LOCUS       40924_24877        8345 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR-res-Ech_0379-Amtr-mCh-Gent, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8345)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep 1, 
            15801 (2017)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-MAY-2017) Department of Diagnostic Medicine/Pathobiology, 
            College of Veterinary Medicine, Kansas State University, 1800 
            Denison Ave, Manhattan, KS 66503, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /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
                     /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..2803
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             2817..3347
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /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
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             5733..6524
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6545..7402
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7576..8164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc ccttacctgc
      301 tgtactgagt atgttcttgg tatactgttg atatgaaatg tattattgaa agtagtaaat
      361 tcttgattag gatgagaagt actatgattg taaattggtg ttatgtttat ctctttttat
      421 gaggaaagta tagctaatgt tataaatgaa tagaatatat tgaaaagcta tgcttagtat
      481 gtgaatgttt tgtacggtag gtatgaattt aatataatat gttttttagc atcctttgtg
      541 ttaaaagtaa tattgtcatt ttataacttc ctgattatta ctgtattttt tcagaatatg
      601 actagtgtaa tattttaagt tctcatatta ttaatacaat attaaatact aaattaaata
      661 tattaaaaaa ttattacaaa taattaagaa ttagtaaact ttatatgttt atgacattta
      721 tagaaaatgt tatttttaca actcttagtt gtatagtata aaaattatta tactatacta
      781 atttaataat aaataggtaa gtaagatggc tatgataata aagtttgggg ttgaaaatct
      841 gatcgatagt gctgtggaaa tagtttgtaa atattttgat atatatcagc gaaatgatgc
      901 agtaaaacat gtattacata aatgtattaa aagtttaaga actgtagaaa agaaattaaa
      961 gatatataga gggaaagaag ctaaaggaca tgtaattagt gatagccaat ttgctgaaat
     1021 ggaggatcta ataaaaaaag ttctcatgaa ggtgtttgag attgatattt cttatcaaaa
     1081 acaacacttg tctgcagaaa aaatggagga gtataaagta gctgttagtc ttttattgga
     1141 tcaaatagta aggtgtaagt cttatgctct taaagaaagt gatattcttc ctaaggttgt
     1201 agggaatgca accttaaaac gtagtgcata taagtgtaag catgtagtgg attttattaa
     1261 taagaataga tttcttccta tgaatatggg gagatttgat aagctttgtt ttgatctaga
     1321 tttagatcct cagattttga aacaagttat agatttatgc agttttgctc ttatatttga
     1381 aattttggat cttaagttag tacaatttct tgttgaaatg agattctttc cacatcaaaa
     1441 cactaatgta attagtataa ctaaagataa ttacatagaa ttaatgcagc atattatgct
     1501 ttataaatgt tctcagtcta ttggcaaaga gtatactaat gagcaagcaa ggtacttacc
     1561 ttgtttttca cctggttata ctcggggtaa tgttaagggc aattcattca tatgtttagg
     1621 aatagctttg tttatagcac aagctctatt taatggattt gggttattat ttatgttgct
     1681 gtctgaattt aagacacaaa atgattggtg gtttattggc atatgtgttg ttaatatgct
     1741 cttagccgtt actgctctta cgttgatttg tattggaaag aaacaactga gtcaaaatca
     1801 tgatacctta ttaacggata tagatgtcag tggtattaat attgttgaag aaagtgtaga
     1861 tttatagctc ctagaacgat cgccgcatgc tagcataaaa cgcgcgcact aacaagtgcc
     1921 cagcgcgctg cctcaaaaaa tctccggaag tttcccggat atttattgat gattttattt
     1981 ttaatctagc ataccattta cagttttatt attacaacca tacattaaat aactgttgca
     2041 ttaacattat gtatgattta tcctaagtta tctgagtaac ataagggata ttataatggt
     2101 cagcaaggga gaggaagata atatggcgat catcaaagag tttatgagat ttaaggtgca
     2161 catggaagga agcgttaatg gtcatgagtt tgaaatcgaa ggcgaaggcg aaggaagacc
     2221 gtatgaaggc acacagacgg ctaaacttaa ggtcacaaaa ggcggaccgc ttccattcgc
     2281 gtgggatatt ctttcaccgc aatttatgta tggttctaaa gcctatgtga aacatcctgc
     2341 ggatattcct gactacctta aactgtcttt cccggaagga tttaaatggg aacgcgtcat
     2401 gaacttcgaa gatggcggcg ttgttacggt gacgcaggat tcatcactgc aagatggaga
     2461 atttatttat aaagttaaac tgcgcggcac aaactttccg tcagacggac ctgtcatgca
     2521 gaagaaaacg atgggctggg aagccagcag cgagagaatg tacccggagg acggagcact
     2581 taaaggcgaa atcaagcaac gcctgaagct gaaagatgga ggccattatg atgccgaggt
     2641 caagacgaca tacaaagcta agaaaccggt acaattacct ggagcataca acgtcaatat
     2701 caagctggat attacgtcac ataatgaaga ctatacgatt gtagagcaat atgaaagagc
     2761 agagggaaga cactctacag gtggaatgga cgaattatac aaatagggat taaattatgt
     2821 taagatcatc aaatgatgtt acacaacaag gaagtagacc aaaaactaaa ttaggtggta
     2881 gtagtatggg aattattaga acttgtagat taggtcctga tcaagtaaaa agtatgagag
     2941 cagctttaga tttatttggt agagaatttg gagatgttgc aacatattct caacatcaac
     3001 cagatagtga ttatttgggt aatttattaa gatcaaaaac ttttatagct ttagcagctt
     3061 ttgatcaaga agcagtagtt ggagctttag cagcttatgt attacctaga tttgaacaac
     3121 caagatcaga aatttatata tatgatttag cagtttctgg tgaacataga agacaaggaa
     3181 tagcaacagc tttaataaat ttattaaaac atgaagcaaa tgctttaggt gcatatgtaa
     3241 tttatgttca agctgattat ggagatgatc ctgcagtagc tttatatact aaattaggta
     3301 ttagagaaga agttatgcat tttgatattg atccaagtac agcaacataa actagtagtt
     3361 attatgcttt ataaatgttc tcagtctatt ggcaaagagt atactaatga gcaagcaagg
     3421 tacttacctt gtttttcacc tggttatact cggggtaatg ttaagggcaa ttcattcata
