LOCUS       sequence-208080-        8054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK assembled broad-host-range plasmid encoding GFP driven by CP12b 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8054)
  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 8054)
  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: pBTK619
FEATURES             Location/Qualifiers
     source          1..8054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      130..201
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      217..244
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(290..1105)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(290..1105)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1013..1032
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1097..1116
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1280..2131)
                     /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(2118..2957)
                     /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(3468..4439)
                     /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            5678..5765
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      6107..6501
                     /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)"
     terminator      7948..7995
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 acagatcata ccgatgactg cctggcgact cacaactaag caagacagcc ggaaccagcg
       61 ccggcgaaca ccactgcata tatggcatat cacaacagtc caactagtgc actgcagtac
      121 accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      181 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
      241 tgcgtttata ccgagcggcc gcgtgttaca accaattaac caattctgat tagaaaaact
      301 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
      361 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
      421 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
      481 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
      541 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
      601 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
      661 ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
      721 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
      781 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
      841 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
      901 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
      961 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     1021 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag
     1081 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     1141 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     1201 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagtcatgac
     1261 caaaatccct taacgtgagt cagcctgccg ccttgggccg ggtgatgtcg tacttgcccg
     1321 ccgcgaactc ggttaccgtc cagcccagcg cgaccagctc cggcaacgcc tcgcgcaccc
     1381 gcttgcggcg cttgcgcatg gtcgaaccac tggcctctga cggccagaca tagccgcaca
     1441 aggtatctat ggaagccttg ccggttttgc cggggtcgat ccagccacac agccgctggt
     1501 gcagcaggcg ggcggtttcg ctgtccagcg cccgcacctc gtccatgctg atgcgcacat
     1561 gctggccgcc acccatgacg gcctgcgcga tcaaggggtt cagggccacg tacaggcgcc
     1621 cgtccgcctc gtcgctggcg tactccgaca gcagccgaaa cccctgccgc ttgcggccat
     1681 tctgggcgat gatggatacc ttccaaaggc gctcgatgca gtcctgtatg tgcttgagcg
     1741 ccccaccact atcgacctct gccccgattt cctttgccag cgcccgatag ctacctttga
     1801 ccacatggca ttcagcggtg acggcctccc acttgggttc caggaacagc cggagctgcc
     1861 gtccgccttc ggtcttgggt tccgggccaa gcactaggcc attaggccca gccatggcca
     1921 ccagcccttg caggatgcgc agatcatcag cgcccagcgg ctccgggccg ctgaactcga
     1981 tccgcttgcc gtcgccgtag tcatacgtca cgtccagctt gctgcgcttg cgctcgcccc
     2041 gcttgagggc acggaacagg ccgggggcca gacagtgcgc cgggtcgtgc cggacgtggc
