LOCUS       Exported                6592 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Staphylococcal shuttle vector pCN56, complete sequence.
ACCESSION   KR781471
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6592)
  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  JOURNAL   Unpublished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 6592)
  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 6592)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Genomics Workbench v. 7.0.2
            Coverage              :: 38389
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6592
                     /organism="Staphylococcal shuttle vector pCN56"
                     /mol_type="other DNA"
                     /clone="NRS621"
                     /note="maintained in Escherichia coli grown in media 
                     supplemented with 100 ug/ml ampicillin"
                     /db_xref="taxon:1740137"
                     /culture_collection="BEI:NARSA NR-46156"
     misc_feature    1..2087
                     /note="T181cop-623 repC; Staphylococcus replication 
                     cassette; high-copy-number variant; derived from J04764"
     misc_feature    158..391
                     /label=RS-B site
                     /note="RS-B site"
     rep_origin      654..821
                     /note="ori; derived from Staphylococcus aureus"
     CDS             686..1630
                     /codon_start=1
                     /product="RepC protein"
                     /label=RepC protein
                     /note="polypeptide A"
                     /protein_id="AMR43698.1"
                     /translation="MYKNNHANHSNHLENHDLDNFSKTGYSNSRLDAHTVCISDPKLSF
                     DAMTIVGNLNRDNAQALSKFMSVEPQIRLWDILQTKFKAKALQEKVYIEYDKVKADSWD
                     RRNMRIEFNPNKLTRDEMIWLKQNIISYMEDDGFTRLDLAFDFEDDLSDYYAMSDKAVK
                     KTIFYGRNGKPETKYFGVRDSNRFIRIYNKKQERKDNADAEVMSEHLWRVEIELKRDMV
                     DYWNDCFSDLHILQPDWKTIQRTADRAIVFMLLSDEEEWGKLHRNSRTKYKNLIKEISP
                     VDLTDLMKSTLKANEKQLQKQIDFWQHEFKFWK"
     variation       1130..1132
                     /label=E149D
                     /note="E149D"
     misc_feature    1792..2087
                     /label=truncated polypeptide B
                     /note="truncated polypeptide B"
     misc_feature    2099..3359
                     /note="pE194 ermC; erythromycin resistance; Staphylococcus 
                     selection cassette; derived from J01755"
     misc_feature    2406..2465
                     /label=leader peptide
                     /note="leader peptide"
     CDS             2525..3259
                     /codon_start=1
                     /product="adenine methylase"
                     /label=adenine methylase
                     /note="ermC"
                     /protein_id="AMR43699.1"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
                     NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     misc_feature    3376..5469
                     /note="amp ColE1 ori; Escherichia coli replication and 
                     selection cassette; derived from L09137"
     promoter        3624..3728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3729..4589
                     /codon_start=1
                     /product="beta lactamase"
                     /label=beta lactamase
                     /note="involved in ampicillin resistance"
                     /protein_id="AMR43697.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4760..5348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(4964..5244)
                     /direction=LEFT
                     /note="ori; derived from Escherichia coli"
     misc_feature    5485..5529
                     /note="polylinker; derived from M13mp19"
     misc_feature    5550..6279
                     /note="gfpmut2; FACS-optimized GFP mutant; derived from 
                     U73901"
     CDS             5563..6279
                     /codon_start=1
                     /product="GFPmut2"
                     /label=GFPmut2
                     /note="green fluorescent protein mutant; FACS-optimized GFP
                     mutant"
                     /protein_id="AMR43700.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     variation       5755..5757
                     /label=S65A
                     /note="S65A"
     variation       5764..5766
                     /label=V68L
                     /note="V68L"
     variation       5776..5778
                     /label=S72A
                     /note="S72A"
     misc_feature    6288..6588
                     /label=blaZTT
                     /note="blaZTT"
     misc_feature    6451..6588
                     /label=TT
                     /note="TT"