LOCUS       sequence-208074-        7798 bp    DNA     circular SYN 14-APR-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 Apr 14, 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"
     oriT            2090..2177
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      2519..2913
                     /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             3646..4161
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             complement(4319..5110)
                     /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(5490..6341)
                     /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(6328..7167)
                     /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(join(7678..7798,1..851))
                     /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"
ORIGIN
        1 tcgatgtaat ccgcttcgtg gccgggcttg cgctctgcca gcgctgggct ggcctcggcc
       61 atggccttgc cgatttcctc ggcactgcgg ccccggctgg ccagcttctg cgcggcgata
      121 aagtcgcact tgctgaggtc atgaccgaag cgcttgacca gcccggccat ctcgctgcgg
      181 tactcgtcca gcgccgtgcg ccggtggcgg ctaagctgcc gctcgggcag ttcgaggctg
      241 gccagcctgc gggccttctc ctgctgccgc tgggcctgct cgatctgctg gccagcctgc
      301 tgcaccagcg ccgggccagc ggtggcggtc ttgcccttgg attcacgcag cagcacccac
      361 ggctgataac cggcgcgggt ggtgtgcttg tccttgcggt tggtgaagcc cgccaagcgg
      421 ccatagtggc ggctgtcggc gctggccggg tcggcgtcgt actcgctggc cagcgtccgg
      481 gcaatctgcc cccgaagttc accgcctgcg gcgtcggcca ccttgaccca tgcctgatag
      541 ttcttcgggc tggtttccac taccagggca ggctcccggc cctcggcttt catgtcatcc
      601 aggtcaaact cgctgaggtc gtccaccagc accagaccat gccgctcctg ctcggcgggc
      661 ctgatataca cgtcattgcc ctgggcattc atccgcttga gccatggcgt gttctggagc
      721 acttcggcgg ctgaccattc ccggttcatc atctggccgg tgggtgcgtc cctgacgccg
      781 atatcgaagc gctcacagcc catggccttg agctgtcggc ctatggcctg caaagtcctg
      841 tcgttcttca tcgggccacc aagcgcagcc agatcgagcc gtcctcggtt gtcagtggcg
      901 tcaggtcgag caagagcaac gatgcgatca gcagcaccac cgtaggcatc atggaagcca
      961 gcatcacggt tagccatagc ttccagtgcc acccccgcga cgcgctccgg gcgctctgcg
     1021 cggcgctgct cacctcggcg gctacctccc gcaactcttt ggccagctcc acccatgccg
     1081 cccctgtctg gcgctgggct ttcagccact ccgccgcctg cgcctcgctg gcctgcttgg
     1141 tctggctcat gacctgccgg gcttcgtcgg ccagtgtcgc catgctctgg gccagcggtt
     1201 cgatctgctc cgctaactcg ttgatgcctc tggatttctt cactctgtcg attgcgttca
     1261 tggtctattg cctcccggta ttcctgtaag tcgatgatct gggcgttggc ggtgtcgatg
     1321 ttcagggcca cgtctgcccg gtcggtgcgg atgccccggc cttccatctc caccacgttc
     1381 ggccccaggt gaacaccggg caggcgctcg atgccctgcg cctcaagtgt tctgtggtca
     1441 atgcgggcgt cgtggccagc ccgctctaat gcccggttgg catggtcggc ccatgcctcg
     1501 cgggtctgct caagccatgc cttgggcttg agcgcttcgg tcttctgtgc cccgcccttc
     1561 tccggggtct tgccgttgta ccgcttgaac cactgagcgg cgggccgctc gatgccgtca
     1621 ttgatccgct cggagatcat caggtggcag tgcgggttct cgccgccacc ggcatggatg
     1681 gccagcgtat acggcaggcg ctcggcaccg gtcaggtgct gggcgaactc ggacgccagc
     1741 gccttctgct ggtcgagggt cagctcgacc ggcagggcaa attcgacctc cttgaacagc
     1801 cgcccattgg cgcgttcata caggtcggca gcatcccagt agtcggcggg ccgctcgacg
     1861 aactccggca tgtgcccgga ttcggcgtgc aagacttcat ccatgtcgcg ggcatacttg
     1921 ccttcgcgct ggatgtagtc ggccttggcc ctggccgatt ggccgcccga cctgctgccg
     1981 gttttcgccg taaggtgata aatcgccatg ctgcctcgct gttgcttttg cttttcggct
     2041 ccatgcaatg gccctcggag agcgcaccgc ccgaagggtg gccgttaggc cagtttctcg