     2101 tgaggctgtg cttgttctta ggcttcacca cggggcaccc ccttgctctt gcgctgcctc
     2161 tccagcacgg cgggcttgag caccccgccg tcatgccgcc tgaaccaccg atcagcgaac
     2221 ggtgcgccat agttggcctt gctcacaccg aagcggacga agaaccggcg ctggtcgtcg
     2281 tccacacccc attcctcggc ctcggcgctg gtcatgctcg acaggtagga ctgccagcgg
     2341 atgttatcga ccagtaccga gctgccccgg ctggcctgct gctggtcgcc tgcgcccatc
     2401 atggccgcgc ccttgctggc atggtgcagg aacacgatag agcacccggt atcggcggcg
     2461 atggcctcca tgcgaccgat gacctgggcc atggggccgc tggcgttttc ttcctcgatg
     2521 tggaaccggc gcagcgtgtc cagcaccatc aggcggcggc cctcggcggc gcgcttgagg
     2581 ccgtcgaacc actccggggc catgatgttg ggcaggctgc cgatcagcgg ctggatcagc
     2641 aggccgtcag ccacggcttg ccgttcctcg gcgctgaggt gcgccccaag ggcgtgcagg
     2701 cggtgatgaa tggcggtggg cgggtcttcg gcgggcaggt agatcaccgg gccggtgggc
     2761 agttcgccca cctccagcag atccggcccg cctgcaatct gtgcggccag ttgcagggcc
     2821 agcatggatt taccggcacc accgggcgac accagcgccc cgaccgtacc ggccaccatg
     2881 ttgggcaaaa cgtagtccag cggtggcggc gctgctgcga acgcctccag aatattgata
     2941 ggcttatggg tagccattga ttgcctcctt tgcaggcagt tggtggttag gcgctggcgg
     3001 ggtcactacc cccgccctgc gccgctctga gttcttccag gcactcgcgc agcgcctcgt
     3061 attcgtcgtc ggtcagccag aacttgcgct gacgcatccc tttggccttc atgcgctcgg
     3121 catatcgcgc ttggcgtaca gcgtcagggc tggccagcag gtcgccggtc tgcttgtcct
     3181 tttggtcttt catatcagtc accgagaaac ttgccggggc cgaaaggctt gtcttcgcgg
     3241 aacaaggaca aggtgcagcc gtcaaggtta aggctggcca tatcagcgac tgaaaagcgg
     3301 ccagcctcgg ccttgtttga cgtataacca aagccaccgg gcaaccaata gcccttgtca
     3361 cttttgatca ggtagaccga ccctgaagcg cttttttcgt attccataaa acccccttct
     3421 gtgcgtgagt actcatagta taacaggcgt gagtaccaac gcaagcacta catgctgaaa
     3481 tctggcccgc ccctgtccat gcctcgctgg cggggtgccg gtgcccgtgc cagctcggcc
     3541 cgcgcaagct ggacgctggg cagacccatg accttgctga cggtgcgctc gatgtaatcc
     3601 gcttcgtggc cgggcttgcg ctctgccagc gctgggctgg cctcggccat ggccttgccg
     3661 atttcctcgg cactgcggcc ccggctggcc agcttctgcg cggcgataaa gtcgcacttg
     3721 ctgaggtcat gaccgaagcg cttgaccagc ccggccatct cgctgcggta ctcgtccagc
     3781 gccgtgcgcc ggtggcggct aagctgccgc tcgggcagtt cgaggctggc cagcctgcgg
     3841 gccttctcct gctgccgctg ggcctgctcg atctgctggc cagcctgctg caccagcgcc
     3901 gggccagcgg tggcggtctt gcccttggat tcacgcagca gcacccacgg ctgataaccg
     3961 gcgcgggtgg tgtgcttgtc cttgcggttg gtgaagcccg ccaagcggcc atagtggcgg
     4021 ctgtcggcgc tggccgggtc ggcgtcgtac tcgctggcca gcgtccgggc aatctgcccc
     4081 cgaagttcac cgcctgcggc gtcggccacc ttgacccatg cctgatagtt cttcgggctg
     4141 gtttccacta ccagggcagg ctcccggccc tcggctttca tgtcatccag gtcaaactcg
     4201 ctgaggtcgt ccaccagcac cagaccatgc cgctcctgct cggcgggcct gatatacacg
     4261 tcattgccct gggcattcat ccgcttgagc catggcgtgt tctggagcac ttcggcggct
     4321 gaccattccc ggttcatcat ctggccggtg ggtgcgtccc tgacgccgat atcgaagcgc
     4381 tcacagccca tggccttgag ctgtcggcct atggcctgca aagtcctgtc gttcttcatc
     4441 gggccaccaa gcgcagccag atcgagccgt cctcggttgt cagtggcgtc aggtcgagca
     4501 agagcaacga tgcgatcagc agcaccaccg taggcatcat ggaagccagc atcacggtta
     4561 gccatagctt ccagtgccac ccccgcgacg cgctccgggc gctctgcgcg gcgctgctca
     4621 cctcggcggc tacctcccgc aactctttgg ccagctccac ccatgccgcc cctgtctggc
     4681 gctgggcttt cagccactcc gccgcctgcg cctcgctggc ctgcttggtc tggctcatga
     4741 cctgccgggc ttcgtcggcc agtgtcgcca tgctctgggc cagcggttcg atctgctccg
     4801 ctaactcgtt gatgcctctg gatttcttca ctctgtcgat tgcgttcatg gtctattgcc
     4861 tcccggtatt cctgtaagtc gatgatctgg gcgttggcgg tgtcgatgtt cagggccacg
     4921 tctgcccggt cggtgcggat gccccggcct tccatctcca ccacgttcgg ccccaggtga
     4981 acaccgggca ggcgctcgat gccctgcgcc tcaagtgttc tgtggtcaat gcgggcgtcg
