LOCUS       sequence-402567-        3619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AjTn6022 donor harboring KanR cargo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3619)
  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 3619)
  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: pSCT368: pSC101-Donor(AjTn6022)
FEATURES             Location/Qualifiers
     source          1..3619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(455..502)
                     /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             889..1683
                     /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(943..962)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1553..1572
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(2066..3016)
                     /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      3064..3286
                     /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 tatgtcattt acagcaatag ataagagtgt catgataaga aaacaatagg ttgccactct
      301 aaaattacac aaatagatta gagtaatgac agtaataagc aagagtggct tgcaaattta
      361 cacaaatacg cacgagtggc tgtaagaggc attctgtcgg ggggatccac tagtgagctc
      421 atgcatgatc tcgaattagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata
      481 ggaacttcgg aataggaact tcaagatccc ctgattccct ttgtcaacag caatggataa
      541 ttcgatttaa caaatgcatg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc
      601 agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg
      661 gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta
      721 gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt
      781 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
      841 cgcaggggat caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      901 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      961 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     1021 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     1081 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     1141 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     1201 tcccaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     1261 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     1321 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     1381 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     1441 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     1501 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     1561 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     1621 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     1681 tgaattgaaa aaggaagagt atgaggatcc aacatttcca atcactagtg aattatctag
     1741 aattattcca ttgagtaagt ttttaagcac atcagcttca aaagcgctct tcactgccct
     1801 cattcctaag aaacccacca tatctataga cagttctgcg aatattatca tttgggaaac
     1861 aatcaatagt tgtgacttta acttttttat ttgcaggcat atcttttagt gaatttaatg
     1921 aaattcatat ataaatagta tagtttttat tatcactcta gtttatttgt gtaaactctg
     1981 atttatttgt ggaactgaca cctctctaca atggttttgt gttagtcttg atgcttcact
     2041 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
     2101 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt
     2161 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
     2221 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta
     2281 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     2341 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     2401 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     2461 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     2521 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     2581 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     2641 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     2701 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat
     2761 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     2821 cattttccct actgatgttc atcatctgaa cgtattggtt ataagtgaac gataccgtcc
     2881 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     2941 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     3001 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     3061 gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat
     3121 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     3181 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
     3241 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
     3301 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
     3361 aaaaccctaa aggcttaagt agcaccctcg caagctcggt tgcggccgca atcgggcaaa
     3421 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
     3481 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
     3541 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt
     3601 ctcagggcgt tttatggcg
//
