LOCUS       Exported                5544 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNR-46121, complete sequence.
ACCESSION   KM015349
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5544)
  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 5544)
  AUTHORS   Riojas MA, Bojja K, Hazbon MH.
  TITLE     pNR-46121, A Shuttle Vector for Gram-Positive Bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5544)
  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 5544)
  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..5544
                     /organism="Shuttle vector pNR-46121"
                     /mol_type="other DNA"
                     /db_xref="taxon:1539884"
     misc_feature    1..1417
                     /note="pT181 cop-wt repC; derived from J01764.1 
                     Staphylococcus aureus plasmid pT181"
     CDS             157..960
                     /codon_start=1
                     /gene="repC"
                     /product="replication initiation protein"
                     /label=repC
                     /protein_id="AIL56502.1"
                     /translation="MTIVGNLNRDNAQALSKFMSVEPQIRLWDILQTKFKAKALQEKVY
                     IEYDKVKADSWDRRNMRIEFNPNKLTRDEMIWLKQNIISYMEDDGFTRLDLAFDFEDDL
                     SDYYAMSDKAVKKTIFYGRNGKPETKYFGVRDSNRFIRIYNKKQERKDNADAEVMSEHL
                     WRVEIELKRDMVDYWNDCFSDLHILQPDWKTIQRTADRAIVFMLLSDEEEWGKLHRNSR
                     TKYKNLIKEISPVDLTDLMKSTLKANEKQLQKQIDFWQHEFKFWK"
     gene            157..960
                     /gene="repC"
                     /label=repC
     misc_feature    1122..1417
                     /gene="tetK"
                     /label=truncated tetracycline efflux protein
                     /note="truncated tetracycline efflux protein"
     gene            1122..1417
                     /gene="tetK"
                     /label=tetK
     misc_feature    1429..2545
                     /note="aphA-3; derived from V01547.1 Streptococcus faecalis
                     kanamycin resistance gene"
     CDS             1611..2405
                     /codon_start=1
                     /gene="kanR"
                     /product="kanamycin resistance protein"
                     /label=kanR
                     /protein_id="AIL56503.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            1611..2405
                     /gene="kanR"
                     /label=kanR
     misc_feature    2559..4653
                     /note="amp ColE1ori; derived from L09137.2 Cloning vector 
                     pUC19c"
     promoter        2808..2912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2913..3773
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin resistance protein"
                     /label=bla
                     /protein_id="AIL56504.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2913..3773
                     /gene="bla"
                     /label=bla
     rep_origin      3944..4532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4044..4534
                     /label=ColE
                     /note="ColE"
     misc_feature    4663..4874
                     /note="MCS-pUC19; derived from L09137.2 Cloning vector 
                     pUC19c"
     primer_bind     complement(4714..4730)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4873..5544
                     /note="pT181 cop-wt repC; derived from J01764.1 
                     Staphylococcus aureus plasmid pT181"