LOCUS       sequence-208073-        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             complement(27..878)
                     /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
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRKRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(865..1704)
                     /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(2215..3186)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDAPTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGHDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            4425..4512
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      4854..5248
                     /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)"
     CDS             5970..6485
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             complement(6643..7434)
                     /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 tcatgaccaa aatcccttaa cgtgagtcag cctgccgcct tgggccgggt gatgtcgtac
       61 ttgcccgccg cgaactcggt taccgtccag cccagcgcga ccagctccgg caacgcctcg
      121 cgcacccgct tgcggcgctt gcgcatggtc gaaccactgg cctctgacgg ccagacatag
      181 ccgcacaagg tatctatgga agccttgccg gttttgccgg ggtcgatcca gccacacagc
      241 cgctggtgca gcaggcgggc ggtttcgctg tccagcgccc gcacctcgtc catgctgatg
      301 cgcacatgct ggccgccacc catgacggcc tgcgcgatca aggggttcag ggccacgtac
      361 aggcgcccgt ccgcctcgtc gctggcgtac tccgacagca gccgaaaccc ctgccgcttg
      421 cggccattct gggcgatgat ggataccttc caaaggcgct cgatgcagtc ctgtatgtgc
      481 ttgagcgccc caccactatc gacctctgcc ccgatttcct ttgccagcgc ccgatagcta
      541 cctttgacca catggcattc agcggtgacg gcctcccact tgggttccag gaacagccgg
      601 agctgccgtc cgccttcggt cttgggttcc gggccaagca ctaggccatt aggcccagcc
      661 atggccacca gcccttgcag gatgcgcaga tcatcagcgc ccagcggctc cgggccgctg
      721 aactcgatcc gcttgccgtc gccgtagtca tacgtcacgt ccagcttgct gcgcttgcgc
      781 tcgccccgct tgagggcacg gaacaggccg ggggccagac agtgcgccgg gtcgtgccgg
      841 acgtggctga ggctgtgctt gttcttaggc ttcaccacgg ggcaccccct tgctcttgcg
      901 ctgcctctcc agcacggcgg gcttgagcac cccgccgtca tgccgcctga accaccgatc
      961 agcgaacggt gcgccatagt tggccttgct cacaccgaag cggacgaaga accggcgctg
     1021 gtcgtcgtcc acaccccatt cctcggcctc ggcgctggtc atgctcgaca ggtaggactg
     1081 ccagcggatg ttatcgacca gtaccgagct gccccggctg gcctgctgct ggtcgcctgc
     1141 gcccatcatg gccgcgccct tgctggcatg gtgcaggaac acgatagagc acccggtatc
     1201 ggcggcgatg gcctccatgc gaccgatgac ctgggccatg gggccgctgg cgttttcttc
     1261 ctcgatgtgg aaccggcgca gcgtgtccag caccatcagg cggcggccct cggcggcgcg
     1321 cttgaggccg tcgaaccact ccggggccat gatgttgggc aggctgccga tcagcggctg
     1381 gatcagcagg ccgtcagcca cggcttgccg ttcctcggcg ctgaggtgcg ccccaagggc
     1441 gtgcaggcgg tgatgaatgg cggtgggcgg gtcttcggcg ggcaggtaga tcaccgggcc
     1501 ggtgggcagt tcgcccacct ccagcagatc cggcccgcct gcaatctgtg cggccagttg
     1561 cagggccagc atggatttac cggcaccacc gggcgacacc agcgccccga ccgtaccggc
     1621 caccatgttg ggcaaaacgt agtccagcgg tggcggcgct gctgcgaacg cctccagaat
     1681 attgataggc ttatgggtag ccattgattg cctcctttgc aggcagttgg tggttaggcg
     1741 ctggcggggt cactaccccc gccctgcgcc gctctgagtt cttccaggca ctcgcgcagc
     1801 gcctcgtatt cgtcgtcggt cagccagaac ttgcgctgac gcatcccttt ggccttcatg
     1861 cgctcggcat atcgcgcttg gcgtacagcg tcagggctgg ccagcaggtc gccggtctgc
     1921 ttgtcctttt ggtctttcat atcagtcacc gagaaacttg ccggggccga aaggcttgtc
     1981 ttcgcggaac aaggacaagg tgcagccgtc aaggttaagg ctggccatat cagcgactga
