LOCUS       40924_20081        4154 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Kanamycin resistance FRT vector pFKM2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4154)
  AUTHORS   Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer 
            HP.
  TITLE     Genetic tools for select-agent-compliant manipulation of 
            Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 74 (4), 1064-1075 (2008)
  PUBMED    18156318
REFERENCE   2  (bases 1 to 4154)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Genetic tools for Pseudomonas and its related bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4154)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-SEP-2007) Microbiology, Immunology and Pathology, 
            Colorado State University, 1682 Campus Delivery, Ft. Collins, CO 
            80523-1682, USA
REFERENCE   4  (bases 1 to 4154)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4154)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "4"; pages:
            "1064-1075"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (21-SEP-2007) Microbiology, Immunology and Pathology, Colorado State
            University, 1682 Campus Delivery, Ft. Collins, CO 80523-1682, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(401..457)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    466..513
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(623..1414)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    1800..1847
                     /note="FRT; Flp recombinase target site"
     protein_bind    1800..1847
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    1864..1920
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1933..1949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1957..1973)
                     /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)."
     promoter        complement(1981..2011)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2026..2047)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2335..2923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3097..3954)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3955..4059)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
      241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat gcctgcaggt
      421 cgactctaga ggatccccgg gtaccgagct cgaattgggg atcttgaagt acctattccg
      481 aagttcctat tctctagaaa gtataggaac ttcagagcgc ttttgaagct gatgtgctta
      541 aaaacttact caatggaata attctagata attcactagt gatggaaatg ttggatcctc
      601 atactcttcc tttttcaatt cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
      661 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
      721 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
      781 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
      841 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
      901 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
      961 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     1021 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     1081 tctcggcagg agcaaggtgg gatgacagga gatcctgccc cggcacttcg cccaatagca
     1141 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     1201 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     1261 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     1321 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     1381 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     1441 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     1501 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg
     1561 ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct
     1621 ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     1681 gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc
     1741 gccatgcatt tgttaatcga attatccatt gctgttgaca aagggaatca ggggatcttg
     1801 aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcaga gcgcttttga
     1861 agctaattcg agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt
     1921 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1981 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2041 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2161 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2221 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2281 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2761 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2941 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3001 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3061 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3121 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3181 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3241 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3301 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3361 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3421 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3481 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3541 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3601 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3661 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3721 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3781 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3841 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3901 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3961 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4021 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4081 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4141 gaggcccttt cgtc
//
