LOCUS       sequence-208082-        8054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK assembled broad-host-range plasmid encoding GFP driven by CP6 
            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: pBTK621
FEATURES             Location/Qualifiers
     source          1..8054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(36..851)
                     /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(36..851)
                     /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     759..778
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     843..862
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1026..1877)
                     /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(1864..2703)
                     /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(3214..4185)
                     /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            5424..5511
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      5853..6247
                     /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      7694..7741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      7930..8001
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8017..8044
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 gcggccgcgt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
       61 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
      121 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
      181 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
      241 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
      301 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
      361 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg aaatacgcga
      421 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
      481 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
      541 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
      601 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
      661 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
      721 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
      781 tataaatcag catccatgtt ggaatttaat cgcggcctgg agcaagacgt ttcccgttga
      841 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
      901 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
      961 tgttgaataa atcgaacttt tgctgagttg aaggatcagt catgaccaaa atcccttaac
     1021 gtgagtcagc ctgccgcctt gggccgggtg atgtcgtact tgcccgccgc gaactcggtt
     1081 accgtccagc ccagcgcgac cagctccggc aacgcctcgc gcacccgctt gcggcgcttg
     1141 cgcatggtcg aaccactggc ctctgacggc cagacatagc cgcacaaggt atctatggaa
     1201 gccttgccgg ttttgccggg gtcgatccag ccacacagcc gctggtgcag caggcgggcg
     1261 gtttcgctgt ccagcgcccg cacctcgtcc atgctgatgc gcacatgctg gccgccaccc
     1321 atgacggcct gcgcgatcaa ggggttcagg gccacgtaca ggcgcccgtc cgcctcgtcg
     1381 ctggcgtact ccgacagcag ccgaaacccc tgccgcttgc ggccattctg ggcgatgatg
     1441 gataccttcc aaaggcgctc gatgcagtcc tgtatgtgct tgagcgcccc accactatcg
     1501 acctctgccc cgatttcctt tgccagcgcc cgatagctac ctttgaccac atggcattca
     1561 gcggtgacgg cctcccactt gggttccagg aacagccgga gctgccgtcc gccttcggtc
     1621 ttgggttccg ggccaagcac taggccatta ggcccagcca tggccaccag cccttgcagg
     1681 atgcgcagat catcagcgcc cagcggctcc gggccgctga actcgatccg cttgccgtcg
     1741 ccgtagtcat acgtcacgtc cagcttgctg cgcttgcgct cgccccgctt gagggcacgg
     1801 aacaggccgg gggccagaca gtgcgccggg tcgtgccgga cgtggctgag gctgtgcttg
     1861 ttcttaggct tcaccacggg gcaccccctt gctcttgcgc tgcctctcca gcacggcggg
     1921 cttgagcacc ccgccgtcat gccgcctgaa ccaccgatca gcgaacggtg cgccatagtt
     1981 ggccttgctc acaccgaagc ggacgaagaa ccggcgctgg tcgtcgtcca caccccattc
     2041 ctcggcctcg gcgctggtca tgctcgacag gtaggactgc cagcggatgt tatcgaccag
