Stringent 5-Fluoroorotic Acid-Based 
            Counterselection System for Generating Mutants by Allelic 
            Replacement in Staphylococcus aureus
  JOURNAL   Appl. Environ. Microbiol. 78 (11), 3846-3854 (2012)
  PUBMED    22447609
REFERENCE   2  (bases 1 to 7468)
  AUTHORS   Redder P, Linder P.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-MAR-2012) Department of Microbiology and Molecular 
            Medicine, University of Geneva, Faculty of Medicine, Rue 
            Michel-Servet 1, Geneva 1203, Switzerland
REFERENCE   3  (bases 1 to 7468)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: BioEdit, CAP v. 7.0.9.0
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7468
                     /organism="Cloning vector pRLBCR8"
                     /mol_type="other DNA"
                     /db_xref="taxon:1193158"
     misc_feature    14..3371
                     /label=derived from pC194 Staphylococcus aureus plasmid
                     /note="derived from pC194 Staphylococcus aureus plasmid"
     rep_origin      complement(50..1669)
                     /direction=LEFT
                     /note="thermosensitive origin of replication; for 
                     replication in Staphylococcus aureus.; pT181ts"
     misc_feature    1670..2325
                     /note="fragment of the pyrFE genes from Staphylococcus 
                     aureus; replacing the pC194 origin of replication"
     misc_feature    2326..3328
                     /note="inserted between BsrGI and BstBI; CAT194 cassette"
     misc_feature    3372..3505
                     /note="multiple cloning site; MCS"
     misc_feature    3399..3506
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3416..3432
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3466..3482)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3515..3533)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3542..7468
                     /note="dervied from pGB2; pSC101 based E. coli vector with 
                     spectinomycin resistance; no significant similarity to 
                     pBR322-based vectors; point mutation to increase copy 
                     number; pGB2 backbone increased copy number"
     rep_origin      3982..4204
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4252..5202
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     misc_feature    4528
                     /note="G to A mutation in the pGB2 backbone for increased 
                     copy number in E. coli"
     CDS             6454..7245
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"