LOCUS       40924_24867        7173 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR-Ech_0230-tuf-aadA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7173)
  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 7173)
  AUTHORS   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 7173)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7173)
  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..7173
                     /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"
     misc_feature    295..1333
                     /label=Ehrlichia chaffeensis str. Arkansas genomic region
                     /note="Ehrlichia chaffeensis str. Arkansas genomic region"
     regulatory      1334..1706
                     /gene="tuf-2"
                     /regulatory_class="promoter"
     gene            1334..1706
                     /gene="tuf-2"
                     /label=tuf-2
     CDS             1707..2495
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside-3'-adenylyltransferase"
                     /label=aadA
                     /protein_id="ATY38622.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            1707..2495
                     /gene="aadA"
                     /label=aadA
     misc_feature    2502..3536
                     /label=Ehrlichia chaffeensis str. Arkansas genomic region
                     /note="Ehrlichia chaffeensis str. Arkansas genomic region"
     promoter        complement(3607..3625)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3632..3648)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3789..4217
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             4561..5352
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             5373..6230
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6404..6992
                     /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 cctttatggg
      301 cctaagatag tattaccaat ttgtgcagct ctaacgttta ttggattagt gccattgcta
      361 atatcagatg tgtggttatt tgctcaaatt gggagaataa ttacaggtgt aggttcagct
      421 ggttctgctc ttggcctttt taaagttgct agtatgtact atggcaacag atttgctaga
      481 atgtctggta tttctgttat aataggactg ctaggtgcaa tgtatggagg actgccaatt
      541 ttatctttgt taaataaatt tggttgggaa agtttattct tagtatttat tattatagga
      601 gctgttatag cgctgttgct atatttgttt atgttgcctt atgataaaga ctccaatgtt
      661 gaagataata aaggtctctg tgataaaatt aagttgattg tatttaataa gtacattgtt
      721 atgatcagct tacttgcagg ctttatgatt ggacctttgg aaggttttgc tgatggatgg
      781 gttacttcat ttttaaaggc agttagtaat atggataaag aagtagctgc tttattgcct
      841 tctacaatat ttattggtat gtgctttgga ttgtttgtct taccatatat gttggagaag
      901 aaatcattta atagttggaa tatacttatc acatctgctt tggggatgtt gtttcctttc
      961 ctgatgttat atatcagtag ttcggttata ttggtcacaa tatcattttt tatgataggg
     1021 tttttttcat catatcagat tatagcaact tgtaaggtac taagttatgt tagtaataat
     1081 gtagttgcat tagctactgc tgtaaataac atgatagcta tggcttttgg ttatttcttt
     1141 catactgcta tatcttgtgt aatagatttg ttatgggatg gaaaaattgt agattcagag
     1201 ccagtatata ctaaggcatt aatgctaaaa tctatgttat ttattcctgg tggattgctt
     1261 ataggagcaa tagggtttat ttacctgaaa tatttggata agaaagaggg taagtaacac
     1321 atggtactac agtctcgaga aaaatgtgac tattaatttt gacttttttt aattaggtat
     1381 tgacatattg taatctttaa tttagttaaa tcaattctaa tctaaatttt tttggtatca
     1441 ttttttgttt ttttgtgtta attaatacaa ggatgttagg ttagcgttag tgtttttagg
     1501 aggagtggcc gagcggtcaa aggcagcaga ctgtaaatct gcccactagc gtgtacgtag
     1561 gttcaaatcc tacctcctcc acgggagatt atgcgggtat agctcaatgg tagagtccca
     1621 gccttccaag ctggctacgt gggttcgatt cccactatcc gctctttatt tttgaaatgg
     1681 tttaatgatg ttaaaggtat ttgtttatga gggaagcggt gatcgccgaa gtatcgactc
     1741 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     1801 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     1861 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     1921 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     1981 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     2041 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     2101 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     2161 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     2221 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     2281 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     2341 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctagg caggcttatc
     2401 ttggacaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gttcactacg
     2461 tgaaaggcga gatcaccaag gtagtcggca aataactcga gtatatataa tcatgtatcg
     2521 attatatata actgtgtgct ttgataaaac gatattagtt tgtatagttt tttgtgattt
     2581 aaaacgtcac tctaagttta agtatagagt ttgttcttac ttatgtgttc taagtagaaa
     2641 agctaatatc tacaacaaat agtttttttc ttcttatttt tagtaatcaa taatttaata
     2701 taagctatgc gactttttat atactttgat tttaaagtat tagaacatag attgtaactt
     2761 taaagttaag ataataaagc tctgttccag tttttgctat tgttaataat ctgatgattt
     2821 caattaaagt tgtattagtt tttgactgtt tgagatgtga atttagtagg ataatttaaa
     2881 tttacagcag taaagttata tattttgttt ctagtgttta tcagataatt tcataatggt
     2941 gtatcatgag ttctatgtat attttaatat aatatacttt acaacaaaac tttttgtaaa
     3001 ttctaagata ggtatgtcag attatacaga ttgaatagta ctgagatacc attttttatt
     3061 tgaaagtgtt gtatctttcc taaaggtcca tgtgtcttca atcttgttaa tttttgaagt
     3121 actaccctgt aggatattat cgtccttgtc ttttacaaag tttatttgtt cgctaatgaa
     3181 ttttactgtt atgtatgcta tattaccaga aagacttaac tttgttattt tgtgggatgt
     3241 tattgataca ataatatctt catatagttg accaagtgct ttacgatttt ctatttcttc
     3301 tacgaatgaa ttatataaat tagtatctaa tagcatagat aatatgtgta tatttccttc
     3361 attgaaagct tttattataa tttcaaatgc ctttattgac ccatccatga aatgtcttag
     3421 agaaaattct ttgtctttct ttttaatatg gttgatgatt gatttatcat tttcgttgca
     3481 aatttccatt acggaatcta tgtcaatatc ggcagtgtca tgttcttgtg tgtcttaagg
     3541 gcgaattctg cagatatcca tcacactggc ggccgctcga gcatgcatct agagggccca
     3601 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     3661 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3721 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3781 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3841 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3901 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3961 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4021 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4081 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4141 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4201 aaaatttaac gcgaatttta acaaaattca gggcgcaagg gctgctaaag gaagcggaac
     4261 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
     4321 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
     4381 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     4441 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     4501 aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc
     4561 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4621 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4681 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4741 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4801 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4861 gatctcctgt catcccacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4921 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4981 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5041 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5101 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5161 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5221 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5281 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5341 gacgagttct tctgaattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5401 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5461 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5521 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5581 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5641 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5701 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5761 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5821 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5881 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5941 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6001 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6061 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6121 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6181 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6241 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6301 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6361 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6421 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6481 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6541 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6601 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6661 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6721 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6781 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6841 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6901 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6961 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7021 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7081 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7141 agcgcagcga gtcagtgagc gaggaagcgg aag
//
