LOCUS       sequence-218182-        7787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK assembled broad-host-range plasmid encoding NanoLuc luciferase 
            protein driven by CP25 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7787)
  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 7787)
  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: pBTK563
FEATURES             Location/Qualifiers
     source          1..7787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             549..1064
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             complement(1222..2013)
                     /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"
     CDS             complement(2393..3244)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(3231..4070)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(4581..5552)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            6791..6878
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      7220..7614
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
ORIGIN
        1 tactggtcgt taccagagcc accgacccga gcaaaccctt ctctatcaga tcgttgacga
       61 gtattacccg gcattcgctg cgcttatggc agagcaggga aaggaattgc cgggctatgt
      121 gcaacgggaa tttgaagaat ttctccaatg cgggcggctg gagcatggct ttctacgggt
      181 tcgctgcgag tcttgccacg ccgagcacct ggtcgctttc agaaatcaat ctaaagtata
      241 tatgagtaaa cttggtctga caggcccctg aattcgcatc tagatggtag agccacaaac
      301 agccggtaca agcaacgatc tccaggacca tctgaatcat gcgcggatga cacgaactca
      361 cgacggcgat cacagacatt aacccacagt acagacactg cgacaacgtg gcaattcgtc
      421 gcaataccgt ctcactgaac tggccgataa ttgcagacga acgctttggc agtttattct
      481 tgacatgtag tgagggggct ggtataatca catagtactg ttatacagaa acagaggaga
      541 tattacatat ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct
      601 acaacctgga ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg
      661 tgtccgtaac tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca
      721 tccatgtcat catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa
      781 tttttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca
      841 cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag
      901 gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca
      961 aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca
     1021 acggagtgac cggctggcgg ctgtgcgaac gcattctggc gtaaatccgt aatcgttaat
     1081 ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag
     1141 agcatttgtc agctggatca gcagatggga accggatact cctcagcctc cagagtagcc
     1201 aatgagacgc cgagcggccg cttatttgcc gactaccttg gtgatctcgc ctttcacgta
     1261 gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag
     1321 ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc
     1381 ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg
     1441 ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag
     1501 cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc
     1561 ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc
     1621 cagatcaatg tcgatcgtgg ctggctcgaa gatacccgca agaatgtcat tgcgctgcca
     1681 ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac
     1741 aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc
     1801 caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac
     1861 cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg
     1921 tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg
     1981 agtcgatact tcggcgatca ccgcttccct cataatgttt aactttgttt tagggcgact
     2041 gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     2101 cgcttgctgc ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc
     2161 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     2221 gcgtgagcgc atacgctact tgcattacag cttacgaacc gaacaggctt atgtccactg
     2281 ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gtagcagcga
     2341 agtcgaggca tttctgtcct ggctggtcat gaccaaaatc ccttaacgtg agtcagcctg
     2401 ccgccttggg ccgggtgatg tcgtacttgc ccgccgcgaa ctcggttacc gtccagccca
     2461 gcgcgaccag ctccggcaac gcctcgcgca cccgctggcg gcgcttgcgc atggtcgaac
     2521 cactggcctc tgacggccag acatagccgc acaaggtatc tatggaagcc ttgccggttt
     2581 tgccggggtc gatccagcca cacagccgct ggtgcagcag gcgggcggtt tcgctgtcca
     2641 gcgcccgcac ctcgtccatg ctgatgcgca catgctggcc gccacccatg acggcctgcg
     2701 cgatcaaggg gttcagggcc acgtacaggc gcccgtccgc ctcgtcgctg gcgtactccg
     2761 acagcagccg aaacccctgc cgcttgcggc cattctgggc gatgatggat accttccaaa
     2821 ggcgctcgat gcagtcctgt atgtgcttga gcgccccacc actatcgacc tctgccccga
     2881 tttcctttgc cagcgcccga tagctacctt tgaccaccat ggcatcagcg gtgacggcct
     2941 cccacttggg ttccaggaac agccggagct gccgtccgcc ttcggtcttg ggttccgggc
     3001 caagcactag gccattaggc ccagccatgg ccaccagccc ttgcaggatg cgcagatcat
     3061 cagcgcccag cggctccggg ccgctgaact cgatccgctt gccgtcgccg tagtcatacg
     3121 tcacgtccag cttgctgcgc ttgcgctcgc cccgcttgag ggcacggaac aggccggggg
     3181 ccagacagtg cgccgggtcg tgccggacgt ggctgaggct gtgcttgttc ttaggcttca
     3241 ccacggggca cccccttgct cttgcgctgc ctctccagca cggcgggctt gagcaccccg
     3301 ccgtcatgcc gcctgaacca ccgatcagcg aacggtgcgc catagttggc cttgctcaca
     3361 ccgaagcgga cgaagaaccg gcgctggtcg tcgtccacac cccattcctc ggcctcggcg
     3421 ctggtcatgc tcgacaggta ggactgccag cggatgttat cgaccagtac cgagctgccc
     3481 cggctggcct gctgctggtc gcctgcgccc atcatggccg cgcccttgct ggcatggtgc
     3541 aggaacacga tagagcaccc ggtatcggcg gcgatggcct ccatgcgacc gatgacctgg
     3601 gccatggggc cgctggcgtt ttcttcctcg atgtggaacc ggcgcagcgt gtccagcacc
     3661 atcaggcggc ggccctcggc ggcgcgcttg aggccgtcga accactccgg ggccatgatg
     3721 ttgggcaggc tgccgatcag cggctggatc agcaggccgt cagccacggc ttgccgttcc
     3781 tcggcgctga ggtgcgcccc aagggcgtgc aggcggtgat gaatggcggt gggcgggtct
     3841 tcggcgggca ggtagatcac cgggccggtg ggcagttcgc ccacctccag cagatccggc
     3901 ccgcctgcaa tctgtgcggc cagttgcagg gccagcatgg atttaccggc accaccgggc
     3961 gacaccagcg ccccgaccgt accggccacc atgttgggca aaacgtagtc cagcggtggc
     4021 ggcgctgctg cgaacgcctc cagaatattg ataggcttat gggtagccat tgattgcctc
     4081 ctttgcaggc agttggtggt taggcgctgg cggggtcact acccccgccc tgcgccgctc
     4141 tgagttcttc caggcactcg cgcagcgcct cgtattcgtc gtcggtcagc cagaacttgc
     4201 gctgacgcat ccctttggcc ttcatgcgct cggcatatcg cgcttggcgt acagcgtcag
     4261 ggctggccag caggtcgccg gtctgcttgt ccttttggtc tttcatatca gtcaccgaga
     4321 aacttgccgg ggccgaaagg cttgtcttcg cggaacaagg acaaggtgca gccgtcaagg
     4381 ttaaggctgg ccatatcagc gactgaaaag cggccagcct cggccttgtt tgacgtataa
     4441 ccaaagccac cgggcaacca atagcccttg tcacttttga tcaggtagac cgaccctgaa
     4501 gcgctttttt cgtattccat aaaaccccct tctgtgcgtg agtactcata gtataacagg
     4561 cgtgagtacc aacgcaagca ctacatgctg aaatctggcc cgcccctgtc catgcctcgc
     4621 tggcggggtg ccggtgcccg tgccagctcg gcccgcgcaa gctggacgct gggcagaccc
     4681 atgaccttgc tgacggtgcg ctcgatgtaa tccgcttcgt ggccgggctt gcgctctgcc
     4741 agcgctgggc tggcctcggc catggccttg ccgatttcct cggcactgcg gccccggctg
     4801 gccagcttct gcgcggcgat aaagtcgcac ttgctgaggt catcaccgaa gcgcttgacc
     4861 agcccggcca tctcgctgcg gtactcgtcc agcgccgtgc gccggtggcg gctaagctgc
     4921 cgctcgggca gttcgaggct ggccagcctg cgggccttct cctgctgccg ctgggcctgc