     5041 tggccagccc gctctaatgc ccggttggca tggtcggccc atgcctcgcg ggtctgctca
     5101 agccatgcct tgggcttgag cgcttcggtc ttctgtgccc cgcccttctc cggggtcttg
     5161 ccgttgtacc gcttgaacca ctgagcggcg ggccgctcga tgccgtcatt gatccgctcg
     5221 gagatcatca ggtggcagtg cgggttctcg ccgccaccgg catggatggc cagcgtatac
     5281 ggcaggcgct cggcaccggt caggtgctgg gcgaactcgg acgccagcgc cttctgctgg
     5341 tcgagggtca gctcgaccgg cagggcaaat tcgacctcct tgaacagccg cccattggcg
     5401 cgttcataca ggtcggcagc atcccagtag tcggcgggcc gctcgacgaa ctccggcatg
     5461 tgcccggatt cggcgtgcaa gacttcatcc atgtcgcggg catacttgcc ttcgcgctgg
     5521 atgtagtcgg ccttggccct ggccgattgg ccgcccgacc tgctgccggt tttcgccgta
     5581 aggtgataaa tcgccatgct gcctcgctgt tgcttttgct tttcggctcc atgcaatggc
     5641 cctcggagag cgcaccgccc gaagggtggc cgttaggcca gtttctcgaa gagaaaccgg
     5701 taagtgcgcc ctcccctaca aagtagggtc gggattgccg ccgctgtgcc tccatgatag
     5761 cctacgagac agcacattaa caatggggtg tcaagatggt taaggggagc aacaaggcgg
     5821 cggatcggct ggccaagctc gaagaacaac gagcgcgaat caatgccgaa attcagcggg
     5881 agcgggcaag ggaacagcag caagagcgca agaacgaaac aaggcgcaag gtgctggtgg
     5941 gggccatgat tttggccaag gtgaacagca gcgagtggcc ggaggatcgg ctcatggcgg
     6001 caatggatgc gtaccttgaa cgcgaccacg accgcgcctt gttcggtctg ccgccacgcc
     6061 agaaggatga gccgggctga atgatcgacc gagacaggcc ctgcggggct gcacacgcgc
     6121 ccccaccctt cgggtagggg gaaaggccgc taaagcggct aaaagcgctc cagcgtattt
     6181 ctgcggggtt tggtgtgggg tttagcgggc tttgcccgcc tttccccctg ccgcgcagcg
     6241 gtggggcggt gtgtagccta gcgcagcgaa tagaccagct atccggcctc tggccgggca
     6301 tattgggcaa gggcagcagc gccccacaag ggcgctgata accgcgccta gtggattatt
     6361 cttagataat catggatgga tttttccaac accccgccag cccccgcccc tgctgggttt
     6421 gcaggtttgg gggcgtgaca gttattgcag gggttcgtga cagttattgc aggggggcgt
     6481 gacagttatt gcaggggttc gtgacagtta gtacgggagt gacgggcact ggctggcaat
     6541 gtctagcaac ggcaggcatt tcggctgagg gtaaaagaac tttccgctaa gcgatagact
     6601 gtatgtaaac acagtattgc aaggacgcgg aacatgcctc atgtggcggc caggacggcc
     6661 agccgggatc gggatactgg tcgttaccag agccaccgac ccgagcaaac ccttctctat
     6721 cagatcgttg acgagtatta cccggcattc gctgcgctta tggcagagca gggaaaggaa
     6781 ttgccgggct atgtgcaacg ggaatttgaa gaatttctcc aatgcgggcg gctggagcat
     6841 ggctttctac gggttcgctg cgagtcttgc cacgccgagc acctggtcgc tttcagaaat
     6901 caatctaaag tatatatgag taaacttggt ctgacaggcc cctgaattcg catctagatg
     6961 gtagagccac aaacagccgg tacaagcaac gatctccagg accatctgaa tcatgcgcgg
     7021 atgacacgaa ctcacgacgg cgatcacaga cattaaccca cagtacagac actgcgacaa
     7081 cgtggcaatt cgtcgcaata ccgtctcact gaactggccg ataattgcag acgaacgcat
     7141 atacaagttt attcttgaca ctagtcggcc aaaatgatat aatacctgag tactgttata
     7201 cagaaacaga ggagatatta catatgagta aaggagaaga gcttttcaca ggagttgtcc
     7261 caatcctcgt ggaattagac ggtgatgtta atgggcacaa gttctctgtc agtggagagg
     7321 gtgaaggcga cgcaacatat ggcaagctga cccttaaatt tatttgcacc acgggtaaac
     7381 tacctgttcc atggccaaca ctggtcacta cgttcgggta tggggttcag tgctttgcgc
     7441 gctacccaga tcacatgaaa cagcacgact ttttcaagag tgcaatgccc gaaggctatg
     7501 tacaggagag aaccatcttt tttaaggatg acggcaacta taagacacgc gccgaagtga
     7561 agttcgaggg tgataccctt gttaatagaa tcgagttaaa gggtattgac tttaaggaag
     7621 atggaaatat tttaggccac aaactggaat ataactataa ctcccataat gtgtacatta
     7681 tggccgacaa gcaaaagaac ggtatcaagg ttaacttcaa gatcagacac aacattgagg
     7741 atggaagcgt tcaactagcc gaccattacc aacaaaacac cccaattggc gatgggcctg
     7801 tgctgttacc agacaaccat tacctgtcca ctcaatctgc cctttcgaaa gatcccaacg
     7861 aaaagcgcga ccacatggtc cttcttgagt ttgtcacggc tgctgggatt acacacggca
     7921 tggatgaact atacaaataa atcctaacta gcataacccc ttggggcctc taaacgggtc
     7981 ttgaggggtt ttttgctggc tgagcatgag acggaaatct gctcgtcagt ggtgctcaca
     8041 ctgacgaatc atgt
//
