LOCUS       sequence-416212-        5653 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Suicide plasmid carrying a neomycin resistance marker flanked by 
            homologous regions of the Mth60-fimbria encoding operon of 
            Methanothermobacter thermautotrophicus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5653)
  AUTHORS   Fink C, Martinez-Cano G, Shuster J, Panzera A, Rennhack KE, Rohbohm 
            N, Angenent LT, Molitor B
  TITLE     The Targeted Deletion of Genes Responsible for Expression of the 
            Mth60 Fimbriae Leads to Loss of Cell-Cell Connections in 
            Methanothermobacter thermautotrophicus DeltaH.
  JOURNAL   Appl Environ Microbiol. 2023 Jul 26;89(7):e0057523. doi: 
            10.1128/aem.00575-23. Epub 2023 Jun 13.
  PUBMED    37310347
REFERENCE   2  (bases 1 to 5653)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol."; date: "2023-07-26"; volume: "89(7):e0057523. 
            doi"; pages: " 10.1128/aem.00575-23. Epub 2023 Jun 13"
COMMENT     Sequence Label: pCF702
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1711..2299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2200..2219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2453..2470
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            2620..2729
                     /note="incP origin of transfer"
     CDS             2762..3133
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     primer_bind     3330..3346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3330..3346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4494..5288
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 cgcgccgaaa tcccaacctt catataatat tgcaagaaca cctttttata taattttcga
       61 taggtcggga tgtcttcctt attcctggac tagagggtgc ggatgatcat cctcctcttc
      121 gtgtgtctcc tgaagtcgag ggatgttgat accctgctga ggtacctcct gctgatccta
      181 gctgcctctc tgatttcatc gtcacggggt ctgcgggcgt cgaggctgtt accatgggca
      241 acataccctg ttatcctgta gggaacatcc catactgaca ggaactctgc aatcccctca
      301 atctcatcaa ggtcaacaaa gccaggcacg aggaccgttg agacctcaat cctctctgaa
      361 cctgagaact cctctatgca ttcaaggacc tccctgttgg actccccggt cagctggatg
      421 tgtttctcct cgtcgagggc cttgaggtca acgtggacct catggaaggt gctcctcctt
      481 agaacatccc tgcgggtgcc gttggttgat attgtcacat ggaagtctcc tgcgaggatt
      541 tcggtcagtt caggaaggtc ctcctggagt gtgggttcac cccctgctat gaggaccctc
      601 tctaccccgg tttcatcacg gatcctctgg attagatctg ctatcctgtc tggggagtgg
      661 ttcatacatc ctgatggttt gaaaaagcag tacctgcacc tgaagttgca tgttggtgtc
      721 aggacggtta tgatgtcgct tacagatatg tgtgagagct ggatgtgttt cacctctgga
      781 tggcatgttt ttatatctgt ggtatgtgtt atggcccggt cgacagtggg caagttgaaa
      841 aattcacaaa aatgtggtat aatatctttg ttcattagag cgataaactt gaatttgaga
      901 gggaacttag atggtatttg aaaaaattga taaaaatagt tggaacagaa aagagtattt
      961 tgaccactac tttgcaagtg taccttgtac ctacagcatg accgttaaag tggatatcac
     1021 acaaataaag gaaaagggaa tgaaactata tcctgcaatg ctttattata ttgcaatgat
     1081 tgtaaaccgc cattcagagt ttaggacggc aatcaatcaa gatggtgaat tggggatata
     1141 tgatgagatg ataccaagct atacaatatt tcacaatgat actgaaacat tttccagcct
     1201 ttggactgag tgtaagtctg actttaaatc atttttagca gattatgaaa gtgatacgca
     1261 acggtatgga aacaatcata gaatggaagg aaagccaaat gctccggaaa acatttttaa
     1321 tgtatctatg ataccgtggt caaccttcga tggctttaat ctgaatttgc agaaaggata
     1381 tgattatttg attcctattt ttactatggg gaaatattat aaagaagata acaaaattat
     1441 acttcctttg gcaattcaag ttcatcacgc agtatgtgac ggatttcaca tttgccgttt
     1501 tgtaaacgaa ttgcaggaat tgataaatag ttaacttcag gtttgtctgt aactaaaaac
     1561 aagtatttaa gcaaaaacat cgtagaaata cggtgttttt tgttacccta agtttaaact
     1621 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1681 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1741 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1801 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1861 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1921 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1981 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2041 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2101 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2161 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2221 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2281 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2341 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2401 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2461 agtgagcgag gaagcggaag agcgcccaat acgcagggcc ccctgcttcg gggtcattat
