LOCUS       Exported                8345 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR-res-Ech_0379-Amtr-mCh-Gent, complete sequence.
ACCESSION   MF068807
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8345)
  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 8345)
  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 8345)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8345
                     /organism="Cloning vector pHR-res-Ech_0379-Amtr-mCh-Gent"
                     /mol_type="other DNA"
                     /label=derived from Ehrlichia chaffeensis str. Arkansas
                     /note="derived from Ehrlichia chaffeensis str. Arkansas"
                     /db_xref="taxon:2056251"
     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             806..1867
                     /codon_start=1
                     /gene="ECH_0379"
                     /product="hypothetical protein"
                     /label=ECH_0379
                     /note="identified by Glimmer2; putative"
                     /protein_id="ATY38628.1"
                     /translation="MAMIIKFGVENLIDSAVEIVCKYFDIYQRNDAVKHVLHKCIKSLR
                     TVEKKLKIYRGKEAKGHVISDSQFAEMEDLIKKVLMKVFEIDISYQKQHLSAEKMEEYK
                     VAVSLLLDQIVRCKSYALKESDILPKVVGNATLKRSAYKCKHVVDFINKNRFLPMNMGR
                     FDKLCFDLDLDPQILKQVIDLCSFALIFEILDLKLVQFLVEMRFFPHQNTNVISITKDN
                     YIELMQHIMLYKCSQSIGKEYTNEQARYLPCFSPGYTRGNVKGNSFICLGIALFIAQAL
                     FNGFGLLFMLLSEFKTQNDWWFIGICVVNMLLAVTALTLICIGKKQLSQNHDTLLTDID
                     VSGINIVEESVDL"
     gene            806..1867
                     /gene="ECH_0379"
                     /label=ECH_0379
                     /note="derived from Ehrlichia chaffeensis str. Arkansas"
     CDS             2096..2806
                     /codon_start=1
                     /gene="mCherry"
                     /product="mCherry fluorescent protein"
                     /label=mCherry
                     /protein_id="ATY38629.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     gene            2096..2806
                     /gene="mCherry"
                     /label=mCherry
     CDS             2817..3350
                     /codon_start=1
                     /product="gentamicin resistance protein"
                     /label=gentamicin resistance protein
                     /note="optimized for Ehrlichia"
                     /protein_id="ATY38630.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(4779..4797)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4804..4820)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4961..5389
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             5733..6527
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin resistance protein"
                     /label=neo
                     /protein_id="ATY38631.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            5733..6527
                     /gene="neo"
                     /label=neo
     CDS             6545..7405
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ATY38632.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            6545..7405
                     /gene="bla"
                     /label=bla
     rep_origin      7576..8164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"