LOCUS       Exported                6409 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pBSV2, complete sequence.
ACCESSION   AY187276
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6409)
  AUTHORS   Stewart PE, Thalken R, Bono JL, Rosa P.
  TITLE     Isolation of a circular plasmid region sufficient for autonomous 
            replication and transformation of infectious Borrelia burgdorferi
  JOURNAL   Mol. Microbiol. 39 (3), 714-721 (2001)
  PUBMED    11169111
REFERENCE   2  (bases 1 to 6409)
  AUTHORS   Stewart PE.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-NOV-2002) LHBP, RML, 903 South 4th Street, Hamilton, 
            MT 59840, USA
REFERENCE   3  (bases 1 to 6409)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6409
                     /organism="Shuttle vector pBSV2"
                     /lab_host="Borrelia burgdorferi"
                     /mol_type="genomic DNA"
                     /note="Borrelia burgdorferi/Escherichia coli shuttle vector
                     conferring kanamycin resistance"
                     /db_xref="taxon:219746"
     rep_origin      1..714
                     /label=ColE1
                     /note="ColE1"
     rep_origin      71..659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     repeat_region   790..973
                     /label=IR2
                     /note="IR2"
     CDS             complement(1178..1738)
                     /codon_start=1
                     /product="Orf3"
                     /label=Orf3
                     /protein_id="AAO32125.1"
                     /translation="MEKKRVVKVLTKKIDTYVEQNLMINESKISYYKTLKEKLNDNFKK
                     EIFHRVENIKILKEIKDNQYYKFDGYKTFLDFIKDFDVAKTQAYKYLRLATALQEGLIK
                     EDYLIENGIKNSYNFIKDKESPALKKSRQNPIKPLRFQLKTQESYDFYKKKSKFTSFMM
                     HEIFENQKDFLKKLLKKYEELKI"
     CDS             complement(1790..2341)
                     /codon_start=1
                     /product="Orf2"
                     /label=Orf2
                     /protein_id="AAO32124.1"
                     /translation="MKIGLKAARINEQKIKFPKESIFLLKEKKGDKNIYHTKILKALHR
                     VVSNKQKRCIIFFEKLLDKSEIQKLYLYPMKDGDKFLGIYYGYRKPINNVFVKYEINGV
                     ENVYTFSKIYYIEFRFKTGSVCCYLKNMRRLLRKEKIDTPYNKAIVDKFIDLEKHVYEF
                     YNKKYSSEGLILKWILKNLK"
     CDS             complement(2351..3478)
                     /codon_start=1
                     /product="Orf1"
                     /label=Orf1
                     /protein_id="AAO32123.1"
                     /translation="MENLSNINKTSKTTTCYNKLQYKLVVLISTLAYINKTYKKYTQKN
                     ILYYFNENLKRNSQTPVKLKTLQNYLYKLEKEFKVTTNYHKHLGVNLGTEIYYQLNFSK
                     KECYQKIYKYFQEKKDLRFQNRVSRGLKDRFTKNGSVDLKECLNNKNNIKEERKINEIE
                     KYQVRNYFNKCNFLCKKILSIFLTILFNLNIDKDNIIKILKIIKIIEIKLLKNKNIHFT
                     KSCMKEKQEKLKEILCNTQKEFEKNEYNSKQLETSFQKIYENYKFKPHFIIESHKYSDL
                     NNIKRKLEKSIERKKENSQQNYQDLKTNIFNILIEQLKKEVNIELLKPIIKEYLNNQKK
                     IEYNKVFCTYYCELLELMKNQKSLLSLKELNRKAI"
     repeat_region   3709..3892
                     /label=IR1
                     /note="IR1"
     misc_feature    4031..4491
                     /label=MCS-LacZ derived from pUC19
                     /note="MCS-LacZ derived from pUC19"
     protein_bind    4142..4163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4178..4208
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4216..4232
                     /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     4240..4256
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4252..4542
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRGRYPSFKEDFLLRLNLRA"
     misc_feature    4266..4322
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4326..4342)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4492..4898
                     /regulatory_class="promoter"
                     /note="FlgB promoter"
     CDS             4899..5708
                     /codon_start=1
                     /gene="Kan"
                     /product="kanamycin"
                     /label=Kan
                     /protein_id="AAO32126.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            4899..5708
                     /gene="Kan"
                     /label=Kan
     regulatory      5962..5971
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5968..6342
                     /codon_start=1
                     /product="zeocin"
                     /label=zeocin
                     /note="Zeo"
                     /protein_id="AAO32127.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"