LOCUS       sequence-252669-        3016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kanamycin resistance donor for ShCAST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3016)
  AUTHORS   Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin
            EV, Zhang F
  TITLE     RNA-guided DNA insertion with CRISPR-associated transposases.
  JOURNAL   Science. 2019 Jul 5;365(6448):48-53. doi: 10.1126/science.aax9181. 
            Epub 2019 Jun 6.
  PUBMED    31171706
REFERENCE   2  (bases 1 to 3016)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.aax9181"; journalName: "Science"; date: "2019-07-5-
            5"; volume: "365"; issue: "6448"; pages: "48-53"
COMMENT     Sequence Label: pDonor_ShCAST_kanR
FEATURES             Location/Qualifiers
     source          1..3016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..48)
                     /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)."
     CDS             435..1229
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(489..508)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1099..1118
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(1337..1384)
                     /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)."
     terminator      complement(1432..1518)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1621..1715)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2059..2415
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            2466..2574
                     /note="incP origin of transfer"
ORIGIN
        1 gaagttccta tactttctag agaataggaa cttcggaata ggaacttcaa gatcccctga
       61 ttccctttgt caacagcaat ggataattcg atttaacaaa tgcatggcgc aagggctgct
      121 aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
      181 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
      241 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
      301 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
      361 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat
      421 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
      481 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
      541 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
      601 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
      661 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
      721 aagtgccggg gcaggatctc ctgtcatccc accttgctcc tgccgagaaa gtatccatca
      781 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
      841 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
      901 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
      961 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     1021 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     1081 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     1141 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     1201 tctatcgcct tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga ggatccaaca
     1261 tttccaatca ctagtgaatt atctagaatt attccattga gtaagttttt aagcacatca
     1321 gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggta
     1381 cttcaagatc cccaattcga gatcgtccgg gccgcaagct cctagcggcg gatttgtcct
     1441 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
     1501 agcctttcgt tttatttgat gcctcaagct agagagtcat taccccaggc gtttaagggc
     1561 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tctagcttgg
     1621 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     1681 ttctgaggtc attactggat ctatcaacag gagtccaagc tcagctaatt aaggcgacag
     1741 tcaatttgtc attatgaaaa tacacaaaag ctttttccta tcttgcaaag cgacagctaa
     1801 tttgtcacaa tcacggacaa cgacatctat tttgtcactg caaagaggtt atgctaaaac
     1861 tgccaaagcg ctataatcta tactgtataa ggattttact gatgacaata atttgtcaca
     1921 acgacatata attagtcact gtacacgtag agacgtagca atgctacctc tctacaatgg
     1981 ttttgtatgg tgcactctca gtacaatctt gtagcaccgc ctacatacct cgctctgcta
     2041 atcctgttac cagccggttg tcagccgtta agtgttcctg tgtcactcaa aattgctttg
     2101 agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac aaccgttaaa
     2161 ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt aaaaccttag
     2221 aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga gcttagtacg
     2281 tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc atgagggttt
     2341 agttcgttaa acatgagagc ttagtacgtt aaacttgaga gcttagtacg tgaaacatga
     2401 gagcttagta cgtactatca acaggttgaa ctgcccatgt tctttcctgc gttatcagag
     2461 cttatcggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     2521 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     2581 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     2641 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     2701 gcggaaagta aaaaattttt agtttattag acatctccac aaaaggcgta gtgtacagtg
     2761 acaaattatc tgtcgtcggt gacagattaa tgtcattgtg actatttaat tgtcgtcgtg
     2821 acccatcagc gttgcttaat taattgatga caaattaaat gtcatcaata taatatgctc
     2881 tgcaattatt atacaaagca attaaaacaa gcggataaaa ggacttgctt tcaacccacc
     2941 cctaagttta atagttactg agggggatcc actagtgagc tcatgcatga tctcgaatta
     3001 gcttcaaaag cgctct
//
