LOCUS       EZTN5KMSCE              1239 bp    DNA     linear   SYN 01-NOV-2020
DEFINITION  Synthetic transposon EZ-Tn5 <KAN-I-SceI>, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    Sce-poson
SOURCE      synthetic transposon EZ-Tn5 <KAN-I-SceI>
  ORGANISM  synthetic transposon EZ-Tn5 <KAN-I-SceI>
            other sequences; artificial sequences; vectors.
REFERENCE   1 (Mosaic Ends)
  AUTHORS   Zhou,M., Bhasin,A. and Reznikoff,W.S.
  TITLE     Molecular genetic analysis of transposase–end DNA sequence 
            recognition: cooperativity of three adjacent base-pairs in specific 
            interaction with a mutant Tn5 transposase
  JOURNAL   J. Mol. Biol. 276, 913–925 (1998)
   PUBMED   9566196
REFERENCE   2 (Description of EZ-Tn5)
  AUTHORS   Goryshin,I.Y., Jendrisak,J., Hoffman,L.M., Meis,R. and 
            Reznikoff,W.S.
  TITLE     Insertional transposon mutagenesis by electroporation of released 
            Tn5 transposition complexes
  JOURNAL   Nat. Biotechnol. 18(1), 97-100 (2000)
   PUBMED   10625401
REFERENCE   3 (Sce-poson construction)
  AUTHORS   Kang,Y., Durfee,T., Glasner,J.D., Qiu,Y., Frisch,D., Winterberg,K.M. 
            and Blattner,F.R.
  TITLE     Systematic mutagenesis of the Escherichia coli genome
  JOURNAL   J. Bacteriol. 186(15), 4921–4930 (2004)
   PUBMED   15262929
COMMENT     EZ-Tn5 is a trademark of Lucigen Corporation. The Sce-poson was 
            created by PCR amplification of EZ-Tn5 <KAN-2> with appropriate 
            primers to introduce the 18-bp I-SceI recognition site downstream 
            of the kanamycin resistance gene.
FEATURES             Location/Qualifiers
     source          1..1239
                     /organism="synthetic transposon EZ-Tn5 <KAN-I-SceI>"
                     /mol_type="other DNA"
     repeat_region   order(1..19,1221..1239)
                     /note="ME (hyperactive Mosaic End)"
                     /rpt_type=inverted
     primer_bind     complement(43..57)
                     /note="KAN-2 RP-1 Reverse Primer"
     gene            138..953
                     /gene="nptI"
     CDS             138..953
                     /note="kanamycin resistance gene"
                     /gene="nptI"
                     /gene_synonym="aph(3')-Ia"
                     /gene_synonym="neo"
                     /gene_synonym="aphA1"
                     /codon_start=1
                     /transl_table=11
                     /product="Aminoglycoside 3'-phosphotransferase"
                     /EC_number="2.7.1.95"
                     /note="APH(3')-I; Kanamycin kinase, type I;
                     Neomycin-kanamycin phosphotransferase type I; APH(3')I"
                     /function="confers resistance to neomycin, kanamycin, 
                     paromomycin, neamine, paromamine, vistamycin, gentamicin A,
                     and G418 (Geneticin(R))"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"                     
     primer_bind     1127..1151
                     /note="KAN-2 FP-1 Forward Primer"
     misc_feature    1190..1207
                     /note="I-SceI recognition site"
ORIGIN      
        1 CTGTCTCTTA TACACATCTC AACCATCATC GATGAATTGT GTCTCAAAAT CTCTGATGTT
       61 ACATTGCACA AGATAAAAAT ATATCATCAT GAACAATAAA ACTGTCTGCT TACATAAACA
      121 GTAATACAAG GGGTGTTATG AGCCATATTC AACGGGAAAC GTCTTGCTCG AGGCCGCGAT
      181 TAAATTCCAA CATGGATGCT GATTTATATG GGTATAAATG GGCTCGCGAT AATGTCGGGC
      241 AATCAGGTGC GACAATCTAT CGATTGTATG GGAAGCCCGA TGCGCCAGAG TTGTTTCTGA
      301 AACATGGCAA AGGTAGCGTT GCCAATGATG TTACAGATGA GATGGTCAGA CTAAACTGGC
      361 TGACGGAATT TATGCCTCTT CCGACCATCA AGCATTTTAT CCGTACTCCT GATGATGCAT
      421 GGTTACTCAC CACTGCGATC CCCGGAAAAA CAGCATTCCA GGTATTAGAA GAATATCCTG
      481 ATTCAGGTGA AAATATTGTT GATGCGCTGG CAGTGTTCCT GCGCCGGTTG CATTCGATTC
      541 CTGTTTGTAA TTGTCCTTTT AACAGCGATC GCGTATTTCG TCTCGCTCAG GCGCAATCAC
      601 GAATGAATAA CGGTTTGGTT GATGCGAGTG ATTTTGATGA CGAGCGTAAT GGCTGGCCTG
      661 TTGAACAAGT CTGGAAAGAA ATGCATAAAC TTTTGCCATT CTCACCGGAT TCAGTCGTCA
      721 CTCATGGTGA TTTCTCACTT GATAACCTTA TTTTTGACGA GGGGAAATTA ATAGGTTGTA
      781 TTGATGTTGG ACGAGTCGGA ATCGCAGACC GATACCAGGA TCTTGCCATC CTATGGAACT
      841 GCCTCGGTGA GTTTTCTCCT TCATTACAGA AACGGCTTTT TCAAAAATAT GGTATTGATA
      901 ATCCTGATAT GAATAAATTG CAGTTTCATT TGATGCTCGA TGAGTTTTTC TAATCAGAAT
      961 TGGTTAATTG GTTGTAACAC TGGCAGAGCA TTACGCTGAC TTGACGGGAC GGCGGCTTTG
     1021 TTGAATAAAT CGAACTTTTG CTGAGTTGAG GGATCAGATC ACGCATCTTC CCGACAACGC
     1081 AGACCGTTCC GTGGCAAAGC AAAAGTTCAA AATCACCAAC TGGTCCACCT ACAACAAAGC
     1141 TCTCATCAAC CGTGGCGGGG ATCCTCTAGA GTCGACCTGC AGGCATGCAT AGGGATAACA
     1201 GGGTAATAGC TTCAGGGTTG AGATGTGTAT AAGAGACAG
//