LOCUS       sequence-267150-        4483 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 4483)
  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 4483)
  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..4483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3126..3146
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     3147..3166
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3147..3165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3173..3190
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     3244..3262
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3300..3322)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3422..3441
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(3580..4293)
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgaatt cggccagtct acatgtactc tttttgataa aaaattggag attcctttac
      901 aaatatgctc ttacgtgcta ttatttaagt gactatttaa aaggagttaa taaatatgcg
      961 gcaaggtatt cttaaataaa ctgtcaattt gatagcggga acaaataatt agatgtcctt
     1021 ttttaggagg gcttagtttt ttgtacccag tttaagaata cctttatcat gtgattctaa
     1081 agtatccaga gaatatctgt atgctttgta tacctatggt tatgcataaa aatcccagtg
     1141 ataaaagtat ttatcactgg gatttttatg cccttttggg tttttgaatg gaggaaatct
     1201 agatgaaaat cataaatatc ggtgtattag ctcacgttga tgcaggaaaa acaacattaa
     1261 ctgaatcact tttatataac tctggtgcaa ttactgaact tggttcagta gataaaggta
     1321 ctactcgtac tgataataca ttattagaac gtcaacgtgg aatcacaatt caaacaggta
     1381 tcacatcttt tcaatgggaa aatacaaaag taaatattat agatacacct ggacacatgg
     1441 atttccttgc agaagtatac cgtagtcttt cagtattaga tggtgctatt ttacttatca
     1501 gcgctaaaga tggagttcaa gctcaaactc gtatcttatt tcacgcatta cgtaaaatgg
     1561 gtattccaac aattttcttt ataaacaaaa ttgaccaaaa cggaattgat ttaagtacag
     1621 tttatcaaga tatcaaagaa aaactttctg ctgaaatcgt tattaaacaa aaagttgaat
     1681 tatacccaaa cgtttgcgta acaaatttta ctgaatcaga acaatgggat acagttatag
     1741 aaggtaatga tgatttatta gaaaaataca tgtcaggtaa atcattagaa gcattagaat
     1801 tagaacaaga agaaagtatt cgtttccaaa actgttcttt attcccttta taccatggaa
     1861 gcgctaaaag taacataggt attgataact taattgaagt tattactaac aaattttact
     1921 cttcaactca tcgtggtcct tctgaattat gcggtaacgt tttcaaaatt gaatatacaa
     1981 aaaaacgtca acgtttagct tatatacgtc tttatagtgg tgttttacat ttacgtgata
     2041 gtgttcgtgt tagtgaaaaa gaaaagatta aagttacaga aatgtatact tctattaacg
     2101 gtgaattatg caaaattgac cgtgcatatt caggtgaaat tgtaatttta caaaacgaat
     2161 ttcttaaact taatagtgta cttggtgaca caaaactttt accacaacgt aagaaaattg
     2221 aaaatccaca cccattactt caaacaacag tagaaccaag caaacctgaa caacgtgaaa
     2281 tgcttttaga tgctctttta gaaattagtg actctgaccc acttttacgt tactatgtag
     2341 attctactac tcatgaaatt attctttctt tccttggtaa agttcaaatg gaagttattt
     2401 ctgcattatt acaagaaaaa tatcatgttg aaatcgaatt aaaagaacct actgtaattt
     2461 atatggaacg tccattaaaa aatgctgaat atacaattca tattgaagtt ccaccaaatc
     2521 cattttgggc ttctattggt ctttctgttt ctccacttcc acttggtagc ggaatgcaat
     2581 atgaaagtag cgtaagttta ggttatctta atcaaagttt ccaaaacgca gttatggaag
     2641 gtattcgtta cggttgcgaa caaggtttat acggttggaa tgttacagac tgcaaaatct
     2701 gttttaagta tggactttac tattcacctg tatcaacacc tgctgacttt cgtatgcttg
     2761 caccaattgt tttagaacaa gttttaaaga aagctggaac tgaactttta gaaccatacc
     2821 tttcttttaa aatctatgca ccacaagaat acttaagtcg tgcttataac gatgcaccta
     2881 aatactgtgc taatattgtt gatactcaat taaagaacaa cgaagtaatt ttaagcggag
     2941 aaattcctgc acgttgtatt caagaatatc gtagtgattt aacatttttc actaatggac
     3001 gttctgtttg cttaactgaa ttaaaaggtt atcatgttac tactggtgaa cctgtatgcc
     3061 aaccacgtcg tcctaatagt cgtattgata aagttcgtta tatgttcaac aaaatcacat
     3121 aataatacta gtagcggccg ctgcagtaat acgactcact atagggcata cgatttaggt
     3181 gacactatag tagggataac agggtaattt agttacttac ctagcatgac attaacctat
     3241 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     3301 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3361 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     3421 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     3481 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     3541 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cagccaaacg tctcttcagg
     3601 ccactgacta gcgataactt tccccacaac ggaacaactc tcattgcatg ggatcattgg
     3661 gtactgtggg tttagtggtt gtaaaaacac ctgaccgcta tccctgatca gtttcttgaa
     3721 ggtaaactca tcacccccaa gtctggctat gcagaaatca cctggctcaa cagcctgctc
     3781 agggtcaacg agaattaaca ttccgtcagg aaagcttggc ttggagcctg ttggtgcggt
     3841 catggaatta ccttcaacct caagccagaa tgcagaatca ctggcttttt tggttgtgct
     3901 tacccatctc tccgcatcac ctttggtaaa ggttctaagc ttaggtgaga acatccctgc
     3961 ctgaacatga gaaaaaacag ggtactcata ctcacttcta agtgacggct gcatactaac
     4021 cgcttcatac atctcgtaga tttctctggc gattgaaggg ctaaattctt caacgctaac
     4081 tttgagaatt tttgtaagca atgcggcgtt ataagcattt aatgcattga tgccattaaa
     4141 taaagcacca acgcctgact gccccatccc catcttgtct gcgacagatt cctgggataa
     4201 gccaagttca tttttctttt tttcataaat tgctttaagg cgacgtgcgt cctcaagctg
     4261 ctcttgtgtt aatggtttct tttttgtgct cattagaaac tgtccttcgc atgattaaga
     4321 tgtttcagta cgaaaattgc tttcattgtt gatctccttt ttaagtgaac ttgggccctc
     4381 cacacattat acagaaaaat tttcctgatg tcaactggcg aaaagatctc tgcattaatg
     4441 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgc
//
