LOCUS       sequence-306811-        4162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicidal integration plasmid with the Tn7 arms, streptomycin and 
            kanamycin resistance genes and the restriction site to clone the 
            barcode-carrying cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4162)
  AUTHORS   Jasinska W, Manhart M, Lerner J, Gauthier L, Serohijos AWR, 
            Bershtein S
  TITLE     Chromosomal barcoding of E. coli populations reveals lineage 
            diversity dynamics at high resolution.
  JOURNAL   Nat Ecol Evol. 2020 Feb 24. pii: 10.1038/s41559-020-1103-z. doi: 
            10.1038/s41559-020-1103-z.
  PUBMED    32094541
REFERENCE   2  (bases 1 to 4162)
  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"; journalName: "Nat Ecol 
            Evol. 2020 Feb 24. pii: 10.1038/s41559-020-1103-z. doi: 
            10.1038/s41559-020-1103-z."
COMMENT     Sequence Label: Tn7 integration plasmid
FEATURES             Location/Qualifiers
     source          1..4162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    20..67
                     /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(145..172)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(264..350)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             953..1744
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     mobile_element  1850..2015
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2199..2587
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      complement(2648..2894)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    2974..3007
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             join(3375..4162,1..7)
                     /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(3429..3448)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4039..4058
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cttctaataa ggggatcttg aagttcctat tccgaagttc ctattctcta gaaagtatag
       61 gaacttcgaa gcagctccag cctacacaat cgctcaagac gtgtaatgct gcaatctgca
      121 tgcaagcttg gcactggcca cgcaaaaagg ccatccgtca ggatggcctt ctgcttaatt
      181 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcgca
      241 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
      301 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca
      361 gttccctact ctcgcatggg gagaccccac actaccatcg gggggcccag atcctctacg
      421 ccggacgcat cgtggcctgt gggcggacaa taaagtctta aactgaacaa aatagatcta
      481 aactatgaca ataaagtctt aaactagaca gaatagttgt aaactgaaat cagtccagtt
      541 atgctgtgaa aaagcatact ggacttttgt tatggctaaa gcaaactctt cattttctga
      601 agtgcaaatt gcccgtcgta ttaaagaggg gcgtggggtc gacgcggccg tggcgcgcct
      661 cctaggccgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
      721 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
      781 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
      841 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
      901 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatgaggga
      961 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     1021 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     1081 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     1141 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     1201 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     1261 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     1321 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     1381 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     1441 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     1501 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     1561 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     1621 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     1681 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     1741 ataactcgag tggcggccgc tattgaggga tctgattaat taaaacgtgg cttactagga
     1801 tccgatatca tttaaatcta gggataacag ggtaatacta gtgtcgacca accagataag
     1861 tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac
     1921 acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca
     1981 cccctcccag ttcccaacta ttttgtccgc ccacaggctg aaagcgctat ttcttccaga
     2041 attgccatga ttttttcccc acgggaggcg tcactggctc ccgtgttgtc ggcctgtcag
     2101 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttttgcggc
     2161 cggccgcaag atccggccac gatgcgtccg gcgtagagga tctgaagatc agcagttcaa
     2221 cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     2281 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
     2341 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
     2401 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
     2461 tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac
     2521 gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg
     2581 gctgacatgg gaattagcca tggcaaagcc tcgcaatcca gtgcaaagct agcttcttcg
     2641 tctgtttcta ctggtattgg cacaaacctg attccaattt gagcaaggct atgtgccatc
     2701 tcgatactcg ttcttaactc aacagaagat gctttgtgca tacagcccct cgtttattat
     2761 ttatctcctc agccagccgc tgtgctttca gtggatttcg gataacagaa aggccgggaa
     2821 atacccagcc tcgctttgta acggagtaga gacgaaagtg attgcgccta cccggatatt
     2881 atcgtgagga tgcgtcatcg ccattaattc actgatcagt gataagctgt caaacatgag
     2941 aattaattcc ggggatccgt cgacctgcag ttcgaagttc ctattctcta gaaagtatag
     3001 gaacttcaga gcgcttttga agctcacgct gccgcaagca ctcagggcgc aagggctgct
     3061 aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
     3121 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
     3181 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
     3241 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
     3301 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat
     3361 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3421 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3481 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3541 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3601 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3661 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3721 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3781 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3841 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3901 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3961 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4021 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4081 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4141 tctatcgcct tcttgacgag tt
//
