LOCUS       sequence-218137-        7798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK assembled broad-host-range plasmid encoding NanoLuc luciferase 
            protein driven by PA1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7798)
  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 7798)
  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: pBTK564
FEATURES             Location/Qualifiers
     source          1..7798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(368..1219)
                     /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(1206..2045)
                     /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(2556..3527)
                     /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            4766..4853
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      5195..5589
                     /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             6322..6837
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             complement(6995..7786)
                     /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 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg
       61 acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta ccccaaaaaa
      121 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
      181 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagcttac gaaccgaaca
      241 ggcttatgtc cactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
      301 cttgggtagc agcgaagtcg aggcatttct gtcctggctg gtcatgacca aaatccctta
      361 acgtgagtca gcctgccgcc ttgggccggg tgatgtcgta cttgcccgcc gcgaactcgg
      421 ttaccgtcca gcccagcgcg accagctccg gcaacgcctc gcgcacccgc tggcggcgct
      481 tgcgcatggt cgaaccactg gcctctgacg gccagacata gccgcacaag gtatctatgg
      541 aagccttgcc ggttttgccg gggtcgatcc agccacacag ccgctggtgc agcaggcggg
      601 cggtttcgct gtccagcgcc cgcacctcgt ccatgctgat gcgcacatgc tggccgccac
      661 ccatgacggc ctgcgcgatc aaggggttca gggccacgta caggcgcccg tccgcctcgt
      721 cgctggcgta ctccgacagc agccgaaacc cctgccgctt gcggccattc tgggcgatga
      781 tggatacctt ccaaaggcgc tcgatgcagt cctgtatgtg cttgagcgcc ccaccactat
      841 cgacctctgc cccgatttcc tttgccagcg cccgatagct acctttgacc accatggcat
      901 cagcggtgac ggcctcccac ttgggttcca ggaacagccg gagctgccgt ccgccttcgg
      961 tcttgggttc cgggccaagc actaggccat taggcccagc catggccacc agcccttgca
     1021 ggatgcgcag atcatcagcg cccagcggct ccgggccgct gaactcgatc cgcttgccgt
     1081 cgccgtagtc atacgtcacg tccagcttgc tgcgcttgcg ctcgccccgc ttgagggcac
     1141 ggaacaggcc gggggccaga cagtgcgccg ggtcgtgccg gacgtggctg aggctgtgct
     1201 tgttcttagg cttcaccacg gggcaccccc ttgctcttgc gctgcctctc cagcacggcg
     1261 ggcttgagca ccccgccgtc atgccgcctg aaccaccgat cagcgaacgg tgcgccatag
     1321 ttggccttgc tcacaccgaa gcggacgaag aaccggcgct ggtcgtcgtc cacaccccat
     1381 tcctcggcct cggcgctggt catgctcgac aggtaggact gccagcggat gttatcgacc
     1441 agtaccgagc tgccccggct ggcctgctgc tggtcgcctg cgcccatcat ggccgcgccc
     1501 ttgctggcat ggtgcaggaa cacgatagag cacccggtat cggcggcgat ggcctccatg
     1561 cgaccgatga cctgggccat ggggccgctg gcgttttctt cctcgatgtg gaaccggcgc
     1621 agcgtgtcca gcaccatcag gcggcggccc tcggcggcgc gcttgaggcc gtcgaaccac
     1681 tccggggcca tgatgttggg caggctgccg atcagcggct ggatcagcag gccgtcagcc
     1741 acggcttgcc gttcctcggc gctgaggtgc gccccaaggg cgtgcaggcg gtgatgaatg
     1801 gcggtgggcg ggtcttcggc gggcaggtag atcaccgggc cggtgggcag ttcgcccacc
     1861 tccagcagat ccggcccgcc tgcaatctgt gcggccagtt gcagggccag catggattta
     1921 ccggcaccac cgggcgacac cagcgccccg accgtaccgg ccaccatgtt gggcaaaacg