     2101 aagagaaacc ggtaagtgcg ccctccccta caaagtaggg tcgggattgc cgccgctgtg
     2161 cctccatgat agcctacgag acagcacatt aacaatgggg tgtcaagatg gttaagggga
     2221 gcaacaaggc ggcggatcgg ctggccaagc tcgaagaaca acgagcgcga atcaatgccg
     2281 aaattcagcg ggagcgggca agggaacagc agcaagagcg caagaacgaa acaaggcgca
     2341 aggtgctggt gggggccatg attttggcca aggtgaacag cagcgagtgg ccggaggatc
     2401 ggctcatggc ggcaatggat gcgtaccttg aacgcgacca cgaccgcgcc ttgttcggtc
     2461 tgccgccacg ccagaaggat gagccgggct gaatgatcga ccgagacagg ccctgcgggg
     2521 ctgcacacgc gcccccaccc ttcgggtagg gggaaaggcc gctaaagcgg ctaaaagcgc
     2581 tccagcgtat ttctgcgggg tttggtgtgg ggtttagcgg gctttgcccg cctttccccc
     2641 tgccgcgcag cggtggggcg gtgtgtagcc tagcgcagcg aatagaccag ctatccggcc
     2701 tctggccggg catattgggc aagggcagca gcgccccaca agggcgctga taaccgcgcc
     2761 tagtggatta ttcttagata atcatggatg gatttttcca acaccccgcc agcccccgcc
     2821 cctgctgggt ttgcaggttt gggggcgtga cagttattgc aggggttcgt gacagttatt
     2881 gcaggggggc gtgacagtta ttgcaggggt tcgtgacagt tagtacggga gtgacgggca
     2941 ctggctggca atgtctagca acggcaggca tttcggctga gggtaaaaga actttccgct
     3001 aagcgataga ctgtatgtaa acacagtatt gcaaggacgc ggaacatgcc tcatgtggcg
     3061 gccaggacgg ccagccggga tcgggatact ggtcgttacc agagccaccg acccgagcaa
     3121 acccttctct atcagatcgt tgacgagtat tacccggcat tcgctgcgct tatggcagag
     3181 cagggaaagg aattgccggg ctatgtgcaa cgggaatttg aagaatttct ccaatgcggg
     3241 cggctggagc atggctttct acgggttcgc tgcgagtctt gccacgccga gcacctggtc
     3301 gctttcagaa atcaatctaa agtatatatg agtaaacttg gtctgacagg cccctgaatt
     3361 cgcatctaga tggtagagcc acaaacagcc ggtacaagca acgatctcca ggaccatctg
     3421 aatcatgcgc ggatgacacg aactcacgac ggcgatcaca gacattaacc cacagtacag
     3481 acactgcgac aacgtggcaa ttcgtcgcaa taccgtctca ctgaactggc cgataattgc
     3541 agacgaacgg gtgaaacaaa acggttgaca acatgaagta aacacggtac gatgtaccac
     3601 atgaaacgac agtgagtcaa tacagaaaca gaggagatat tacatatggt cttcacactc
     3661 gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa
     3721 cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg
     3781 attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa
     3841 ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg
     3901 gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg
     3961 ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa
     4021 aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc
     4081 aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg
     4141 tgcgaacgca ttctggcgta aatccgtaat cgttaatccg caaataacgt aaaaacccgc
     4201 ttcggcgggt ttttttatgg ggggagttta gggaaagagc atttgtcagc tggatcagca
     4261 gatgggaacc ggatactcct cagcctccag agtagccaat gagacgccga gcggccgctt
     4321 atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat
     4381 ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta
     4441 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
     4501 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
     4561 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
     4621 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
     4681 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
     4741 gctcgaagat acccgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
     4801 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
     4861 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
     4921 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
     4981 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
