LOCUS       sequence-469016-        6690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose-inducible Tn5 InducTn-seq plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6690)
  AUTHORS   Basta DW, Campbell IW, Sullivan EJ, Hotinger JA, Hullahalli K, Garg 
            M, Waldor MK
  TITLE     Inducible transposon mutagenesis identifies bacterial fitness 
            determinants during infection in mice.
  JOURNAL   Nat Microbiol. 2025 Mar 27. doi: 10.1038/s41564-025-01975-z.
  PUBMED    40148565
REFERENCE   2  (bases 1 to 6690)
  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 
            Microbiol. 2025 Mar 27. doi: 10.1038/s41564-025-01975-z."
COMMENT     Sequence Label: pTn Donor
FEATURES             Location/Qualifiers
     source          1..6690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(34..849)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(34..849)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     757..776
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(850..941)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    974..992
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     promoter        1012..1296
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1219..1238
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    2866..2899
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  3142..3307
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        3441..3545
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3546..3614,3615..4406)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3764..3783)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4473..4861
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            4907..5016
                     /note="incP origin of transfer"
     primer_bind     5559..5575
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5560..5576
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5664..6197)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    6217..6250
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(6296..6382)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(6485..6579)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    6623..6641
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
ORIGIN
        1 gtagatctcg gtggtcgccg tatcattact agtttagaaa aactcatcga gcatcaaatg
       61 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
      121 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
      181 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
      241 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
      301 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
      361 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
      421 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
      481 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
      541 accggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
      601 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
      661 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
      721 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
      781 taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt cccgttgaat
      841 atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      901 gagcggatac atatttgaat gtatttagaa aaataaacaa aagagtttga cggatggcct
      961 ttttgcgttt ctaagatgtg tataagagac agatactccc gccattcaga gaagaaacca
     1021 attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa
     1081 ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga
     1141 caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc
     1201 acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg gatcctacct
     1261 gacgcttttt atcgcaactc tctactgttt ctccataccc gacttataac tctactctct
     1321 taggactaat atttatgata acttctgctc ttcatcgtgc ggccgactgg gctaaatctg
     1381 tgttctcttc ggcggcgctg ggtgatcctc gccgtactgc ccgcttggtt aacgtcgccg
     1441 cccaattggc aaaatattct ggtaaatcaa tacccatctc atcagagggt agtgaagccg
     1501 cccaggaagg cgcttaccga tttatccgca atcccaacgt ttctgccgag gcgatcagaa
     1561 aggctggcgc catgcaaaca gtcaagttgg ctcaggagtt tcccgaactg ctggccattg
     1621 aggacaccac ctctttgagt tatcgccacc aggtcgccga agagcttggc aagctgggct
     1681 ctattcagga taaatcccgc ggatggtggg ttcactccgt tctcttgctc gaggccacca
     1741 cattccgcac cgtaggatta ctgcatcagg agtggtggat gcgcccggat gaccctgccg
     1801 atgcggatga aaaggagagt ggcaaatggc tggcagcggc cgcaactagc cggttacgca
     1861 tgggcagcat gatgagcaac gtgattgcgg tctgtgaccg cgaagccgat attcatgctt
     1921 atctgcagga caaactggcg cataacgagc gcttcgtggt gcgctccaag cacccacgca
     1981 aggacgtaga gtctgggttg tatctgtacg accatctgaa gaaccaaccg gagttgggtg
     2041 gctatcagat cagcattccg caaaagggcg tggtggataa acgcggtaaa cgtaaaaatc
     2101 gaccagcccg caaggcgagc ttgagcctgc gcagtgggcg catcacgcta aaacagggga
     2161 atatcacgct caacgcggtg ctggccgagg agattaaccc gcccaagggt gagaccccgt
     2221 tgaaatggtt gttgctgacc agcgaaccgg tcgagtcgct agcccaagcc ttgcgcgtca
     2281 tcgacattta tacccatcgc tggcggatcg aggagttcca taaggcatgg aaaaccggag
