LOCUS       sequence-266499-        4434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transposon mutagenesis plasmid and the cI repressor expression 
            plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4434)
  AUTHORS   Neil K, Allard N, Grenier F, Burrus V, Rodrigue S
  TITLE     Highly efficient gene transfer in the mouse gut microbiota is 
            enabled by the Incl(2) conjugative plasmid TP114.
  JOURNAL   Commun Biol. 2020 Sep 22;3(1):523. doi: 10.1038/s42003-020-01253-0.
  PUBMED    32963323
REFERENCE   2  (bases 1 to 4434)
  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: "Commun 
            Biol."; date: "2020-09-22"; volume: "3(1):523. doi"; pages: " 
            10.1038/s42003-020-01253-0"
COMMENT     Sequence Label: pFG036 + pFG051
FEATURES             Location/Qualifiers
     source          1..4434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    44..62
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     protein_bind    complement(134..181)
                     /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)."
     protein_bind    complement(1266..1299)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1350..1369)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(2024..2057)
                     /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)"
     misc_feature    2128..2146
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     RBS             2326..2337
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2344..3771
                     /codon_start=1
                     /product="transposase from the bacterial Tn5 transposon 
                     (Reznikoff, 1993)"
                     /label=Tn5 transposase
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     oriT            3916..4024
                     /note="incP origin of transfer"
     rep_origin      4043..4431
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ggcgcgccca gtctagaaga aatcctaggt aaagtggccg gccctgtctc ttatacacat
       61 ctgacgctgc cgacgaatag agaggtgtag atctcggtgg tcgccgtatc attgtgtagg
      121 ctggagctgc ttcgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt
      181 catttaaatg gcgcgcctta cgcatcgatg acagtcattc atctttctgc ccctccaaaa
      241 gcaaaaaccc gccgaagcgg gtttttacgt aaatcaggtg aaactgaccg actcgagagg
      301 aacttcaaga attctcattg gctggcacca agcagtttaa ctgcttcata cttcacgaat
      361 ttaatgagcg ccgccacctg atccgcacga gcgggcaaat aatcttcttc ttgcccaaga
      421 taagcccgct tggcattcaa caggatgggc tgatgttgag ctggcaagcg tgccattgcc
      481 caagtggcag caacatcctt tggcgcgatc ttgccggttg ctgcggtgta ccagatacga
      541 gacaaagtaa gcactacatt ccgctcatcg cctgcccaat ctggcggcga gttccatagc
      601 ttcagagtat cggccagtgc cttgaataga tcgctttctg ggactgagct gaagagatcc
      661 ttcgctgctg aacctgccaa gacgacgcta tgttgctttg cctttgttag cagaatcgcc
      721 aagtcagaat cggttgtggc gggctcgaag atgcccgcaa ggatgtcttt gcgctgccac
      781 tctccgaact gcagttcccg cctggccgga taacgccaag gtacgatgtc actgtgcacg
      841 acgatggtca cttccaaggc gcggagtgcc ttgttttggc caggggaagc tgaaacctcc
      901 aagagatcga cgagcagggc ttgccgcacg gcatcattga gcggtgcagc tacagtcacc
      961 agcaaatcaa tatcactgta cggtttcaat ccgccatcca gtgcggagcc gtacaaatgc
     1021 acggccagca acgtcgactc caagtggcgc ccgatgacgt tgagtgcttg tgatagttgc
     1081 accgaaatct cggcgggcac tttttcactc atgcgaaacg atcctcatcc tgtctcttga
     1141 tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
     1201 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgcg
     1261 catgcataac ttcgtatagc atacattata cgaagttatc cggatatacc accgttgata
