LOCUS       Exported                7179 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Staphylococcal shuttle vector pCN54, complete sequence.
ACCESSION   KR781470
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7179)
  AUTHORS   Riojas MA, Bojja K, Byrd R, Hamid S, Mooney S, Monaco A, Alalade R, 
            King S, Hazbon MH.
  TITLE     Sequencing of a set of cassette-based shuttle vectors for 
            Gram-positive bacteria deposited at BEI resources from the NARSA 
            collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7179)
  AUTHORS   Riojas MA, Bojja K, Byrd R, Hamid S, Mooney S, Monaco A, Alalade R, 
            King S, Hazbon MH.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-MAY-2015) BEI Bacteriology, BEI Resources/American 
            Type and Culture Collection (ATCC), 10801 University Boulevard, 
            Manassas, VA 20110-2209, USA
REFERENCE   3  (bases 1 to 7179)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Genomics Workbench v. 7.0.2
            Coverage              :: 36580
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7179
                     /organism="Staphylococcal shuttle vector pCN54"
                     /mol_type="other DNA"
                     /clone="NRS619"
                     /note="maintained in Escherichia coli grown in media 
                     supplemented with 100 ug/ml ampicillin"
                     /db_xref="taxon:1740135"
                     /culture_collection="BEI:NARSA NR-46154"
     misc_feature    1..2089
                     /note="pT181copwt repC; Staphylococcus replication 
                     cassette; derived from J04764"
     misc_feature    159..392
                     /label=RS-B site
                     /note="RS-B site"
     CDS             688..1632
                     /codon_start=1
                     /product="RepC protein"
                     /label=RepC protein
                     /note="polypeptide A"
                     /protein_id="AMR43693.1"
                     /translation="MYKNNHANHSNHLENHDLDNFSKTGYSNSRLDAHTVCISDPKLSF
                     DAMTIVGNLNRDNAQALSKFMSVEPQIRLWDILQTKFKAKALQEKVYIEYDKVKADSWD
                     RRNMRIEFNPNKLTRDEMIWLKQNIISYMEDDGFTRLDLAFDFEDDLSDYYAMSDKAVK
                     KTIFYGRNGKPETKYFGVRDSNRFIRIYNKKQERKDNADAEVMSEHLWRVEIELKRDMV
                     DYWNDCFSDLHILQPDWKTIQRTADRAIVFMLLSDEEEWGKLHRNSRTKYKNLIKEISP
                     VDLTDLMKSTLKANEKQLQKQIDFWQHEFKFWK"
     variation       1132..1134
                     /label=E149D
                     /note="E149D"
     misc_feature    1794..2089
                     /label=truncated polypeptide B
                     /note="truncated polypeptide B"
     misc_feature    2101..3361
                     /note="pE194 ermC; erythromycin resistance; Staphylococcus 
                     selection cassette; derived from J01755"
     misc_feature    2408..2467
                     /label=leader peptide
                     /note="leader peptide"
     CDS             2527..3261
                     /codon_start=1
                     /gene="ermC"
                     /product="adenine methylase"
                     /label=ermC
                     /protein_id="AMR43694.1"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
                     NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     gene            2527..3261
                     /gene="ermC"
                     /label=ermC
     misc_feature    3378..5471
                     /note="amp ColE1 ori; Escherichia coli replication and 
                     selection cassette; derived from L09137"
     promoter        3626..3730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3731..4591
                     /codon_start=1
                     /product="beta lactamase"
                     /label=beta lactamase
                     /note="involved in ampicillin resistance"
                     /protein_id="AMR43695.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4762..5350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(4966..5246)
                     /direction=LEFT
                     /note="ori; derived from Escherichia coli"
     regulatory      5487..6072
                     /regulatory_class="promoter"
                     /note="Pcad; cadmium-inducible promoter; derived from 
                     J04551"
     regulatory      5996..6001
                     /regulatory_class="minus_35_signal"
                     /note="cadA -35"
     regulatory      6019..6024
                     /regulatory_class="minus_10_signal"
                     /note="cadA -10"
     misc_feature    6073..6117
                     /note="polylinker; derived from M13mp19"
     misc_feature    6138..6867
                     /note="gfpmut2; FACS-optimized GFP mutant; derived from 
                     U73901"
     CDS             6151..6867
                     /codon_start=1
                     /product="GFPmut2"
                     /label=GFPmut2
                     /note="mutant green fluorescent protein"
                     /protein_id="AMR43696.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     variation       6343..6345
                     /label=S65A
                     /note="S65A"
     variation       6352..6354
                     /label=V68L
                     /note="V68L"
     variation       6364..6366
                     /label=S72A
                     /note="S72A"
     misc_feature    6876..7174
                     /label=blaZ-TT
                     /note="blaZ-TT"
     misc_feature    7038..7174
                     /label=TT
                     /note="TT"