LOCUS       Exported                7788 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNR-46124, complete sequence.
ACCESSION   KM015350
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7788)
  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 7788)
  AUTHORS   Riojas MA, Bojja K, Hazbon MH.
  TITLE     pNR-46124, A Shuttle Vector for Gram-Positive Bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7788)
  AUTHORS   Riojas MA, Bojja K, Hazbon MH.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2014) Bacteriology, BEI Resources/American Type 
            Culture Collection (ATCC), 10801 University Blvd., Manassas, VA 
            20110, USA
REFERENCE   4  (bases 1 to 7788)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Genomics Workbench v. 7.0.2
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7788
                     /organism="Shuttle vector pNR-46124"
                     /mol_type="other DNA"
                     /db_xref="taxon:1539885"
     misc_feature    7..2589
                     /note="tetM; derived from M21136.1 S.aureus tetracycline 
                     resistance (tetM) gene"
     CDS             295..2229
                     /codon_start=1
                     /gene="tetM"
                     /product="tetracycline resistance protein"
                     /label=tetM
                     /protein_id="AIL56505.1"
                     /translation="MEENHMKIINIGVLAHVDAGKTTLTESLLYNSGAITELGSVDKGT
                     TRTDNTLLERQRGITIQTGITSFQWENTKVNIIDTPGHMDFLAEVYRSLSVLDGAILLI
                     SAKDGVQAQTRILFHALRKMGIPTIFFINKIDQNGIDLSTVYQDIKEKLSAEIVIKQKV
                     ELYPNMCVTNFTESEQWDTVIEGNDDLLEKYMSGKSLEALELEQEESIRFQNCSLFPLY
                     HGSAKSNIGIDNLIEVITNKFYSSTHRGPSELCGNVFKIEYTKKRQRLAYIRLYSGVLH
                     LRDSVRVSEKEKIKVTEMYTSINGELCKIDRAYSGEIVILQNEFLKLNSVLGDTKLLPQ
                     RKKIENPHPLLQTTVEPSKPEQREMLLDALLEISDSDPLLRYYVDSTTHEIILSFLGKV
                     QMEVISALLQEKYHVEIELKEPTVIYMERPLKNAEYTIHIEVPPNPFWASIGLSVSPLP
                     LGSGMQYESSVSLGYLNQSFQNAVMEGIRYGCEQGLYGWNVTDCKICFKYGLYYSPVST
                     PADFRMLTPIVLEQAFRKAGTELLEPYLSFKVYAPQEYLSRAYNDAPKYCANIVNTQLK
                     NNEVIIIGEIPARCIQDYRNDLTFFTNGLSVCLAELKGYQVTTGEPVCQTRRLNSRIDK
                     VRYMFNKIT"
     gene            295..2229
                     /gene="tetM"
                     /label=tetM
     misc_feature    2604..4698
                     /note="amp ColE1ori; derived from L09137.2 Cloning vector 
                     pUC19c"
     promoter        2853..2957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2958..3818
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin resistance protein"
                     /label=bla
                     /protein_id="AIL56506.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2958..3818
                     /gene="bla"
                     /label=bla
     rep_origin      3989..4577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4089..4579
                     /label=ColE
                     /note="ColE"
     misc_feature    4708..4919
                     /note="MCS-pUC19; derived from L09137.2 Cloning vector 
                     pUC19c"
     primer_bind     complement(4759..4775)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(4918..6981,7750..7783)
                     /note="pT181 cop-wt repC; derived from J01764.1 
                     Staphylococcus aureus plasmid pT181"
     CDS             5605..6549
                     /codon_start=1
                     /gene="repC"
                     /product="replication initiation protein"
                     /label=repC
                     /protein_id="AIL56507.1"
                     /translation="MYKNNHANHSNHLENHDLDNFSKTGYSNSRLDAHTVCISDPKLSF
                     DAMTIVGNLNRDNAQALSKFMSVEPQIRLWDILQTKFKAKALQEKVYIEYDKVKADSWD
                     RRNMRIEFNPNKLTRDEMIWLKQNIISYMEDDGFTRLDLAFDFEDDLSDYYAMSDKAVK
                     KTIFYGRNGKPETKYFGVRDSNRFIRIYNKKQERKDNADAEVMSEHLWRVEIELKRDMV
                     DYWNDCFSDLHILQPDWKTIQRTADRAIVFMLLSDEEEWGKLHRNSRTKYKNLIKEISP
                     VDLTDLMKSTLKANEKQLQKQIDFWQHEFKFWK"
     gene            5605..6549
                     /gene="repC"
                     /label=repC
     misc_feature    join(6711..6981,7750..7783)
                     /gene="tetK"
                     /note="truncated tetracycline efflux protein; interrupted 
                     by E. coli IS1 transposon InsB1"
     gene            6711..7783
                     /gene="tetK"
                     /label=tetK
     mobile_element  complement(6982..7749)
                     /mobile_element_type="insertion sequence:IS1"
                     /note="derived from KJ494659 transposon insertion sequence 
                     (IS1)"
     CDS             complement(6997..7500)
                     /codon_start=1
                     /gene="insB"
                     /product="transposase B"
                     /label=insB
                     /protein_id="AIL56508.1"
                     /translation="MPGNSPHYGRWPQHDFTSLKKLRPQSVTSRIQPGSDVIVCAEMDE
                     QWGYVGAKSRQRWLFYAYDSLRKTVVAHVFGERTMATLGRLMSLLSPFDVVIWMTDGWP
                     LYESRLKGKLHVISKRYTQRIERHNLNLRQHLARLGRKSLSFSKSVELHDKVIGHYLNI
                     KHYQ"
     gene            complement(6997..7500)
                     /gene="insB"
                     /label=insB
     CDS             complement(7419..7694)
                     /codon_start=1
                     /gene="insA"
                     /product="transposase"
                     /label=insA
                     /protein_id="AIL56509.1"
                     /translation="MASVSISCPSCSATDGVVRNGKSTAGHQRYLCSHCRKTWQLQFTY
                     TASQPGTHQKIIDMAMNGVGCRATARIMGVGLNTILRHLKNSGRSR"
     gene            complement(7419..7694)
                     /gene="insA"
                     /label=insA