     2341 caggagccga gaggcaacgc atggaggagc cggataatct ggagcggatg gtctcgatcc
     2401 tctcgtttgt tgcggtcagg ctgttacagc tcagagaaag cttcacgctg ccgcaagcac
     2461 tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg
     2521 tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca
     2581 aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta
     2641 tggacagcaa gcgaaccgga attgccagct ggggcgccct ctgggaaggt tgggaagccc
     2701 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga
     2761 tctgatcggt cgacctgcag atctgcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2821 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcataac ttcgtataat
     2881 gtatgctata cgaacggtag ctgatgtgct cagtatcacc gccagtggta tttatgtcaa
     2941 caccgccaga gataatttat caccgcagat ggttatctgt atgtttttta tatgaattta
     3001 ttttttgcag gggggcattg tttggtaggt gagagatctg aattgctatg tttagtgagt
     3061 tgtatctatt tatttttcaa taaatacaat tggttacctc gagaagcttg ggcccggtac
     3121 ctcgcgaagg ccttgcaggc caaccagata agtgaaatct agttccaaac tattttgtca
     3181 tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct
     3241 tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc
     3301 gcccacagcg gggcattttt cttcctgtta tgtttgggcg ctgcattaat gaatcggggc
     3361 catatcaatg attttctggt gcgtaccggg ttgagaagcg gtgtaagtga actgcaggtg
     3421 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3481 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3541 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3601 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3661 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3721 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3781 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3841 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3901 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3961 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4021 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4081 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4141 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4201 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4261 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4321 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4381 gtgcctcact gattaagcat tggtaatttt tttaaggcag ttattggtgc ctagaaatat
     4441 tttatctgat taataagatg atcttcgtag aggatctgaa gatcagcagt tcaacctgtt
     4501 gatagtacgt actaagctct catgtttcac gtactaagct ctcatgttta acgtactaag
     4561 ctctcatgtt taacgaacta aaccctcatg gctaacgtac taagctctca tggctaacgt
     4621 actaagctct catgtttcac gtactaagct ctcatgtttg aacaataaaa ttaatataaa
     4681 tcagcaactt aaatagcctc taaggtttta agttttataa gaaaaaaaag aatatataag
     4741 gcttttaaag cttttaaggt ttaacggttg tggacaacaa gccagggatg taacgcactg
     4801 agaagccctt agagcctctc aaagcaattt tgagtgacac aggaacactt aacggctgac
     4861 atgggaattg taccccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc
     4921 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
     4981 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa
     5041 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat
     5101 accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa gcgctgcttc
     5161 cctgctgttt tgtggaatat ctaccgactg gaaacaggca aatgcaggaa attactgaac
     5221 tgaggggaca ggcgagagac gatgccaaag agctacaccg acgagctggc cgagtgggtt
     5281 gaactagagg accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat
     5341 gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa
     5401 actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta
     5461 tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa
     5521 gtgcaaattg cccgtcgtat taaagagggg cgtggggttc gaggtcgacg gtatcgataa
     5581 gctagatcga attggccgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
     5641 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
     5701 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
     5761 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
     5821 agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag
     5881 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc
     5941 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
     6001 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa
     6061 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg
     6121 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg
     6181 ttgctgctgc gtaacatcgt tgctgctcca taacattacc gttcgtataa tgtatgctat
     6241 acgaagttat aagatcccca attcgatcgt ccgggccgca agctcctagc ggcggatttg
     6301 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
     6361 actgagcctt tcgttttatt tgatgcctca agctagagag tcattacccc aggcgtttaa
     6421 gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtctagc
     6481 ttggattctc accaataaaa aacgcctggc ggcaaccgag cgttctgaac aaatccagat
     6541 ggagttctga ggtcattact ggatctatca acaggagtcc aagctcagct aattaagcta
     6601 gcttatcgat accgttaatt aactgtctct tatacacatc tatttctaga tcggaagagc
     6661 gtcgtgtagg gaaagagtgt agatctcggt
//
