LOCUS       40924_2557        4128 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p34E-Km, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4128)
  AUTHORS   Shastri S, Spiewak HL, Sofoluwe A, Eidsvaag VA, Asghar AH, Pereira 
            T, Bull EH, Butt AT, Thomas MS.
  TITLE     An efficient system for the generation of marked genetic mutants in 
            members of the genus Burkholderia
  JOURNAL   Plasmid (2016) In press
  PUBMED    27825973
REFERENCE   2  (bases 1 to 4128)
  AUTHORS   Shastri S, Spiewak HL, Pereira T, Sofoluwe AO, Eidsvaag VA, Bull EH,
            Asghar AH, Thomas MS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 4128)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4128)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 
            (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-JUN-2016) Department of Infection, Immunity "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        786..890
                     /label=AmpR promoter
     CDS             891..1748
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1922..2510
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2762..2780
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2789..2845)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(2884..3675)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(3779..3807)
                     /label=aphA2 promoter
                     /note="aphA2 promoter"
                     /regulatory_class="promoter"
     misc_feature    4049..4105
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(4110..4128)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg
       61 tacaagccta attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa
      121 actgccgtca gagtcggttt ggttggacga accttctgag tttctggtaa cgccgttccg
      181 caccccggaa atggtcagcg aaccaatcag cagggtcatc gctagccaga tcctctacgc
      241 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
      301 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
      361 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcacca
      421 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
      481 gagtcgcata agggagagcg tcgatatggt gcactctcag tacaatctgc tctgatgccg
      541 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      601 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
      661 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
      721 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
      781 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      841 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      901 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
      961 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1021 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1081 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1141 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1201 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1261 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1321 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1381 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1441 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1501 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1561 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1621 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1681 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1741 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     1801 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     1861 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1921 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1981 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2041 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2101 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2161 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2221 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2281 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2341 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2401 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2461 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2521 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2581 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2641 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2701 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cttatcgaaa
     2761 ttaatacgac tcactatagg gagaccggaa gcttgcatgc ctgcaggtcg actctagagg
     2821 atccccgggt accgagctcg aattcgcgtc gaattaattc cgcgaacccc agagtcccgc
     2881 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     2941 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     3001 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     3061 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     3121 gacgagatcc tcgccgtcgg gcatccgcgc cttgagcctg gcgaacagtt cggctggcgc
     3181 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     3241 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     3301 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     3361 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     3421 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     3481 cgctgcctcg tcttggagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     3541 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     3601 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     3661 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct
     3721 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
     3781 cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca
     3841 gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt
     3901 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     3961 actggctttc tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca
     4021 gcgtgaagct agcggaatta ttcgacgcga attcgagctc ggtacccggg gatcctctag
     4081 agtcgacctg caggcatgca agcttgtatt ctatagtgtc acctaaat
//
