LOCUS       sequence-220593-        8801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK assembled broad-host-range plasmid encoding GFP driven by PA3 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8801)
  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 8801)
  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: pBTK510
FEATURES             Location/Qualifiers
     source          1..8801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1445..1491)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1500..2123)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     2031..2050
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2163..2182)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2164..2182)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(2293..3084,3085..3153))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2916..2935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3154..3258)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(3311..4162)
                     /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(4149..4988)
                     /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(5499..6470)
                     /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            7709..7796
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      8138..8532
                     /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)"
ORIGIN
        1 gggaaaggaa ttgccgggct atgtgcaacg ggaatttgaa gaatttctcc aatgcgggcg
       61 gctggagcat ggctttctac gggttcgctg cgagtcttgc cacgccgagc acctggtcgc
      121 tttcagaaat caatctaaag tatatatgag taaacttggt ctgacaggcc cctgaattcg
      181 catctagact gatgagacgt ggtagagcca caaacagccg gtacaagcaa cgatctccag
      241 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      301 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat acaacgggtg aaacaaaacg
      361 gttgacaaca tgaagtaaac acggtacgat gtaccacatg aaacgacagt gagtcaatac
      421 agaaacagag gagatattac atatgagtaa aggagaagag cttttcacag gagttgtccc
      481 aatcctcgtg gaattagacg gtgatgttaa tgggcacaag ttctctgtca gtggagaggg
      541 tgaaggcgac gcaacatatg gcaagctgac ccttaaattt atttgcacca cgggtaaact
      601 acctgttcca tggccaacac tggtcactac gttcgggtat ggggttcagt gctttgcgcg
      661 ctacccagat cacatgaaac agcacgactt tttcaagagt gcaatgcccg aaggctatgt
      721 acaggagaga accatctttt ttaaggatga cggcaactat aagacacgcg ccgaagtgaa
      781 gttcgagggt gatacccttg ttaatagaat cgagttaaag ggtattgact ttaaggaaga
      841 tggaaatatt ttaggccaca aactggaata taactataac tcccataatg tgtacattat
      901 ggccgacaag caaaagaacg gtatcaaggt taacttcaag atcagacaca acattgagga
      961 tggaagcgtt caactagccg accattacca acaaaacacc ccaattggcg atgggcctgt
     1021 gctgttacca gacaaccatt acctgtccac tcaatctgcc ctttcgaaag atcccaacga
     1081 aaagcgcgac cacatggtcc ttcttgagtt tgtcacggct gctgggatta cacacggcat
     1141 ggatgaacta tacaaataaa tccgtaatcg ttaatccgca aataacgtaa aaacccgctt
     1201 cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagctg gaaatctgct
     1261 cgtcagtggt gctcacactg acgaatcatg tacagatcat accgatgact gcctggcgac
     1321 tcacaactaa gcaagacagc cggaaccagc gccggcgaac accactgcat atatggcata
     1381 tcacaacagt ccacgtctca agcagttaca gagatgttac gaaccactag tgcactgcag
     1441 tacacaaaaa acccctcaag acccgtttag aggccccaag gggttatgct aatgctgctt
     1501 taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa ttcaaggccg
     1561 aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg tcgtaataat
     1621 ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg atgctcttga
     1681 tcttccaata cgcaacctaa agtaaaatgc cccactgcgc tgagtgcata taatgcattc
     1741 tctagtgaaa aaccttgttg gcataaaaag gctaattgat tttcgagagt ttcatactgt
     1801 ttttctgtag gccgtgtacc taaatgtact tttgctccat cgcgatgact tagtaaagca
     1861 catctaaaac ttttagcgtt attacgtaaa aaatcttgcc agctttcccc ttctaaaggg