     2041 aaagcggcca gcctcggcct tgtttgacgt ataaccaaag ccaccgggca accaatagcc
     2101 cttgtcactt ttgatcaggt agaccgaccc tgaagcgctt ttttcgtatt ccataaaacc
     2161 cccttctgtg cgtgagtact catagtataa caggcgtgag taccaacgca agcactacat
     2221 gctgaaatct ggcccgcccc tgtccatgcc tcgctggcgg ggtgccggtg cccgtgccag
     2281 ctcggcccgc gcaagctgga cgctgggcag acccatgacc ttgctgacgg tgcgctcgat
     2341 gtaatccgct tcgtggccgg gcttgcgctc tgccagcgct gggctggcct cggccatggc
     2401 cttgccgatt tcctcggcac tgcggccccg gctggccagc ttctgcgcgg cgataaagtc
     2461 gcacttgctg aggtcatgac cgaagcgctt gaccagcccg gccatctcgc tgcggtactc
     2521 gtccagcgcc gtgcgccggt ggcggctaag ctgccgctcg ggcagttcga ggctggccag
     2581 cctgcgggcc ttctcctgct gccgctgggc ctgctcgatc tgctggccag cctgctgcac
     2641 cagcgccggg ccagcggtgg cggtcttgcc cttggattca cgcagcagca cccacggctg
     2701 ataaccggcg cgggtggtgt gcttgtcctt gcggttggtg aagcccgcca agcggccata
     2761 gtggcggctg tcggcgctgg ccgggtcggc gtcgtactcg ctggccagcg tccgggcaat
     2821 ctgcccccga agttcaccgc ctgcggcgtc ggccaccttg acccatgcct gatagttctt
     2881 cgggctggtt tccactacca gggcaggctc ccggccctcg gctttcatgt catccaggtc
     2941 aaactcgctg aggtcgtcca ccagcaccag accatgccgc tcctgctcgg cgggcctgat
     3001 atacacgtca ttgccctggg cattcatccg cttgagccat ggcgtgttct ggagcacttc
     3061 ggcggctgac cattcccggt tcatcatctg gccggtgggt gcgtccctga cgccgatatc
     3121 gaagcgctca cagcccatgg ccttgagctg tcggcctatg gcctgcaaag tcctgtcgtt
     3181 cttcatcggg ccaccaagcg cagccagatc gagccgtcct cggttgtcag tggcgtcagg
     3241 tcgagcaaga gcaacgatgc gatcagcagc accaccgtag gcatcatgga agccagcatc
     3301 acggttagcc atagcttcca gtgccacccc cgcgacgcgc tccgggcgct ctgcgcggcg
     3361 ctgctcacct cggcggctac ctcccgcaac tctttggcca gctccaccca tgccgcccct
     3421 gtctggcgct gggctttcag ccactccgcc gcctgcgcct cgctggcctg cttggtctgg
     3481 ctcatgacct gccgggcttc gtcggccagt gtcgccatgc tctgggccag cggttcgatc
     3541 tgctccgcta actcgttgat gcctctggat ttcttcactc tgtcgattgc gttcatggtc
     3601 tattgcctcc cggtattcct gtaagtcgat gatctgggcg ttggcggtgt cgatgttcag
     3661 ggccacgtct gcccggtcgg tgcggatgcc ccggccttcc atctccacca cgttcggccc
     3721 caggtgaaca ccgggcaggc gctcgatgcc ctgcgcctca agtgttctgt ggtcaatgcg
     3781 ggcgtcgtgg ccagcccgct ctaatgcccg gttggcatgg tcggcccatg cctcgcgggt
     3841 ctgctcaagc catgccttgg gcttgagcgc ttcggtcttc tgtgccccgc ccttctccgg
     3901 ggtcttgccg ttgtaccgct tgaaccactg agcggcgggc cgctcgatgc cgtcattgat
     3961 ccgctcggag atcatcaggt ggcagtgcgg gttctcgccg ccaccggcat ggatggccag
     4021 cgtatacggc aggcgctcgg caccggtcag gtgctgggcg aactcggacg ccagcgcctt
     4081 ctgctggtcg agggtcagct cgaccggcag ggcaaattcg acctccttga acagccgccc
     4141 attggcgcgt tcatacaggt cggcagcatc ccagtagtcg gcgggccgct cgacgaactc
     4201 cggcatgtgc ccggattcgg cgtgcaagac ttcatccatg tcgcgggcat acttgccttc
     4261 gcgctggatg tagtcggcct tggccctggc cgattggccg cccgacctgc tgccggtttt
     4321 cgccgtaagg tgataaatcg ccatgctgcc tcgctgttgc ttttgctttt cggctccatg
     4381 caatggccct cggagagcgc accgcccgaa gggtggccgt taggccagtt tctcgaagag
     4441 aaaccggtaa gtgcgccctc ccctacaaag tagggtcggg attgccgccg ctgtgcctcc
     4501 atgatagcct acgagacagc acattaacaa tggggtgtca agatggttaa ggggagcaac
     4561 aaggcggcgg atcggctggc caagctcgaa gaacaacgag cgcgaatcaa tgccgaaatt
     4621 cagcgggagc gggcaaggga acagcagcaa gagcgcaaga acgaaacaag gcgcaaggtg
     4681 ctggtggggg ccatgatttt ggccaaggtg aacagcagcg agtggccgga ggatcggctc
     4741 atggcggcaa tggatgcgta ccttgaacgc gaccacgacc gcgccttgtt cggtctgccg
     4801 ccacgccaga aggatgagcc gggctgaatg atcgaccgag acaggccctg cggggctgca
