LOCUS       Exported                5824 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNR-46131, complete sequence.
ACCESSION   KP255996
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  AUTHORS   Charpentier E, Anton AI, Barry P, Alfonso B, Fang Y, Novick RP.
  TITLE     Novel cassette-based shuttle vector system for gram-positive 
            bacteria
  JOURNAL   Appl. Environ. Microbiol. 70 (10), 6076-6085 (2004)
  PUBMED    15466553
REFERENCE   2  (bases 1 to 5824)
  AUTHORS   Riojas MA, Bojja K, Farley T, Moreno J, Pederson C, Byrd R, Hamid S,
            Mooney S, Hazbon MH.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-DEC-2014) BEI Bacteriology, BEI Resources/American 
            Type and Culture Collection (ATCC), 10801 University Boulevard, 
            Manassas, VA 20110-2209, USA
REFERENCE   3  (bases 1 to 5824)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Genomics Workbench v. 7.0.2
            Coverage              :: 11,677.82
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5824
                     /organism="Shuttle vector pNR-46131"
                     /mol_type="other DNA"
                     /note="maintained in Staphylococcus aureus in media 
                     supplemented with 10ug/mL erythromycin."
                     /db_xref="taxon:1603882"
     misc_feature    complement(1172..1377)
                     /label=MCS-pUC19 cassette
                     /note="MCS-pUC19 cassette"
     misc_feature    1175..1332
                     /label=Lac operon
                     /note="Lac operon"
     primer_bind     1316..1332
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1333..1377
                     /note="HindIII-SphI-PstI-SalI-XbaI-BamHI-SmaI-KpnI-SacI- 
                     EcoRI; polylinker of M13mp19"
     regulatory      complement(1378..1511)
                     /regulatory_class="promoter"
                     /note="constitutive beta-lactamase promoter; P_blaZ"
     misc_feature    1527..3621
                     /note="E. coli replication and selection cassette; amp 
                     ColE1ori"
     rep_origin      complement(1648..2236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2407..3267)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3268..3372)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    complement(3637..4897)
                     /label=Staphylococcus selection cassette
                     /note="Staphylococcus selection cassette"
     misc_feature    complement(3737..4610)
                     /label=ermC
                     /note="ermC"
     CDS             complement(4899..5204)
                     /codon_start=1
                     /product="truncated polypeptide B"
                     /label=truncated polypeptide B
                     /protein_id="AJF23042.1"
                     /translation="MFSLYKKFKGLFYSVLFWLCILSFFSVLNEMVLNVSLPDIANHFN
                     TTPGITNWVNTAYMLTFSIGTAVYGKLSDYINIKKLLIIGISLSCLGSLIAFIGPT"
     misc_feature    complement(4909..1173)
                     /label=Staphylococcus replication cassette
                     /note="Staphylococcus replication cassette"
     CDS             complement(5366..486)
                     /codon_start=1
                     /product="polypeptide A"
                     /label=polypeptide A
                     /note="RepC protein"
                     /protein_id="AJF23043.1"
                     /translation="MYKNNHANHSNHLENHDLDNFSKTGYSNSRLDAHTVCISDPKLSF
                     DAMTIVGNLNRDNAQALSKFMSVEPQIRLWDILQTKFKAKALQEKVYIEYDKVKADSWD
                     RRNMRIEFNPNKLTRDEMIWLKQNIISYMEDDGFTRLDLAFDFEDDLSDYYAMSDKAVK
                     KTIFYGRNGKPETKYFGVRDSNRFIRIYNKKQERKDNADAEVMSEHLWRVEIELKRDMV
                     DYWNDCFSDLHILQPDWKTIQRTADRAIVFMLLSDEEEWGKLHRNSRTKYKNLIKEISP
                     VDLTDLMKSTLKANEKQLQKQIDFWQHEFKFWK"