     3481 tgtttaggaa tagctttgtt tatagcacaa gctctattta atggatttgg gttattattt
     3541 atgttgctgt ctgaatttaa gacacaaaat gattggtggt ttattggcat atgtgttgtt
     3601 aatatgctct tagccgttac tgctcttacg ttgatttgta ttggaaagaa acaactgagt
     3661 caaaatcatg ataccttatt aacggatata gatgtcagtg gtattaatat tgttgaagaa
     3721 agtgtagatt tatagttttt tatttgtatg tcatttgtat tagatatagc tgataaaaat
     3781 gcaatttata ttgtaacatg attattaagt ttgatgattt attttattat aacaacaagt
     3841 tatatgtctg tatgtctttc attttatatt tctctgatta tataatatca accagttaac
     3901 aatgtgctct gtatcaattg cttgtcgata tgtttataaa atatcagagt gctttttttc
     3961 tattactgca agcaatcgtt gtgtaggttt ttataagtct acttttgatg ttattttact
     4021 taacttatta tggaaatttc tgttgtctat aaagaatcat gtatgtcttt ttacttcttt
     4081 gtttaattct aaatgatttt attatgtatt gaacattatg tgttcttaaa tctatattct
     4141 taaataaatg tgttatcaat atagcatata ttatcagtag atttattgta ttatcaaata
     4201 gaaagcttaa ttatagaatt tttaaattta aatattattt aggtgtgttt gatgcatttt
     4261 tctaagtaag ttgaatgctt cgctatgtaa gtgagtgtta tgtaccctaa taaagtctat
     4321 accttgttga aatagaaaga ttgatattgc tagtgtagca ttgtcacgtg ttggatcatt
     4381 tgatgtgtat gaagtaagaa agaattttcg tgagtgtcca acatatattt tttgatttaa
     4441 aacttttaat ctatcgatat tgtttaatat ataaaatgtt tgattggcgg ttttactaaa
     4501 tcctatacct ggatcaataa tgattctgtt tttatcaatt cctgcatttg ttagtttatc
     4561 aatgtacttc tttgcccata taataatttc attaattgga tcaaggtttt ttggaatagt
     4621 attgttttta attaatggta tacctagact atgggtaatt acaacattta cagaactatt
     4681 ttgtagcacc tgaagcatgt tcttgtctaa gggcgaattc tgcagatatc catcacactg
     4741 gcggccgctc gagcatgcat ctagagggcc caattcgccc tatagtgagt cgtattacaa
     4801 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     4861 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     4921 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
     4981 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5041 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5101 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5161 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5221 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5281 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5341 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaatt
     5401 cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt
     5461 gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
     5521 agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat
     5581 ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
     5641 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat
     5701 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5761 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5821 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5881 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5941 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     6001 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatcccac cttgctcctg
     6061 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6121 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6181 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6241 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6301 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6361 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6421 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6481 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt gaaaaaggaa
     6541 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6601 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6661 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6721 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6781 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6841 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6901 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6961 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7021 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7081 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7141 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7201 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7261 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7321 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7381 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7441 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7501 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7561 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7621 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7681 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7741 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7801 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7861 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7921 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7981 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8041 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8101 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8161 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8221 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     8281 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     8341 ggaag
//