     1921 caaaagtgag tatggtgcct atctaacatc tcaatggcta aggcgtcgag caaagcccgc
     1981 ttatttttta catgccaata caatgtaggc tgctctacac ctagcttctg ggcgagttta
     2041 cgggttgtta aaccttcgat tccgacctca ttaagcagct ctaatgcgct gttaatcact
     2101 ttacttttat ctaaacgaga catgagtata tctccttcta caaaattatt tctagatctc
     2161 tcccctatag tgagtcgtat taatttccga gcggccgcga ttatcaaaaa ggatcttcac
     2221 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2401 accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg ctccagattt
     2461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2581 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2881 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3241 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgtcatga ccaaaatccc
     3301 ttaacgtgag tcagcctgcc gccttgggcc gggtgatgtc gtacttgccc gccgcgaact
     3361 cggttaccgt ccagcccagc gcgaccagct ccggcaacgc ctcgcgcacc cgctggcggc
     3421 gcttgcgcat ggtcgaacca ctggcctctg acggccagac atagccgcac aaggtatcta
     3481 tggaagcctt gccggttttg ccggggtcga tccagccaca cagccgctgg tgcagcaggc
     3541 gggcggtttc gctgtccagc gcccgcacct cgtccatgct gatgcgcaca tgctggccgc
     3601 cacccatgac ggcctgcgcg atcaaggggt tcagggccac gtacaggcgc ccgtccgcct
     3661 cgtcgctggc gtactccgac agcagccgaa acccctgccg cttgcggcca ttctgggcga
     3721 tgatggatac cttccaaagg cgctcgatgc agtcctgtat gtgcttgagc gccccaccac
     3781 tatcgacctc tgccccgatt tcctttgcca gcgcccgata gctacctttg accaccatgg
     3841 catcagcggt gacggcctcc cacttgggtt ccaggaacag ccggagctgc cgtccgcctt
     3901 cggtcttggg ttccgggcca agcactaggc cattaggccc agccatggcc accagccctt
     3961 gcaggatgcg cagatcatca gcgcccagcg gctccgggcc gctgaactcg atccgcttgc
     4021 cgtcgccgta gtcatacgtc acgtccagct tgctgcgctt gcgctcgccc cgcttgaggg
     4081 cacggaacag gccgggggcc agacagtgcg ccgggtcgtg ccggacgtgg ctgaggctgt
     4141 gcttgttctt aggcttcacc acggggcacc cccttgctct tgcgctgcct ctccagcacg
     4201 gcgggcttga gcaccccgcc gtcatgccgc ctgaaccacc gatcagcgaa cggtgcgcca
     4261 tagttggcct tgctcacacc gaagcggacg aagaaccggc gctggtcgtc gtccacaccc
     4321 cattcctcgg cctcggcgct ggtcatgctc gacaggtagg actgccagcg gatgttatcg
     4381 accagtaccg agctgccccg gctggcctgc tgctggtcgc ctgcgcccat catggccgcg
     4441 cccttgctgg catggtgcag gaacacgata gagcacccgg tatcggcggc gatggcctcc
     4501 atgcgaccga tgacctgggc catggggccg ctggcgtttt cttcctcgat gtggaaccgg
     4561 cgcagcgtgt ccagcaccat caggcggcgg ccctcggcgg cgcgcttgag gccgtcgaac
     4621 cactccgggg ccatgatgtt gggcaggctg ccgatcagcg gctggatcag caggccgtca
     4681 gccacggctt gccgttcctc ggcgctgagg tgcgccccaa gggcgtgcag gcggtgatga
     4741 atggcggtgg gcgggtcttc ggcgggcagg tagatcaccg ggccggtggg cagttcgccc
     4801 acctccagca gatccggccc gcctgcaatc tgtgcggcca gttgcagggc cagcatggat
     4861 ttaccggcac caccgggcga caccagcgcc ccgaccgtac cggccaccat gttgggcaaa
     4921 acgtagtcca gcggtggcgg cgctgctgcg aacgcctcca gaatattgat aggcttatgg
     4981 gtagccattg attgcctcct ttgcaggcag ttggtggtta ggcgctggcg gggtcactac
     5041 ccccgccctg cgccgctctg agttcttcca ggcactcgcg cagcgcctcg tattcgtcgt
     5101 cggtcagcca gaacttgcgc tgacgcatcc ctttggcctt catgcgctcg gcatatcgcg
     5161 cttggcgtac agcgtcaggg ctggccagca ggtcgccggt ctgcttgtcc ttttggtctt
     5221 tcatatcagt caccgagaaa cttgccgggg ccgaaaggct tgtcttcgcg gaacaaggac
     5281 aaggtgcagc cgtcaaggtt aaggctggcc atatcagcga ctgaaaagcg gccagcctcg
     5341 gccttgtttg acgtataacc aaagccaccg ggcaaccaat agcccttgtc acttttgatc
     5401 aggtagaccg accctgaagc gcttttttcg tattccataa aacccccttc tgtgcgtgag
     5461 tactcatagt ataacaggcg tgagtaccaa cgcaagcact acatgctgaa atctggcccg
     5521 cccctgtcca tgcctcgctg gcggggtgcc ggtgcccgtg ccagctcggc ccgcgcaagc
     5581 tggacgctgg gcagacccat gaccttgctg acggtgcgct cgatgtaatc cgcttcgtgg
     5641 ccgggcttgc gctctgccag cgctgggctg gcctcggcca tggccttgcc gatttcctcg
     5701 gcactgcggc cccggctggc cagcttctgc gcggcgataa agtcgcactt gctgaggtca