     1981 tagtccagcg gtggcggcgc tgctgcgaac gcctccagaa tattgatagg cttatgggta
     2041 gccattgatt gcctcctttg caggcagttg gtggttaggc gctggcgggg tcactacccc
     2101 cgccctgcgc cgctctgagt tcttccaggc actcgcgcag cgcctcgtat tcgtcgtcgg
     2161 tcagccagaa cttgcgctga cgcatccctt tggccttcat gcgctcggca tatcgcgctt
     2221 ggcgtacagc gtcagggctg gccagcaggt cgccggtctg cttgtccttt tggtctttca
     2281 tatcagtcac cgagaaactt gccggggccg aaaggcttgt cttcgcggaa caaggacaag
     2341 gtgcagccgt caaggttaag gctggccata tcagcgactg aaaagcggcc agcctcggcc
     2401 ttgtttgacg tataaccaaa gccaccgggc aaccaatagc ccttgtcact tttgatcagg
     2461 tagaccgacc ctgaagcgct tttttcgtat tccataaaac ccccttctgt gcgtgagtac
     2521 tcatagtata acaggcgtga gtaccaacgc aagcactaca tgctgaaatc tggcccgccc
     2581 ctgtccatgc ctcgctggcg gggtgccggt gcccgtgcca gctcggcccg cgcaagctgg
     2641 acgctgggca gacccatgac cttgctgacg gtgcgctcga tgtaatccgc ttcgtggccg
     2701 ggcttgcgct ctgccagcgc tgggctggcc tcggccatgg ccttgccgat ttcctcggca
     2761 ctgcggcccc ggctggccag cttctgcgcg gcgataaagt cgcacttgct gaggtcatca
     2821 ccgaagcgct tgaccagccc ggccatctcg ctgcggtact cgtccagcgc cgtgcgccgg
     2881 tggcggctaa gctgccgctc gggcagttcg aggctggcca gcctgcgggc cttctcctgc
     2941 tgccgctggg cctgctcgat ctgctggcca gcctgctgca ccagcgccgg gccagcggtg
     3001 gcggtcttgc ccttggattc acgcagcagc acccacggct gataaccggc gcgggtggtg
     3061 tgcttgtcct tgcggttggt gaagcccgcc aagcggccat agtggcggct gtcggcgctg
     3121 gccgggtcgg cgtcgtactc gctggccagc gtccgggcaa tctgcccccg aagttcaccg
     3181 cctgcggcgt cggccacctt gacccatgcc tgatagttct tcgggctggt ttccactacc
     3241 agggcaggct cccggccctc ggctttcatg tcatccaggt caaactcgct gaggtcgtcc
     3301 accagcacca gaccatgccg ctcctgctcg gcgggcctga tatacacgtc attgccctgg
     3361 gcattcatcc gcttgagcca tggcgtgttc tggagcactt cggcggctga ccattcccgg
     3421 ttcatcatct ggccggtggt ggcgtccctg acgccgatat cgaagcgctc acagcccatg
     3481 gccttgagct gtcggcctat ggcctgcaaa gtcctgtcgt tcttcatcgg gccaccaagc
     3541 gcagccagat cgagccgtcc tcggttgtca gtggcgtcag gtcgagcaag agcaacgatg
     3601 cgatcagcag caccaccgta ggcatcatgg aagccagcat cacggttagc catagcttcc
     3661 agtgccaccc ccgcgacgcg ctccgggcgc tctgcgcggc gctgctcacc tcggcggcta
     3721 cctcccgcaa ctctttggcc agctccaccc atgccgcccc tgtctggcgc tgggctttca
     3781 gccactccgc cgcctgcgcc tcgctggcct gctgggtctg gctcatgacc tgccgggctt
     3841 cgtcggccag tgtcgccatg ctctgggcca gcggttcgat ctgctccgct aactcgttga
     3901 tgcctctgga tttcttcact ctgtcgattg cgttcatggt ctattgcctc ccggtattcc
     3961 tgtaagtcga tgatctgggc gttggcggtg tcgatgttca gggccacgtc tgcccggtcg
     4021 gtgcggatgc cccggccttc catctccacc acgttcggcc ccaggtgaac accgggcagg
     4081 cgctcgatgc cctgcgcctc aagtgttctg tggtcaatgc gggcgtcgtg gccagcccgc
     4141 tctaatgccc ggttggcatg gtcggcccat gcctcgcggg tctgctcaag ccatgccttg
     4201 ggcttgagcg cttcggtctt ctgtgccccg cccttctccg gggtcttgcc gttgtaccgc
     4261 ttgaaccact gagcggcggg ccgctcgatg ccgtcattga tccgctcgga gatcatcagg
     4321 tggcagtgcg ggttctcgcc gccaccggca tggatggcca gcgtatacgg caggcgctcg
     4381 gcaccggtca ggtgctgggc gaactcggac gccagcgcct tctgctggtc gagggtcagc
     4441 tcgaccggca gggcaaattc gacctccttg aacagccgcc cattggcgcg ttcatacagg
     4501 tcggcagcat cccagtagtc ggcgggccgc tcgacgaact ccggcatgtg cccggattcg
     4561 gcgtgcaaga cttcatccat gtcgcgggca tacttgcctt cgcgctggat gtagtcggcc
     4621 ttggccctgg ccgattggcc gcccgacctg ctgccggttt tcgccgtaag gtgataaatc
     4681 gccatgctgc ctcgctgttg cttttgcttt tcggctccat gcaatggccc tcggagagcg
     4741 caccgcccga agggtggccg ttaggccagt ttctcgaaga gaaaccggta agtgcgccct
     4801 cccctacaaa gtagggtcgg gattgccgcc gctgtgcctc catgatagcc tacgagacag
