LOCUS       Exported                4919 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCR2.1-kanR, complete sequence.
ACCESSION   EF488744
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4919)
  AUTHORS   Battisti JM, Raffel SJ, Schwan TG.
  TITLE     A System for Site-Specific Genetic Manipulation of the Relapsing 
            Fever Spirochete Borrelia hermsii
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4919)
  AUTHORS   Battisti JM, Raffel SJ, Schwan TG.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2007) Laboratory of Zoonotic Pathogens, Rocky 
            Mountain Laboratories, National Institute of Allergy and Infectious 
            Diseases, National Institutes of Health, 903 S. 4th Street, 
            Hamilton, MT 59840, USA
REFERENCE   3  (bases 1 to 4919)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4919
                     /organism="Cloning vector pCR2.1-kanR"
                     /mol_type="other DNA"
                     /db_xref="taxon:456496"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(369..1178)
                     /codon_start=1
                     /gene="kanR"
                     /product="kanamycin resistance protein"
                     /label=kanR
                     /protein_id="ABS71236.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(369..1178)
                     /gene="kanR"
                     /label=kanR
     regulatory      complement(1179..1299)
                     /regulatory_class="promoter"
                     /note="flgB promoter derived from Borrelia hermsii"
     promoter        complement(1376..1394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1401..1417)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1559..1939
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             2307..3101
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      4150..4738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"