LOCUS       Exported                7645 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR-Ech_0379-tuf-aadA, complete sequence.
ACCESSION   MF068806
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7645)
  AUTHORS   Wang Y, Wei L, Liu H, Cheng C, Ganta RR.
  TITLE     A genetic system for targeted mutations to disrupt and restore genes
            in the obligate bacterium, Ehrlichia chaffeensis
  JOURNAL   Sci Rep 1, 15801 (2017)
  PUBMED    29150636
REFERENCE   2  (bases 1 to 7645)
  AUTHORS   Wang Y, Liu H, Ganta RR.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-MAY-2017) Department of Diagnostic 
            Medicine/Pathobiology, College of Veterinary Medicine, Kansas State 
            University, 1800 Denison Ave, Manhattan, KS 66503, USA
REFERENCE   3  (bases 1 to 7645)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7645
                     /organism="Cloning vector pHR-Ech_0379-tuf-aadA"
                     /mol_type="other DNA"
                     /label=derived from Ehrlichia chaffeensis str. Arkansas
                     /note="derived from Ehrlichia chaffeensis str. Arkansas"
                     /db_xref="taxon:2056250"
     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             1865..2653
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside-3-adenylyltransferase"
                     /label=aadA
                     /protein_id="ATY38625.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            1865..2653
                     /gene="aadA"
                     /label=aadA
     promoter        complement(4079..4097)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4104..4120)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4261..4689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             5033..5827
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin resistance protein"
                     /label=neo
                     /protein_id="ATY38626.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            5033..5827
                     /gene="neo"
                     /label=neo
     CDS             5845..6705
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ATY38627.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5845..6705
                     /gene="bla"
                     /label=bla
     rep_origin      6876..7464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"