LOCUS       40924_20796        3506 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Zeocin resistance FRT vector pFZE1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3506)
  AUTHORS   Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     A Tn7-based broad-range bacterial cloning and expression system
  JOURNAL   Nat. Methods 2 (6), 443-448 (2005)
  PUBMED    15908923
REFERENCE   2  (bases 1 to 3506)
  AUTHORS   Choi K-H., Gaynor JB, White KG, Lopez C, Bosio CM, 
            Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-JUL-2005) Microbiology, Immunology and Pathology, 
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 3506)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3506)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2005"; volume: "2"; issue: "6"; pages: "443-448"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-JUL-2005) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(399..455)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    464..511
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(625..996)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1015..1062)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     misc_feature    1152..1199
                     /label=FRT, Flp recombinase target site
                     /note="FRT, Flp recombinase target site"
     protein_bind    1152..1199
                     /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    1216..1272
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1285..1301)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1309..1325)
                     /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(1333..1363)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1378..1399)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1687..2275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2449..3306)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3307..3411)
                     /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 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atccccgggt accgagctcg aattggggat cttgaagtac ctattccgaa
      481 gttcctattc tctagaaagt ataggaactt cagagcgctt ttgaagctga tgtgcttaaa
      541 aacttactca atggaataat tctagataat tcggcttatc tctgatgtta cattgcacaa
      601 gataaaaata tatcatcatg atcagtcctg ctcctcggcc acgaagtgca cgcagttgcc
      661 ggccgggtcg cgcagggcga actcccgccc ccacggctgc tcgccgatct cggtcatggc
      721 cggcccggag gcgtcccgga agttcgtgga cacgacctcc gaccactcgg cgtacagctc
      781 gtccaggccg cgcacccaca cccaggccag ggtgttgtcc ggcaccacct ggtcctggac
      841 cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa
      901 gtcccgggag aacccgagcc ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc
      961 ggtgagcacc ggaacggcac tggtcaactt ggccatggtt tagttcctca ccttgtcgta
     1021 ttatactatg ccgatatact atgccgatga ttaattgtca acacgtgctg atcatgacca
     1081 aaatccctta acgtgagtta cgcgtcgaag ccgaaatcca ttgctgttga caaagggaat
     1141 caggggatct tgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttca
     1201 gagcgctttt gaagctaatt cgagctcggt acccggggat cctctagagt cgacctgcag
     1261 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1321 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1381 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1441 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1501 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1561 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1621 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2101 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     2161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2221 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2281 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2341 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2401 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2461 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2521 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2581 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2641 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2701 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2761 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2821 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2881 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2941 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3001 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3061 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3121 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3181 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3241 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3301 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3361 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3421 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3481 taggcgtatc acgaggccct ttcgtc
//