     4861 cacgcgcccc cacccttcgg gtagggggaa aggccgctaa agcggctaaa agcgctccag
     4921 cgtatttctg cggggtttgg tgtggggttt agcgggcttt gcccgccttt ccccctgccg
     4981 cgcagcggtg gggcggtgtg tagcctagcg cagcgaatag accagctatc cggcctctgg
     5041 ccgggcatat tgggcaaggg cagcagcgcc ccacaagggc gctgataacc gcgcctagtg
     5101 gattattctt agataatcat ggatggattt ttccaacacc ccgccagccc ccgcccctgc
     5161 tgggtttgca ggtttggggg cgtgacagtt attgcagggg ttcgtgacag ttattgcagg
     5221 ggggcgtgac agttattgca ggggttcgtg acagttagta cgggagtgac gggcactggc
     5281 tggcaatgtc tagcaacggc aggcatttcg gctgagggta aaagaacttt ccgctaagcg
     5341 atagactgta tgtaaacaca gtattgcaag gacgcggaac atgcctcatg tggcggccag
     5401 gacggccagc cgggatcggg atactggtcg ttaccagagc caccgacccg agcaaaccct
     5461 tctctatcag atcgttgacg agtattaccc ggcattcgct gcgcttatgg cagagcaggg
     5521 aaaggaattg ccgggctatg tgcaacggga atttgaagaa tttctccaat gcgggcggct
     5581 ggagcatggc tttctacggg ttcgctgcga gtcttgccac gccgagcacc tggtcgcttt
     5641 cagaaatcaa tctaaagtat atatgagtaa acttggtctg acaggcccct gaattcgcat
     5701 ctagatggta gagccacaaa cagccggtac aagcaacgat ctccaggacc atctgaatca
     5761 tgcgcggatg acacgaactc acgacggcga tcacagacat taacccacag tacagacact
     5821 gcgacaacgt ggcaattcgt cgcaataccg tctcactgaa ctggccgata attgcagacg
     5881 aacgctttgg cagtttattc ttgacatgta gtgagggggc tggtataatc acatagtact
     5941 gttatacaga aacagaggag atattacata tggtcttcac actcgaagat ttcgttgggg
     6001 actggcgaca gacagccggc tacaacctgg accaagtcct tgaacaggga ggtgtgtcca
     6061 gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg
     6121 aaaatgggct gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc
     6181 aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta
     6241 aggtgatcct gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact
     6301 atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag
     6361 ggaccctgtg gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc
     6421 tgctgttccg agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg
     6481 cgtaaatccg taatcgttaa tccgcaaata acgtaaaaac ccgcttcggc gggttttttt
     6541 atggggggag tttagggaaa gagcatttgt cagctggatc agcagatggg aaccggatac
     6601 tcctcagcct ccagagtagc caatgagacg ccgagcggcc gcttatttgc cgactacctt
     6661 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
     6721 gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
     6781 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     6841 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     6901 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     6961 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     7021 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacccgc
     7081 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     7141 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     7201 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     7261 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc
     7321 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac
     7381 gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcataatgtt
     7441 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa
     7501 acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacccc
     7561 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc
     7621 ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca gcttacgaac
     7681 cgaacaggct tatgtccact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc
     7741 ggcaaccttg ggtagcagcg aagtcgaggc atttctgtcc tggctgg
//
