LOCUS       40924_24872        7645 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR-Ech_0379-tuf-aadA, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7645)
  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..7645
                     /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             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
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             5033..5824
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5845..6702
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6876..7464
                     /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 actcgagaaa
     1501 aatgtgacta ttaattttga ctttttttaa ttaggtattg acatattgta atctttaatt
     1561 tagttaaatc aattctaatc taaatttttt tggtatcatt ttttgttttt ttgtgttaat
     1621 taatacaagg atgttaggtt agcgttagtg tttttaggag gagtggccga gcggtcaaag
     1681 gcagcagact gtaaatctgc ccactagcgt gtacgtaggt tcaaatccta cctcctccac
     1741 gggagattat gcgggtatag ctcaatggta gagtcccagc cttccaagct ggctacgtgg
     1801 gttcgattcc cactatccgc tctttatttt tgaaatggtt taatgatgtt aaaggtattt
     1861 gtttatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     1921 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     1981 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     2041 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     2101 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     2161 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     2221 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     2281 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     2341 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     2401 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     2461 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     2521 gtatcagccc gtcatacttg aagctaggca ggcttatctt ggacaagaag atcgcttggc
     2581 ctcgcgcgca gatcagttgg aagaatttgt tcactacgtg aaaggcgaga tcaccaaggt
     2641 agtcggcaaa taactcgagt attatgcttt ataaatgttc tcagtctatt ggcaaagagt
     2701 atactaatga gcaagcaagg tacttacctt gtttttcacc tggttatact cggggtaatg
     2761 ttaagggcaa ttcattcata tgtttaggaa tagctttgtt tatagcacaa gctctattta
     2821 atggatttgg gttattattt atgttgctgt ctgaatttaa gacacaaaat gattggtggt
     2881 ttattggcat atgtgttgtt aatatgctct tagccgttac tgctcttacg ttgatttgta
     2941 ttggaaagaa acaactgagt caaaatcatg ataccttatt aacggatata gatgtcagtg
     3001 gtattaatat tgttgaagaa agtgtagatt tatagttttt tatttgtatg tcatttgtat
     3061 tagatatagc tgataaaaat gcaatttata ttgtaacatg attattaagt ttgatgattt
     3121 attttattat aacaacaagt tatatgtctg tatgtctttc attttatatt tctctgatta
     3181 tataatatca accagttaac aatgtgctct gtatcaattg cttgtcgata tgtttataaa
     3241 atatcagagt gctttttttc tattactgca agcaatcgtt gtgtaggttt ttataagtct
     3301 acttttgatg ttattttact taacttatta tggaaatttc tgttgtctat aaagaatcat
     3361 gtatgtcttt ttacttcttt gtttaattct aaatgatttt attatgtatt gaacattatg
     3421 tgttcttaaa tctatattct taaataaatg tgttatcaat atagcatata ttatcagtag
     3481 atttattgta ttatcaaata gaaagcttaa ttatagaatt tttaaattta aatattattt
     3541 aggtgtgttt gatgcatttt tctaagtaag ttgaatgctt cgctatgtaa gtgagtgtta
     3601 tgtaccctaa taaagtctat accttgttga aatagaaaga ttgatattgc tagtgtagca
     3661 ttgtcacgtg ttggatcatt tgatgtgtat gaagtaagaa agaattttcg tgagtgtcca
     3721 acatatattt tttgatttaa aacttttaat ctatcgatat tgtttaatat ataaaatgtt
     3781 tgattggcgg ttttactaaa tcctatacct ggatcaataa tgattctgtt tttatcaatt
     3841 cctgcatttg ttagtttatc aatgtacttc tttgcccata taataatttc attaattgga
     3901 tcaaggtttt ttggaatagt attgttttta attaatggta tacctagact atgggtaatt
     3961 acaacattta cagaactatt ttgtagcacc tgaagcatgt tcttgtctaa gggcgaattc
     4021 tgcagatatc catcacactg gcggccgctc gagcatgcat ctagagggcc caattcgccc
     4081 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4141 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4201 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4261 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4321 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4381 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4441 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4501 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4561 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4621 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4681 acgcgaattt taacaaaatt cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
     4741 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     4801 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     4861 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     4921 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     4981 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5041 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5101 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5161 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     5221 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5281 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5341 gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5401 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5461 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5521 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     5581 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5641 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5701 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5761 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5821 cttctgaatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5881 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5941 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6001 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6061 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6121 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6181 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6241 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6301 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6361 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6421 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6481 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6541 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6601 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6661 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6721 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6781 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6841 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6961 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7021 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7081 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7141 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7201 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7261 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7321 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7381 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7441 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7501 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7561 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7621 gagtcagtga gcgaggaagc ggaag
//