     1321 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     1381 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     1441 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     1501 tacgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     1561 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     1621 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     1681 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     1741 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     1801 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     1861 atgccgtttg tgatggcttc catgtcggca gatgcttaat gaatacaaca gtactgcgat
     1921 gaggatccta gagatcaagc cttaacgaac taagaccccc gcaccgaaag gtccgggggt
     1981 tttttttgac cttaaaaaca taaccgagga gcagacagac gtcgaagttc ctatactttc
     2041 tagagaatag gaacttcgga ataggcccgg gaactaagga ggatattcat atggaccatg
     2101 gctaattccc atgtctcgtg ggctcggaga tgtgtataag agacagctgg tttaaaccaa
     2161 ttcagatctc tcacctacca aacaatgccc ccctgcaaaa aataaattca tataaaaaac
     2221 atacagataa ccatctgcgg tgataaatta tctctggcgg tgttgacata aataccactg
     2281 gcggttatac tgagcacatc agcaggacgc actgaccgaa ttcataaaga ggagaaaggt
     2341 accatgatta ccagcgcact gcatcgtgcg gcggattggg cgaaaagcgt gttttctagt
     2401 gctgcgctgg gtgatccgcg tcgtaccgcg cgtctggtga atgttgcggc gcaactggcc
     2461 aaatatagcg gcaaaagcat taccattagc agcgaaggca gcaaagccat gcaggaaggc
     2521 gcgtatcgtt ttattcgtaa tccgaacgtg agcgcggaag cgattcgtaa agcgggtgcc
     2581 atgcagaccg tgaaactggc ccaggaattt ccggaactgc tggcaattga agataccacc
     2641 tctctgagct atcgtcatca ggtggcggaa gaactgggca aactgggtag cattcaggat
     2701 aaaagccgtg gttggtgggt gcatagcgtg ctgctgctgg aagcgaccac ctttcgtacc
     2761 gtgggcctgc tgcatcaaga atggtggatg cgtccggatg atccggcgga tgcggatgaa
     2821 aaagaaagcg gcaaatggct ggccgctgct gcaacttcgc gtctgagaat gggcagcatg
     2881 atgagcaacg tgattgcggt gtgcgatcgt gaagcggata ttcatgcgta tctgcaagat
     2941 aaactggccc ataacgaacg ttttgtggtg cgtagcaaac atccgcgtaa agatgtggaa
     3001 agcggcctgt atctgtatga tcacctgaaa aaccagccgg aactgggcgg ctatcagatt
     3061 agcattccgc agaaaggcgt ggtggataaa cgtggcaaac gtaaaaaccg tccggcgcgt
     3121 aaagcgagcc tgagcctgcg tagcggccgt attaccctga aacagggcaa cattaccctg
     3181 aacgcggtgc tggccgaaga aattaatccg ccgaaaggcg aaaccccgct gaaatggctg
     3241 ctgctgacca gcgagccggt ggaaagtctg gcccaagcgc tgcgtgtgat tgatatttat
     3301 acccatcgtt ggcgcattga agaatttcac aaagcgtgga aaacgggtgc gggtgcggaa
     3361 cgtcagcgta tggaagaacc ggataacctg gaacgtatgg tgagcattct gagctttgtg
     3421 gcggtgcgtc tgctgcaact gcgtgaatct tttactccgc cgcaagcact gcgtgcgcag
     3481 ggcctgctga aagaagcgga acacgttgaa agccagagcg cggaaaccgt gctgaccccg
     3541 gatgaatgcc aactgctggg ctatctggat aaaggcaaac gcaaacgcaa agaaaaagcg
     3601 ggcagcctgc aatgggcgta tatggcgatt gcgcgtctgg gcggctttat ggatagcaaa
     3661 cgtaccggca ttgcgagctg gggtgcgctg tgggaaggtt gggaagcgct gcaaagcaaa
     3721 ctggatggct ttctggccgc gaaagacctg atggcgcagg gcattaaaat ctaaactagt
     3781 cttttccgct gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc
     3841 ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat
     3901 ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt
     3961 cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg
     4021 gccgtaggcc ggccggcgcg ccgatctgaa gatcagcagt tcaacctgtt gatagtacgt
     4081 actaagctct catgtttcac gtactaagct ctcatgttta acgtactaag ctctcatgtt
     4141 taacgaacta aaccctcatg gctaacgtac taagctctca tggctaacgt actaagctct
     4201 catgtttcac gtactaagct ctcatgtttg aacaataaaa ttaatataaa tcagcaactt
     4261 aaatagcctc taaggtttta agttttataa gaaaaaaaag aatatataag gcttttaaag
     4321 cctttaaggt ttaacggttg tggacaacaa gccagggatg taacgcactg agaagccctt
     4381 agagcctctc aaagcaattt tgagtgacac aggaacactt aacggctgac atgg
//
