LOCUS       sequence-254616-        3383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 8 Bacterial origin (R6K suicide plasmid) for homologous 
            recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3383)
  AUTHORS   Leonard SP, Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar 
            S, Davies BW, Ellington AD, Moran NA, Barrick JE
  TITLE     Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit 
            for Modular Assembly of Broad-Host-Range Plasmids.
  JOURNAL   ACS Synth Biol. 2018 May 18;7(5):1279-1290. doi: 
            10.1021/acssynbio.7b00399. Epub 2018 Apr 13.
  PUBMED    29608282
REFERENCE   2  (bases 1 to 3383)
  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"; doi: 
            "10.1021/acssynbio.7b00399"; journalName: "ACS Synth Biol"; date: 
            "2018-05-18- 18"; volume: "7"; issue: "5"; pages: "1279-1290"
COMMENT     Sequence Label: pBTK599s
FEATURES             Location/Qualifiers
     source          1..3383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             795..806
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             813..1526
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1538..1609
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1625..1652
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     oriT            1768..1877
                     /note="incP origin of transfer"
     rep_origin      2048..2436
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     2469..2485
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2469..2485
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2519..2535)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2520..2536)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(join(2854..3383,1..262))
                     /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"
ORIGIN
        1 cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt
       61 tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa
      121 tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca
      181 acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt
      241 cggcgatcac cgcttccctc ataatgttta actttgtttt agggcgactg ccctgctgcg
      301 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
      361 tggatgcccg aggcatagac tgtaccccaa aaaaacagtc ataacaagcc atgaaaaccg
      421 ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca
      481 tacgctactt gcattacagc ttacgaaccg aacaggctta tgtccactgg gttcgtgcct
      541 tcatccgttt ccacggtgtg cgtcacccgg caaccttggg tagcagcgaa gtcgtggcat
      601 ttctgtcctg gctggccgaa aagtgccacc tgcctgatga gacgatgaaa gtgaaacgtg
      661 atttcatgcg tcattttgaa cattttgtaa atcttattta ataatgtgtg cggcaattca
      721 catttaattt atgaatgttt tcttaacatc gcggcaactc aagaaacggc aggttcggat
      781 cttagctact agagaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg
      841 gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc
      901 gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta
      961 ctggtaaact gccggttcct tggccgactc tggtaacgac gctgacttat ggtgttcagt
     1021 gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg
     1081 aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg
     1141 cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact
     1201 ttaaagagga cggcaatatc ctgggccata agctggaata caattttaac agccacaatg
     1261 tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca
     1321 acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg
     1381 atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag
     1441 atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca
     1501 cgcatggtat ggatgaactg tacaaatgac caggcatcaa ataaaacgaa aggctcagtc
     1561 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     1621 cactggctca ccttcgggtg ggcctttctg cgtttataag tccgtctcaa gcaaggggga
     1681 tcaattccgt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct
     1741 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga
     1801 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
     1861 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
     1921 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
     1981 tgaccccgaa gcagggttat gcagcggaaa acggaattga tccggccacg atgcgtccgg
     2041 cgtagaggat ctgaagatca gcagttcaac ctgttgatag tacgtactaa gctctcatgt
     2101 ttcacgtact aagctctcat gtttaacgta ctaagctctc atgtttaacg aactaaaccc
     2161 tcatggctaa cgtactaagc tctcatggct aacgtactaa gctctcatgt ttcacgtact
     2221 aagctctcat gtttgaacaa taaaattaat ataaatcagc aacttaaata gcctctaagg
     2281 ttttaagttt tataagaaaa aaaagaatat ataaggcttt taaagctttt aaggtttaac
     2341 ggttgtggac aacaagccag ggatgtaacg cactgagaag cccttagagc ctctcaaagc
     2401 aattttgagt gacacaggaa cacttaacgg ctgacatggg aattcccctc caccgcggtg
     2461 gcggccgctc tagaactagt ggatcccccg ggctgcagga attcgatatc aagcttatcg
     2521 ataccgtcga cctcgagggg gggcccggta ccgaggacgc gtcgaataat tccgctagct
     2581 tcacgctgcc gcaagcactc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa
     2641 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca
     2701 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggattat caaaaaggat
     2761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2821 gtaaacttgg tctgacagtt accgagcggc cgcttatttg ccgactacct tggtgatctc
     2881 gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc
     2941 ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag
     3001 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
     3061 gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg
     3121 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
     3181 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
     3241 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacccg caagaatgtc
     3301 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat
     3361 gtcgtcgtgc acaacaatgg tga
//
