LOCUS       40924_20086        4146 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pFKm4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4146)
  AUTHORS   Kvitko BH, Bruckbauer S, Prucha J, McMillan I, Breland EJ, Lehman S,
            Mladinich K, Choi KH, Karkhoff-Schweizer R, Schweizer HP.
  TITLE     A simple method for construction of pir+ Enterobacterial hosts for 
            maintenance of R6K replicon plasmids
  JOURNAL   BMC Res Notes 5, 157 (2012)
  PUBMED    22433797
REFERENCE   2  (bases 1 to 4146)
  AUTHORS   Kvitko BH, Goodyear A, Propst KL, Dow SW, Schweizer HP.
  TITLE     Burkholderia pseudomallei Known Siderophores and Hemin Uptake Are 
            Dispensable for Lethal Murine Melioidosis
  JOURNAL   PLoS Negl Trop Dis 6 (6), E1715 (2012)
  PUBMED    22745846
REFERENCE   3  (bases 1 to 4146)
  AUTHORS   Kvitko BH, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-AUG-2012) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   4  (bases 1 to 4146)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4146)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC Res 
            Notes 5, 157 (2012)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "PLoS Negl 
            Trop Dis"; date: "2012"; volume: "6"; issue: "6"; pages: "E1715"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (17-AUG-2012) Microbiology Immunology and Pathology, Colorado State 
            University, IDRC at Foothills Campus, Campus Delivery 0922, Fort 
            Collins, CO 80523-0922, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4146
                     /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(615..1406)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_recomb     1792..1839
                     /label=FRT
                     /note="FRT"
     protein_bind    1792..1839
                     /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    1856..1912
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1925..1941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1949..1965)
                     /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(1973..2003)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2018..2039)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2327..2915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3089..3946)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3947..4051)
                     /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 attaattaag tgattggaaa tgttggatcc tcatactctt
      601 cctttttcaa ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
      661 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
      721 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
      781 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
      841 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
      901 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
      961 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     1021 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     1081 ggagcaaggt gggatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     1141 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     1201 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     1261 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     1321 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     1381 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     1441 cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag
     1501 ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat
     1561 aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt
     1621 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     1681 cgtttctgcg gactggcttt ctacgtgttc cgcttccttt agcagccctt gcgccatgca
     1741 tttgttaatc gaattatcca ttgctgttga caaagggaat caggggatct tgaagttcct
     1801 attccgaagt tcctattctc tagaaagtat aggaacttca gagcgctttt gaagctaatt
     1861 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat
     1921 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1981 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2041 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2101 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2161 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2221 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2281 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2341 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2401 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2461 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2521 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2581 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2641 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2701 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2761 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2821 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2881 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2941 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3001 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3061 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3121 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3181 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3241 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3301 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3361 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3421 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3481 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3541 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3601 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3661 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3721 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3781 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3841 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3901 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3961 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4021 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4081 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4141 ttcgtc
//