     4981 tcgatctgct ggccagcctg ctgcaccagc gccgggccag cggtggcggt cttgcccttg
     5041 gattcacgca gcagcaccca cggctgataa ccggcgcggg tggtgtgctt gtccttgcgg
     5101 ttggtgaagc ccgccaagcg gccatagtgg cggctgtcgg cgctggccgg gtcggcgtcg
     5161 tactcgctgg ccagcgtccg ggcaatctgc ccccgaagtt caccgcctgc ggcgtcggcc
     5221 accttgaccc atgcctgata gttcttcggg ctggtttcca ctaccagggc aggctcccgg
     5281 ccctcggctt tcatgtcatc caggtcaaac tcgctgaggt cgtccaccag caccagacca
     5341 tgccgctcct gctcggcggg cctgatatac acgtcattgc cctgggcatt catccgcttg
     5401 agccatggcg tgttctggag cacttcggcg gctgaccatt cccggttcat catctggccg
     5461 gtggtggcgt ccctgacgcc gatatcgaag cgctcacagc ccatggcctt gagctgtcgg
     5521 cctatggcct gcaaagtcct gtcgttcttc atcgggccac caagcgcagc cagatcgagc
     5581 cgtcctcggt tgtcagtggc gtcaggtcga gcaagagcaa cgatgcgatc agcagcacca
     5641 ccgtaggcat catggaagcc agcatcacgg ttagccatag cttccagtgc cacccccgcg
     5701 acgcgctccg ggcgctctgc gcggcgctgc tcacctcggc ggctacctcc cgcaactctt
     5761 tggccagctc cacccatgcc gcccctgtct ggcgctgggc tttcagccac tccgccgcct
     5821 gcgcctcgct ggcctgctgg gtctggctca tgacctgccg ggcttcgtcg gccagtgtcg
     5881 ccatgctctg ggccagcggt tcgatctgct ccgctaactc gttgatgcct ctggatttct
     5941 tcactctgtc gattgcgttc atggtctatt gcctcccggt attcctgtaa gtcgatgatc
     6001 tgggcgttgg cggtgtcgat gttcagggcc acgtctgccc ggtcggtgcg gatgccccgg
     6061 ccttccatct ccaccacgtt cggccccagg tgaacaccgg gcaggcgctc gatgccctgc
     6121 gcctcaagtg ttctgtggtc aatgcgggcg tcgtggccag cccgctctaa tgcccggttg
     6181 gcatggtcgg cccatgcctc gcgggtctgc tcaagccatg ccttgggctt gagcgcttcg
     6241 gtcttctgtg ccccgccctt ctccggggtc ttgccgttgt accgcttgaa ccactgagcg
     6301 gcgggccgct cgatgccgtc attgatccgc tcggagatca tcaggtggca gtgcgggttc
     6361 tcgccgccac cggcatggat ggccagcgta tacggcaggc gctcggcacc ggtcaggtgc
     6421 tgggcgaact cggacgccag cgccttctgc tggtcgaggg tcagctcgac cggcagggca
     6481 aattcgacct ccttgaacag ccgcccattg gcgcgttcat acaggtcggc agcatcccag
     6541 tagtcggcgg gccgctcgac gaactccggc atgtgcccgg attcggcgtg caagacttca
     6601 tccatgtcgc gggcatactt gccttcgcgc tggatgtagt cggccttggc cctggccgat
     6661 tggccgcccg acctgctgcc ggttttcgcc gtaaggtgat aaatcgccat gctgcctcgc
     6721 tgttgctttt gcttttcggc tccatgcaat ggccctcgga gagcgcaccg cccgaagggt
     6781 ggccgttagg ccagtttctc gaagagaaac cggtaagtgc gccctcccct acaaagtagg
     6841 gtcgggattg ccgccgctgt gcctccatga tagcctacga gacagcacat taacaatggg
     6901 gtgtcaagat ggttaagggg agcaacaagg cggcggatcg gctggccaag ctcgaagaac
     6961 aacgagcgcg aatcaatgcc gaaattcagc gggtgcgggc aagggaacag cagcaagagc
     7021 gcaagaacga aacaaggcgc aaggtgctgg tgggggccat gattttggcc aaggtgaaca
     7081 gcagcgagtg gccggaggat cggctcatgg cggcaatgga tgcgtacctt gaacgcgacc
     7141 acgaccgcgc cttgttcggt ctgccgccac gccagaagga tgagccgggc tgaatgatcg
     7201 accgagacag gccctgcggg gctgcacacg cgcccccacc cttcgggtag ggggaaaggc
     7261 cgctaaagcg gctaaaagcg ctccagcgta tttctgcggg gtttggtgtg gggtttagcg
     7321 ggctttgccc gcctttcccc ctgccgcgca gcggtggggc ggtgtgtagc ctagcgcagc
     7381 gaatagacca gctatccggc ctctggccgg gcatattggg caagggcagc agcgccccac
     7441 aagggcgctg ataaccgcgc ctagtggatt attcttagat aatcatggat ggatttttcc
     7501 aacaccccgc cagcccccgc ccctgctggg tttgcaggtt tgggggcgtg acagttattg
     7561 caggggttcg tgacagttat tgcagggggg cgtgacagtt attgcagggg ttcgtgacag
     7621 ttagtacggg agtgacgggc actggctggc aatgtctagc aacggcaggc atttcggctg
     7681 agggtaaaag aactttccgc taagcgatag actgtatgta aacacagtat tgcaaggacg
     7741 cggaacatgc ctcatgtggc ggccaggacg gccagccggg atcggga
//