     2521 agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt
     2581 tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg
     2641 cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa
     2701 cgggaatcct gctctgcgag gctggccggc taccgccggc gtaacagatg agggcaagcg
     2761 gatggctgat gaaaccaagc caaccaggaa gggcagccca cctatcaagg tgtactgcct
     2821 tccagacgaa cgaagagcga ttgaggaaaa ggcggcggcg gccggcatga gcctgtcggc
     2881 ctacctgctg gccgtcggcc agggctacaa aatcacgggc gtcgtggact atgagcacgt
     2941 ccgcgagctg gcccgcatca atggcgacct gggccgcctg ggcggcctgc tgaaactctg
     3001 gctcaccgac gacccgcgca cggcgcggtt cggtgatgcc acgatcctcg ccctgctggc
     3061 gaagatcgaa gagaagcagg acgagcttgg caaggtcatg atgggcgtgg tccgcccgag
     3121 ggcagagcca tgactttttt agccgctaaa acggccgggg ggtgcgcgtg attgccaagc
     3181 acgtccccat gcgctccatc aagaagagcg acttcgcgga gctggtgaag tacatcaccg
     3241 acgagcaagg caagaccgat cgggccccct gcaggataaa aaaattgtag ataaatttta
     3301 taaaatagtt ttatctacaa tttttttatc aggaaacagc tatgaccgcg gccgctgtat
     3361 ccatatgggt actgtgtgag ggtcatattc tgtgagaccg ttccaatgta gatgggtgtc
     3421 atgctggcgt ttttgaagta ggcgagtatg aggtcgttac ctttccttac tatgaatgcg
     3481 aagtccagtg gatcgagtct ggtttttcgg agcatcagga ggtttccttt accatagctc
     3541 actataccgc gggcctggtt gagtataccc tccagggcgt cctctgtcgt ctggtttttg
     3601 tagtatgctg ttcctgctgt tgttatgacc aggacctcac cggttctggt gaagttgagg
     3661 atctggtctg ccttctttgt tgtttccctt cccagtttgt aggcttctgt ggcgttcatg
     3721 gtgagtgaaa gatgggctgc gtataccgtg ccgtagtctg ttagctggaa ggtatgtgag
     3781 agttccctgt atccgggggc acggattttt actgtgaatt ttgtggtgtt tgtgatgttc
     3841 tggtagttga atttgatttt ggtcatgttt gctgccgggt caaatgttct ggtgaagttt
     3901 atctttgtcc cgttttcgtc tgtcagttcc tggatgtcag ggttgatatt gccattatcg
     3961 tcggcgtatt catatttgac gtcaaccccc acctcacagt ttgtatcagc tgcactcacc
     4021 gaacctgtaa gtgccacagc aacgagaaga accatgaaaa ccatgaaata ctgtctttcc
     4081 ataccttttt cacctccttt taattttatc attaaaaata atgatcttac taacatataa
     4141 gtcttaatta ttactacttg ggtgatgatg ggatgtattg taatactcac tgctttaaaa
     4201 ggagaactgt atggaattat cctgagatgc tcagaacaaa atagttttag aagaactgca
     4261 taaagtgatc aattaactga gaatatggta gaaaagaaaa aaggatggat tactttttcc
     4321 tctcgatctt gatcgcgatt ggaggttcaa ttcccttctt acggaggccg gccgcccatt
     4381 ttttgagtat tccccatgaa ccaaccgatg gctcagaaaa accttaaaat tagcgatata
     4441 tttatatagg attatatgaa tagataatat cacataaaat gaggtggtta attatggcta
     4501 aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc gtaaaagata
     4561 cggaaggaat gtctcctgct aaggtatata agctggtggg agaaaatgaa aacctatatt
     4621 taaaaatgac ggacagccgg tataaaggga ccacctatga tgtggaacgg gaaaaggaca
     4681 tgatgctatg gctggaagga aagctgcctg ttccaaaggt cctgcacttt gaacggcatg
     4741 atggctggag caatctgctc atgagtgagg ccgatggcgt cctttgctcg gaagagtatg
     4801 aagatgaaca aagccctgaa aagattatcg agctgtatgc ggagtgcatc aggctctttc
     4861 actccatcga catatcggat tgtccctata cgaatagctt agacagccgc ttagccgaat
     4921 tggattactt actgaataac gatctggccg atgtggattg cgaaaactgg gaagaagaca
     4981 ctccatttaa agatccgcgc gagctgtatg attttttaaa gacggaaaag cccgaagagg
     5041 aacttgtctt ttcccacggc gacctgggag acagcaacat ctttgtgaaa gatggcaaag
     5101 taagtggctt tattgatctt gggagaagcg gcagggcgga caagtggtat gacattgcct
     5161 tctgcgtccg gtcgatcagg gaggatatcg gggaagaaca gtatgtcgag ctattttttg
     5221 acttactggg gatcaagcct gattgggaga aaataaaata ttatatttta ctggatgaat
     5281 tgttttagcg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
     5341 gctccgaccg aagccacccg gggcggcccc gccgaccccg cacccgcccc cgaggcccac
     5401 cgcgggggac acaccgaaca cgccgaccct gctgaacacg cggcgcagtt cggtgcccag
     5461 gagcggatcg ggaattaatt cgaagctgct ggtgaaagag accctatctt acctgctaaa
     5521 atctaagtta attactaatt tattattaat ttattattag attgggcaaa atagtaaaag
     5581 aaaactaaag gaaacctaat atggtttcct ttttttatat atttttaatt cactgggggc
     5641 aattctgtca ggg
//