     5041 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
     5101 cttccctcat aatgtttaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     5161 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     5221 gcatagactg taccccaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt
     5281 taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc
     5341 attacagctt acgaaccgaa caggcttatg tccactgggt tcgtgccttc atccgtttcc
     5401 acggtgtgcg tcacccggca accttgggta gcagcgaagt cgaggcattt ctgtcctggc
     5461 tggtcatgac caaaatccct taacgtgagt cagcctgccg ccttgggccg ggtgatgtcg
     5521 tacttgcccg ccgcgaactc ggttaccgtc cagcccagcg cgaccagctc cggcaacgcc
     5581 tcgcgcaccc gcttgcggcg cttgcgcatg gtcgaaccac tggcctctga cggccagaca
     5641 tagccgcaca aggtatctat ggaagccttg ccggttttgc cggggtcgat ccagccacac
     5701 agccgctggt gcagcaggcg ggcggtttcg ctgtccagcg cccgcacctc gtccatgctg
     5761 atgcgcacat gctggccgcc acccatgacg gcctgcgcga tcaaggggtt cagggccacg
     5821 tacaggcgcc cgtccgcctc gtcgctggcg tactccgaca gcagccgaaa cccctgccgc
     5881 ttgcggccat tctgggcgat gatggatacc ttccaaaggc gctcgatgca gtcctgtatg
     5941 tgcttgagcg ccccaccact atcgacctct gccccgattt cctttgccag cgcccgatag
     6001 ctacctttga ccacatggca ttcagcggtg acggcctccc acttgggttc caggaacagc
     6061 cggagctgcc gtccgccttc ggtcttgggt tccgggccaa gcactaggcc attaggccca
     6121 gccatggcca ccagcccttg caggatgcgc agatcatcag cgcccagcgg ctccgggccg
     6181 ctgaactcga tccgcttgcc gtcgccgtag tcatacgtca cgtccagctt gctgcgcttg
     6241 cgctcgcccc gcttgagggc acggaacagg ccgggggcca gacagtgcgc cgggtcgtgc
     6301 cggacgtggc tgaggctgtg cttgttctta ggcttcacca cggggcaccc ccttgctctt
     6361 gcgctgcctc tccagcacgg cgggcttgag caccccgccg tcatgccgcc tgaaccaccg
     6421 atcagcgaac ggtgcgccat agttggcctt gctcacaccg aagcggacga agaaccggcg
     6481 ctggtcgtcg tccacacccc attcctcggc ctcggcgctg gtcatgctcg acaggtagga
     6541 ctgccagcgg atgttatcga ccagtaccga gctgccccgg ctggcctgct gctggtcgcc
     6601 tgcgcccatc atggccgcgc ccttgctggc atggtgcagg aacacgatag agcacccggt
     6661 atcggcggcg atggcctcca tgcgaccgat gacctgggcc atggggccgc tggcgttttc
     6721 ttcctcgatg tggaaccggc gcagcgtgtc cagcaccatc aggcggcggc cctcggcggc
     6781 gcgcttgagg ccgtcgaacc actccggggc catgatgttg ggcaggctgc cgatcagcgg
     6841 ctggatcagc aggccgtcag ccacggcttg ccgttcctcg gcgctgaggt gcgccccaag
     6901 ggcgtgcagg cggtgatgaa tggcggtggg cgggtcttcg gcgggcaggt agatcaccgg
     6961 gccggtgggc agttcgccca cctccagcag atccggcccg cctgcaatct gtgcggccag
     7021 ttgcagggcc agcatggatt taccggcacc accgggcgac accagcgccc cgaccgtacc
     7081 ggccaccatg ttgggcaaaa cgtagtccag cggtggcggc gctgctgcga acgcctccag
     7141 aatattgata ggcttatggg tagccattga ttgcctcctt tgcaggcagt tggtggttag
     7201 gcgctggcgg ggtcactacc cccgccctgc gccgctctga gttcttccag gcactcgcgc
     7261 agcgcctcgt attcgtcgtc ggtcagccag aacttgcgct gacgcatccc tttggccttc
     7321 atgcgctcgg catatcgcgc ttggcgtaca gcgtcagggc tggccagcag gtcgccggtc
     7381 tgcttgtcct tttggtcttt catatcagtc accgagaaac ttgccggggc cgaaaggctt
     7441 gtcttcgcgg aacaaggaca aggtgcagcc gtcaaggtta aggctggcca tatcagcgac
     7501 tgaaaagcgg ccagcctcgg ccttgtttga cgtataacca aagccaccgg gcaaccaata
     7561 gcccttgtca cttttgatca ggtagaccga ccctgaagcg cttttttcgt attccataaa
     7621 acccccttct gtgcgtgagt actcatagta taacaggcgt gagtaccaac gcaagcacta
     7681 catgctgaaa tctggcccgc ccctgtccat gcctcgctgg cggggtgccg gtgcccgtgc
     7741 cagctcggcc cgcgcaagct ggacgctggg cagacccatg accttgctga cggtgcgc
//