     2101 taccgagctg ccccggctgg cctgctgctg gtcgcctgcg cccatcatgg ccgcgccctt
     2161 gctggcatgg tgcaggaaca cgatagagca cccggtatcg gcggcgatgg cctccatgcg
     2221 accgatgacc tgggccatgg ggccgctggc gttttcttcc tcgatgtgga accggcgcag
     2281 cgtgtccagc accatcaggc ggcggccctc ggcggcgcgc ttgaggccgt cgaaccactc
     2341 cggggccatg atgttgggca ggctgccgat cagcggctgg atcagcaggc cgtcagccac
     2401 ggcttgccgt tcctcggcgc tgaggtgcgc cccaagggcg tgcaggcggt gatgaatggc
     2461 ggtgggcggg tcttcggcgg gcaggtagat caccgggccg gtgggcagtt cgcccacctc
     2521 cagcagatcc ggcccgcctg caatctgtgc ggccagttgc agggccagca tggatttacc
     2581 ggcaccaccg ggcgacacca gcgccccgac cgtaccggcc accatgttgg gcaaaacgta
     2641 gtccagcggt ggcggcgctg ctgcgaacgc ctccagaata ttgataggct tatgggtagc
     2701 cattgattgc ctcctttgca ggcagttggt ggttaggcgc tggcggggtc actacccccg
     2761 ccctgcgccg ctctgagttc ttccaggcac tcgcgcagcg cctcgtattc gtcgtcggtc
     2821 agccagaact tgcgctgacg catccctttg gccttcatgc gctcggcata tcgcgcttgg
     2881 cgtacagcgt cagggctggc cagcaggtcg ccggtctgct tgtccttttg gtctttcata
     2941 tcagtcaccg agaaacttgc cggggccgaa aggcttgtct tcgcggaaca aggacaaggt
     3001 gcagccgtca aggttaaggc tggccatatc agcgactgaa aagcggccag cctcggcctt
     3061 gtttgacgta taaccaaagc caccgggcaa ccaatagccc ttgtcacttt tgatcaggta
     3121 gaccgaccct gaagcgcttt tttcgtattc cataaaaccc ccttctgtgc gtgagtactc
     3181 atagtataac aggcgtgagt accaacgcaa gcactacatg ctgaaatctg gcccgcccct
     3241 gtccatgcct cgctggcggg gtgccggtgc ccgtgccagc tcggcccgcg caagctggac
     3301 gctgggcaga cccatgacct tgctgacggt gcgctcgatg taatccgctt cgtggccggg
     3361 cttgcgctct gccagcgctg ggctggcctc ggccatggcc ttgccgattt cctcggcact
     3421 gcggccccgg ctggccagct tctgcgcggc gataaagtcg cacttgctga ggtcatgacc
     3481 gaagcgcttg accagcccgg ccatctcgct gcggtactcg tccagcgccg tgcgccggtg
     3541 gcggctaagc tgccgctcgg gcagttcgag gctggccagc ctgcgggcct tctcctgctg
     3601 ccgctgggcc tgctcgatct gctggccagc ctgctgcacc agcgccgggc cagcggtggc
     3661 ggtcttgccc ttggattcac gcagcagcac ccacggctga taaccggcgc gggtggtgtg
     3721 cttgtccttg cggttggtga agcccgccaa gcggccatag tggcggctgt cggcgctggc
     3781 cgggtcggcg tcgtactcgc tggccagcgt ccgggcaatc tgcccccgaa gttcaccgcc
     3841 tgcggcgtcg gccaccttga cccatgcctg atagttcttc gggctggttt ccactaccag
     3901 ggcaggctcc cggccctcgg ctttcatgtc atccaggtca aactcgctga ggtcgtccac
     3961 cagcaccaga ccatgccgct cctgctcggc gggcctgata tacacgtcat tgccctgggc
     4021 attcatccgc ttgagccatg gcgtgttctg gagcacttcg gcggctgacc attcccggtt
     4081 catcatctgg ccggtgggtg cgtccctgac gccgatatcg aagcgctcac agcccatggc
     4141 cttgagctgt cggcctatgg cctgcaaagt cctgtcgttc ttcatcgggc caccaagcgc
     4201 agccagatcg agccgtcctc ggttgtcagt ggcgtcaggt cgagcaagag caacgatgcg
     4261 atcagcagca ccaccgtagg catcatggaa gccagcatca cggttagcca tagcttccag
     4321 tgccaccccc gcgacgcgct ccgggcgctc tgcgcggcgc tgctcacctc ggcggctacc
     4381 tcccgcaact ctttggccag ctccacccat gccgcccctg tctggcgctg ggctttcagc
     4441 cactccgccg cctgcgcctc gctggcctgc ttggtctggc tcatgacctg ccgggcttcg
     4501 tcggccagtg tcgccatgct ctgggccagc ggttcgatct gctccgctaa ctcgttgatg
     4561 cctctggatt tcttcactct gtcgattgcg ttcatggtct attgcctccc ggtattcctg
     4621 taagtcgatg atctgggcgt tggcggtgtc gatgttcagg gccacgtctg cccggtcggt
     4681 gcggatgccc cggccttcca tctccaccac gttcggcccc aggtgaacac cgggcaggcg
     4741 ctcgatgccc tgcgcctcaa gtgttctgtg gtcaatgcgg gcgtcgtggc cagcccgctc
     4801 taatgcccgg ttggcatggt cggcccatgc ctcgcgggtc tgctcaagcc atgccttggg
     4861 cttgagcgct tcggtcttct gtgccccgcc cttctccggg gtcttgccgt tgtaccgctt
     4921 gaaccactga gcggcgggcc gctcgatgcc gtcattgatc cgctcggaga tcatcaggtg
     4981 gcagtgcggg ttctcgccgc caccggcatg gatggccagc gtatacggca ggcgctcggc