     4861 cacattaaca atggggtgtc aagatggtta aggggagcaa caaggcggcg gatcggctgg
     4921 ccaagctcga agaacaacga gcgcgaatca atgccgaaat tcagcgggtg cgggcaaggg
     4981 aacagcagca agagcgcaag aacgaaacaa ggcgcaaggt gctggtgggg gccatgattt
     5041 tggccaaggt gaacagcagc gagtggccgg aggatcggct catggcggca atggatgcgt
     5101 accttgaacg cgaccacgac cgcgccttgt tcggtctgcc gccacgccag aaggatgagc
     5161 cgggctgaat gatcgaccga gacaggccct gcggggctgc acacgcgccc ccacccttcg
     5221 ggtaggggga aaggccgcta aagcggctaa aagcgctcca gcgtatttct gcggggtttg
     5281 gtgtggggtt tagcgggctt tgcccgcctt tccccctgcc gcgcagcggt ggggcggtgt
     5341 gtagcctagc gcagcgaata gaccagctat ccggcctctg gccgggcata ttgggcaagg
     5401 gcagcagcgc cccacaaggg cgctgataac cgcgcctagt ggattattct tagataatca
     5461 tggatggatt tttccaacac cccgccagcc cccgcccctg ctgggtttgc aggtttgggg
     5521 gcgtgacagt tattgcaggg gttcgtgaca gttattgcag gggggcgtga cagttattgc
     5581 aggggttcgt gacagttagt acgggagtga cgggcactgg ctggcaatgt ctagcaacgg
     5641 caggcatttc ggctgagggt aaaagaactt tccgctaagc gatagactgt atgtaaacac
     5701 agtattgcaa ggacgcggaa catgcctcat gtggcggcca ggacggccag ccgggatcgg
     5761 gatactggtc gttaccagag ccaccgaccc gagcaaaccc ttctctatca gatcgttgac
     5821 gagtattacc cggcattcgc tgcgcttatg gcagagcagg gaaaggaatt gccgggctat
     5881 gtgcaacggg aatttgaaga atttctccaa tgcgggcggc tggagcatgg ctttctacgg
     5941 gttcgctgcg agtcttgcca cgccgagcac ctggtcgctt tcagaaatca atctaaagta
     6001 tatatgagta aacttggtct gacaggcccc tgaattcgca tctagatggt agagccacaa
     6061 acagccggta caagcaacga tctccaggac catctgaatc atgcgcggat gacacgaact
     6121 cacgacggcg atcacagaca ttaacccaca gtacagacac tgcgacaacg tggcaattcg
     6181 tcgcaatacc gtctcactga actggccgat aattgcagac gaacgggtga aacaaaacgg
     6241 ttgacaacat gaagtaaaca cggtacgatg taccacatga aacgacagtg agtcaataca
     6301 gaaacagagg agatattaca tatggtcttc acactcgaag atttcgttgg ggactggcga
     6361 cagacagccg gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt
     6421 cagaatctcg gggtgtccgt aactccgatc caaaggattg tcctgagcgg tgaaaatggg
     6481 ctgaagatcg acatccatgt catcatcccg tatgaaggtc tgagcggcga ccaaatgggc
     6541 cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc
     6601 ctgcactatg gcacactggt aatcgacggg gttacgccga acatgatcga ctatttcgga
     6661 cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg
     6721 tggaacggca acaaaattat cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc
     6781 cgagtaacca tcaacggagt gaccggctgg cggctgtgcg aacgcattct ggcgtaaatc
     6841 cgtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg
     6901 agtttaggga aagagcattt gtcagctgga tcagcagatg ggaaccggat actcctcagc
     6961 ctccagagta gccaatgaga cgccgagcgg ccgcttattt gccgactacc ttggtgatct
     7021 cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt
     7081 cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca
     7141 ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc
     7201 tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg
     7261 gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg
     7321 gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg
     7381 tcagcaagat agccagatca atgtcgatcg tggctggctc gaagataccc gcaagaatgt
     7441 cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga
     7501 tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg
     7561 aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta
     7621 cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg
     7681 agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta
     7741 cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcataatg tttaactt
//
