LOCUS       Exported                6152 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pSK5640, complete sequence.
ACCESSION   AY182782
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6152)
  AUTHORS   Grkovic S, Brown MH, Hardie KM, Firth N, Skurray RA.
  TITLE     Stable low-copy-number Staphylococcus aureus shuttle vectors
  JOURNAL   Microbiology (Reading, Engl.) 149 (PT 3), 785-794 (2003)
  PUBMED    12634346
REFERENCE   2  (bases 1 to 6152)
  AUTHORS   Grkovic S, Brown MH, Hardie KM, Firth N, Skurray RA.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-NOV-2002) Biological Sciences, University of Sydney, 
            Macleay Building, A12, Sydney, NSW 2006, Australia
REFERENCE   3  (bases 1 to 6152)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6152
                     /organism="Shuttle vector pSK5640"
                     /lab_host="Staphylococcus aureus"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:218964"
     misc_feature    1..57
                     /note="multiple cloning site (MCS); contains uniques sites 
                     for HindIII, SpeI, SalI, XbaI, BamHI, SmaI, and KpnI"
     primer_bind     complement(58..74)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             700..1350
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyl transferase"
                     /function="chloramphenicol resistance"
                     /label=cat
                     /protein_id="AAO24789.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            700..1350
                     /gene="cat"
                     /label=cat
     CDS             complement(1605..2342)
                     /codon_start=1
                     /gene="par"
                     /product="partitioning protein"
                     /label=par
                     /note="Par"
                     /protein_id="AAO24790.1"
                     /translation="MKTIKMVADELNVTKQTVVNNAKNLNISFEKENGVNYIDDNDYLK
                     IVEKITKKERTTQNKENKKSEITYENTEKNRYNNSDGFETLKTKVNELEKQVEIFETRA
                     KNDEKYIENLTKQLDQQNSNVNTLNKLLENQQILALESNKKIQKLEHQLEEERQLSYSF
                     DKSTNDRENFDVQEASYTSDSVNTDQYQKEEKKPEVQPKDISESQQDEKSQQQDDSFNQ
                     NDKDIAIEETQTKKGFWSRLFGG"
     gene            complement(1605..2342)
                     /gene="par"
                     /label=par
     CDS             2767..3687
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /note="Rep"
                     /protein_id="AAO24791.1"
                     /translation="MSKFKKISASEFETLRFYQLPKFLFEDEYFSKMPTDAKVMYALLK
                     DRFELSRLNNWVDSENNIYLLYTNKQLCSILNYAEPKIIKLKKELEKYNLIINERQGLN
                     KPNKIYLLEPTYDKELINSKFQNKEFISSRTNKSSVQELINSKSSDTDFNNTEYIETKN
                     NDTNYTNDTSNMISKNSHSNHTNHQQTEFNNDALKFQALEELPSQIKSYVSNFEIKDIR
                     IIKSILLKGKKSFNNTHDTYYRLEDVEFELVSVLKRFKAMLLQKNETVETMQGYLMQSI
                     KAELEEIHALNMRRQNIPQYNIFNQ"
     gene            2767..3687
                     /gene="rep"
                     /label=rep
     promoter        3783..3887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3888..4748
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin resistance determinant"
                     /label=bla
                     /note="amp"
                     /protein_id="AAO24792.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3888..4748
                     /gene="bla"
                     /label=bla
     rep_origin      4919..5507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5693..5833
                     /label=bom
                     /note="basis of mobility region from pBR322"
     protein_bind    6026..6047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6062..6092
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6100..6116
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     6124..6140
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6136..307
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"