     5041 accggtcagg tgctgggcga actcggacgc cagcgccttc tgctggtcga gggtcagctc
     5101 gaccggcagg gcaaattcga cctccttgaa cagccgccca ttggcgcgtt catacaggtc
     5161 ggcagcatcc cagtagtcgg cgggccgctc gacgaactcc ggcatgtgcc cggattcggc
     5221 gtgcaagact tcatccatgt cgcgggcata cttgccttcg cgctggatgt agtcggcctt
     5281 ggccctggcc gattggccgc ccgacctgct gccggttttc gccgtaaggt gataaatcgc
     5341 catgctgcct cgctgttgct tttgcttttc ggctccatgc aatggccctc ggagagcgca
     5401 ccgcccgaag ggtggccgtt aggccagttt ctcgaagaga aaccggtaag tgcgccctcc
     5461 cctacaaagt agggtcggga ttgccgccgc tgtgcctcca tgatagccta cgagacagca
     5521 cattaacaat ggggtgtcaa gatggttaag gggagcaaca aggcggcgga tcggctggcc
     5581 aagctcgaag aacaacgagc gcgaatcaat gccgaaattc agcgggagcg ggcaagggaa
     5641 cagcagcaag agcgcaagaa cgaaacaagg cgcaaggtgc tggtgggggc catgattttg
     5701 gccaaggtga acagcagcga gtggccggag gatcggctca tggcggcaat ggatgcgtac
     5761 cttgaacgcg accacgaccg cgccttgttc ggtctgccgc cacgccagaa ggatgagccg
     5821 ggctgaatga tcgaccgaga caggccctgc ggggctgcac acgcgccccc acccttcggg
     5881 tagggggaaa ggccgctaaa gcggctaaaa gcgctccagc gtatttctgc ggggtttggt
     5941 gtggggttta gcgggctttg cccgcctttc cccctgccgc gcagcggtgg ggcggtgtgt
     6001 agcctagcgc agcgaataga ccagctatcc ggcctctggc cgggcatatt gggcaagggc
     6061 agcagcgccc cacaagggcg ctgataaccg cgcctagtgg attattctta gataatcatg
     6121 gatggatttt tccaacaccc cgccagcccc cgcccctgct gggtttgcag gtttgggggc
     6181 gtgacagtta ttgcaggggt tcgtgacagt tattgcaggg gggcgtgaca gttattgcag
     6241 gggttcgtga cagttagtac gggagtgacg ggcactggct ggcaatgtct agcaacggca
     6301 ggcatttcgg ctgagggtaa aagaactttc cgctaagcga tagactgtat gtaaacacag
     6361 tattgcaagg acgcggaaca tgcctcatgt ggcggccagg acggccagcc gggatcggga
     6421 tactggtcgt taccagagcc accgacccga gcaaaccctt ctctatcaga tcgttgacga
     6481 gtattacccg gcattcgctg cgcttatggc agagcaggga aaggaattgc cgggctatgt
     6541 gcaacgggaa tttgaagaat ttctccaatg cgggcggctg gagcatggct ttctacgggt
     6601 tcgctgcgag tcttgccacg ccgagcacct ggtcgctttc agaaatcaat ctaaagtata
     6661 tatgagtaaa cttggtctga caggcccctg aattcgcatc tagatggtag agccacaaac
     6721 agccggtaca agcaacgatc tccaggacca tctgaatcat gcgcggatga cacgaactca
     6781 cgacggcgat cacagacatt aacccacagt acagacactg cgacaacgtg gcaattcgtc
     6841 gcaataccgt ctcactgaac tggccgataa ttgcagacga acgcatgtgg gagtttattc
     6901 ttgacacaga tatttccgga tgatataata actgagtact gttatacaga aacagaggag
     6961 atattacata tgagtaaagg agaagagctt ttcacaggag ttgtcccaat cctcgtggaa
     7021 ttagacggtg atgttaatgg gcacaagttc tctgtcagtg gagagggtga aggcgacgca
     7081 acatatggca agctgaccct taaatttatt tgcaccacgg gtaaactacc tgttccatgg
     7141 ccaacactgg tcactacgtt cgggtatggg gttcagtgct ttgcgcgcta cccagatcac
     7201 atgaaacagc acgacttttt caagagtgca atgcccgaag gctatgtaca ggagagaacc
     7261 atctttttta aggatgacgg caactataag acacgcgccg aagtgaagtt cgagggtgat
     7321 acccttgtta atagaatcga gttaaagggt attgacttta aggaagatgg aaatatttta
     7381 ggccacaaac tggaatataa ctataactcc cataatgtgt acattatggc cgacaagcaa
     7441 aagaacggta tcaaggttaa cttcaagatc agacacaaca ttgaggatgg aagcgttcaa
     7501 ctagccgacc attaccaaca aaacacccca attggcgatg ggcctgtgct gttaccagac
     7561 aaccattacc tgtccactca atctgccctt tcgaaagatc ccaacgaaaa gcgcgaccac
     7621 atggtccttc ttgagtttgt cacggctgct gggattacac acggcatgga tgaactatac
     7681 aaataaatcc taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     7741 gctggctgag catgagacgg aaatctgctc gtcagtggtg ctcacactga cgaatcatgt
     7801 acagatcata ccgatgactg cctggcgact cacaactaag caagacagcc ggaaccagcg
     7861 ccggcgaaca ccactgcata tatggcatat cacaacagtc caactagtgc actgcagtac
     7921 accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     7981 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     8041 tgcgtttata ccga
//
