LOCUS       sequence-399455-        3454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZooTsy donor harboring KanR cargo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3454)
  AUTHORS   Faure G, Saito M, Benler S, Peng I, Wolf YI, Strecker J, Altae-Tran 
            H, Neumann E, Li D, Makarova KS, Macrae RK, Koonin EV, Zhang F
  TITLE     Modularity and diversity of target selectors in Tn7 transposons.
  JOURNAL   Mol Cell. 2023 Jun 15;83(12):2122-2136.e10. doi: 
            10.1016/j.molcel.2023.05.013. Epub 2023 Jun 1.
  PUBMED    37267947
REFERENCE   2  (bases 1 to 3454)
  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: 
            "10.1016/j.molcel.2023.05.013"; journalName: "Mol Cell"; date: 
            "2023-06-15- 15"; volume: "83"; issue: "12"; pages: "2122-2136.e10"
COMMENT     Sequence Label: pSCT384: pSC101-Donor(ZooTsy)
FEATURES             Location/Qualifiers
     source          1..3454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(449..496)
                     /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             883..1677
                     /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(937..956)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1547..1566
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(1901..2851)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2899..3121
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt
       61 ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag
      121 taaggcagcg gtatcatcaa cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      181 gattttggtc atgagattat caaaaaggat cttcacctag atccgttatg cagcggaaag
      241 tagacgagct gccgaagttc tgatttagcc aatgtagcaa cgaacaaggc aaaaattcgg
      301 ataacccact gactataaag gaatcccaaa gtatccaatg tcgcattact tgcaggaatc
      361 ctgtgagtga tgtcacagca ttagaagtag tcggggggat ccactagtga gctcatgcat
      421 gatctcgaat tagcttcaaa agcgctctga agttcctata ctttctagag aataggaact
      481 tcggaatagg aacttcaaga tcccctgatt ccctttgtca acagcaatgg ataattcgat
      541 ttaacaaatg catggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg
      601 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
      661 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg
      721 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt
      781 gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg
      841 ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
      901 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
      961 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     1021 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     1081 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     1141 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatcccac
     1201 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     1261 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     1321 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     1381 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     1441 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     1501 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     1561 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     1621 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt
     1681 gaaaaaggaa gagtatgagg atccaacatt tccaatcact agtgaattat ctagaattat
     1741 tccattgagt aagtttttaa gcacatcagc ttcaaaagcg ctctttattg gaagcgctaa
     1801 ggaatattgg agaatcccag tccgaattcg accgacctct ctacaatggt tttgtgttag
     1861 tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag
     1921 ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga
     1981 gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt
     2041 ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg
     2101 taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta
     2161 cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc
     2221 gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca
     2281 aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact
     2341 taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt
     2401 ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca
     2461 gattatattt tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa
     2521 ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc
     2581 ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag
     2641 ctaatacacc ataagcattt tccctactga tgttcatcat ctgaacgtat tggttataag
     2701 tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca
     2761 cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt
     2821 catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat
     2881 cactatacca attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg
     2941 tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct
     3001 ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt
     3061 atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact
     3121 cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc
     3181 tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc tcggttgcgg
     3241 ccgcaatcgg gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg
     3301 ctgtcttttt cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa
     3361 atggcactac aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa
     3421 gcccgtcacg ggcttctcag ggcgttttat ggcg
//