     5761 tcaccgaagc gcttgaccag cccggccatc tcgctgcggt actcgtccag cgccgtgcgc
     5821 cggtggcggc taagctgccg ctcgggcagt tcgaggctgg ccagcctgcg ggccttctcc
     5881 tgctgccgct gggcctgctc gatctgctgg ccagcctgct gcaccagcgc cgggccagcg
     5941 gtggcggtct tgcccttgga ttcacgcagc agcacccacg gctgataacc ggcgcgggtg
     6001 gtgtgcttgt ccttgcggtt ggtgaagccc gccaagcggc catagtggcg gctgtcggcg
     6061 ctggccgggt cggcgtcgta ctcgctggcc agcgtccggg caatctgccc ccgaagttca
     6121 ccgcctgcgg cgtcggccac cttgacccat gcctgatagt tcttcgggct ggtttccact
     6181 accagggcag gctcccggcc ctcggctttc atgtcatcca ggtcaaactc gctgaggtcg
     6241 tccaccagca ccagaccatg ccgctcctgc tcggcgggcc tgatatacac gtcattgccc
     6301 tgggcattca tccgcttgag ccatggcgtg ttctggagca cttcggcggc tgaccattcc
     6361 cggttcatca tctggccggt ggtggcgtcc ctgacgccga tatcgaagcg ctcacagccc
     6421 atggccttga gctgtcggcc tatggcctgc aaagtcctgt cgttcttcat cgggccacca
     6481 agcgcagcca gatcgagccg tcctcggttg tcagtggcgt caggtcgagc aagagcaacg
     6541 atgcgatcag cagcaccacc gtaggcatca tggaagccag catcacggtt agccatagct
     6601 tccagtgcca cccccgcgac gcgctccggg cgctctgcgc ggcgctgctc acctcggcgg
     6661 ctacctcccg caactctttg gccagctcca cccatgccgc ccctgtctgg cgctgggctt
     6721 tcagccactc cgccgcctgc gcctcgctgg cctgctgggt ctggctcatg acctgccggg
     6781 cttcgtcggc cagtgtcgcc atgctctggg ccagcggttc gatctgctcc gctaactcgt
     6841 tgatgcctct ggatttcttc actctgtcga ttgcgttcat ggtctattgc ctcccggtat
     6901 tcctgtaagt cgatgatctg ggcgttggcg gtgtcgatgt tcagggccac gtctgcccgg
     6961 tcggtgcgga tgccccggcc ttccatctcc accacgttcg gccccaggtg aacaccgggc
     7021 aggcgctcga tgccctgcgc ctcaagtgtt ctgtggtcaa tgcgggcgtc gtggccagcc
     7081 cgctctaatg cccggttggc atggtcggcc catgcctcgc gggtctgctc aagccatgcc
     7141 ttgggcttga gcgcttcggt cttctgtgcc ccgcccttct ccggggtctt gccgttgtac
     7201 cgcttgaacc actgagcggc gggccgctcg atgccgtcat tgatccgctc ggagatcatc
     7261 aggtggcagt gcgggttctc gccgccaccg gcatggatgg ccagcgtata cggcaggcgc
     7321 tcggcaccgg tcaggtgctg ggcgaactcg gacgccagcg ccttctgctg gtcgagggtc
     7381 agctcgaccg gcagggcaaa ttcgacctcc ttgaacagcc gcccattggc gcgttcatac
     7441 aggtcggcag catcccagta gtcggcgggc cgctcgacga actccggcat gtgcccggat
     7501 tcggcgtgca agacttcatc catgtcgcgg gcatacttgc cttcgcgctg gatgtagtcg
     7561 gccttggccc tggccgattg gccgcccgac ctgctgccgg ttttcgccgt aaggtgataa
     7621 atcgccatgc tgcctcgctg ttgcttttgc ttttcggctc catgcaatgg ccctcggaga
     7681 gcgcaccgcc cgaagggtgg ccgttaggcc agtttctcga agagaaaccg gtaagtgcgc
     7741 cctcccctac aaagtagggt cgggattgcc gccgctgtgc ctccatgata gcctacgaga
     7801 cagcacatta acaatggggt gtcaagatgg ttaaggggag caacaaggcg gcggatcggc
     7861 tggccaagct cgaagaacaa cgagcgcgaa tcaatgccga aattcagcgg gtgcgggcaa
     7921 gggaacagca gcaagagcgc aagaacgaaa caaggcgcaa ggtgctggtg ggggccatga
     7981 ttttggccaa ggtgaacagc agcgagtggc cggaggatcg gctcatggcg gcaatggatg
     8041 cgtaccttga acgcgaccac gaccgcgcct tgttcggtct gccgccacgc cagaaggatg
     8101 agccgggctg aatgatcgac cgagacaggc cctgcggggc tgcacacgcg cccccaccct
     8161 tcgggtaggg ggaaaggccg ctaaagcggc taaaagcgct ccagcgtatt tctgcggggt
     8221 ttggtgtggg gtttagcggg ctttgcccgc ctttccccct gccgcgcagc ggtggggcgg
     8281 tgtgtagcct agcgcagcga atagaccagc tatccggcct ctggccgggc atattgggca
     8341 agggcagcag cgccccacaa gggcgctgat aaccgcgcct agtggattat tcttagataa
     8401 tcatggatgg atttttccaa caccccgcca gcccccgccc ctgctgggtt tgcaggtttg
     8461 ggggcgtgac agttattgca ggggttcgtg acagttattg caggggggcg tgacagttat
     8521 tgcaggggtt cgtgacagtt agtacgggag tgacgggcac tggctggcaa tgtctagcaa
     8581 cggcaggcat ttcggctgag ggtaaaagaa ctttccgcta agcgatagac tgtatgtaaa
     8641 cacagtattg caaggacgcg gaacatgcct catgtggcgg ccaggacggc cagccgggat
     8701 cgggatactg gtcgttacca gagccaccga cccgagcaaa cccttctcta tcagatcgtt
     8761 gacgagtatt acccggcatt cgctgcgctt atggcagagc a
//
