LOCUS       sequence-215545-        8801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK assembled plasmid- broad-host-range GFP expression from PA1 
            promoter (ampicillin resistance).
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: pBTK501
FEATURES             Location/Qualifiers
     source          1..8801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1649..1695)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1704..2327)
                     /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     2235..2254
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2367..2386)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2368..2386)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(2497..3288,3289..3357))
                     /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     3120..3139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3358..3462)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(3515..4366)
                     /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(4353..5192)
                     /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(5703..6674)
                     /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            7913..8000
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      8342..8736
                     /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 gagggtaaaa gaactttccg ctaagcgata gactgtatgt aaacacagta ttgcaaggac
       61 gcggaacatg cctcatgtgg cggccaggac ggccagccgg gatcgggata ctggtcgtta
      121 ccagagccac cgacccgagc aaacccttct ctatcagatc gttgacgagt attacccggc
      181 attcgctgcg cttatggcag agcagggaaa ggaattgccg ggctatgtgc aacgggaatt
      241 tgaagaattt ctccaatgcg ggcggctgga gcatggcttt ctacgggttc gctgcgagtc
      301 ttgccacgcc gagcacctgg tcgctttcag aaatcaatct aaagtatata tgagtaaact
      361 tggtctgaca ggcccctgaa ttcgcatcta gactgatgag acgtggtaga gccacaaaca
      421 gccggtacaa gcaacgatct ccaggaccat ctgaatcatg cgcggatgac acgaactcac
      481 gacggcgatc acagacatta acccacagta cagacactgc gacaacgtgg caattcgtcg
      541 caatacaacg ttatcaaaaa gagtattgac ttaaagtcta acctatagga tacttacagc
      601 catcgagagg gacacggcga atacagaaac agaggagata ttacatatga gtaaaggaga
      661 agagcttttc acaggagttg tcccaatcct cgtggaatta gacggtgatg ttaatgggca
      721 caagttctct gtcagtggag agggtgaagg cgacgcaaca tatggcaagc tgacccttaa
      781 atttatttgc accacgggta aactacctgt tccatggcca acactggtca ctacgttcgg
      841 gtatggggtt cagtgctttg cgcgctaccc agatcacatg aaacagcacg actttttcaa
      901 gagtgcaatg cccgaaggct atgtacagga gagaaccatc ttttttaagg atgacggcaa
      961 ctataagaca cgcgccgaag tgaagttcga gggtgatacc cttgttaata gaatcgagtt
     1021 aaagggtatt gactttaagg aagatggaaa tattttaggc cacaaactgg aatataacta
     1081 taactcccat aatgtgtaca ttatggccga caagcaaaag aacggtatca aggttaactt
     1141 caagatcaga cacaacattg aggatggaag cgttcaacta gccgaccatt accaacaaaa
     1201 caccccaatt ggcgatgggc ctgtgctgtt accagacaac cattacctgt ccactcaatc
     1261 tgccctttcg aaagatccca acgaaaagcg cgaccacatg gtccttcttg agtttgtcac
     1321 ggctgctggg attacacacg gcatggatga actatacaaa taaatccgta atcgttaatc
     1381 cgcaaataac gtaaaaaccc gcttcggcgg gtttttttat ggggggagtt tagggaaaga
     1441 gcatttgtca gctggaaatc tgctcgtcag tggtgctcac actgacgaat catgtacaga
     1501 tcataccgat gactgcctgg cgactcacaa ctaagcaaga cagccggaac cagcgccggc
     1561 gaacaccact gcatatatgg catatcacaa cagtccacgt ctcaagcagt tacagagatg
     1621 ttacgaacca ctagtgcact gcagtacaca aaaaacccct caagacccgt ttagaggccc
     1681 caaggggtta tgctaatgct gctttaagac ccactttcac atttaagttg tttttctaat
     1741 ccgcatatga tcaattcaag gccgaataag aaggctggct ctgcaccttg gtgatcaaat
     1801 aattcgatag cttgtcgtaa taatggcggc atactatcag tagtaggtgt ttccctttct
     1861 tctttagcga cttgatgctc ttgatcttcc aatacgcaac ctaaagtaaa atgccccact
     1921 gcgctgagtg catataatgc attctctagt gaaaaacctt gttggcataa aaaggctaat
     1981 tgattttcga gagtttcata ctgtttttct gtaggccgtg tacctaaatg tacttttgct
     2041 ccatcgcgat gacttagtaa agcacatcta aaacttttag cgttattacg taaaaaatct
     2101 tgccagcttt ccccttctaa agggcaaaag tgagtatggt gcctatctaa catctcaatg
     2161 gctaaggcgt cgagcaaagc ccgcttattt tttacatgcc aatacaatgt aggctgctct
     2221 acacctagct tctgggcgag tttacgggtt gttaaacctt cgattccgac ctcattaagc
     2281 agctctaatg cgctgttaat cactttactt ttatctaaac gagacatgag tatatctcct
     2341 tctacaaaat tatttctaga tctctcccct atagtgagtc gtattaattt ccgagcggcc
     2401 gcgattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2461 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2521 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2581 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcggga
     2641 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2701 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2761 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2821 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2881 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2941 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3001 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3061 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3121 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3181 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3241 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3301 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3361 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3421 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3481 gccacctgtc atgaccaaaa tcccttaacg tgagtcagcc tgccgccttg ggccgggtga
     3541 tgtcgtactt gcccgccgcg aactcggtta ccgtccagcc cagcgcgacc agctccggca
     3601 acgcctcgcg cacccgcttg cggcgcttgc gcatggtcga accactggcc tctgacggcc
     3661 agacatagcc gcacaaggta tctatggaag ccttgccggt tttgccgggg tcgatccagc
     3721 cacacagccg ctggtgcagc aggcgggcgg tttcgctgtc cagcgcccgc acctcgtcca
     3781 tgctgatgcg cacatgctgg ccgccaccca tgacggcctg cgcgatcaag gggttcaggg
     3841 ccacgtacag gcgcccgtcc gcctcgtcgc tggcgtactc cgacagcagc cgaaacccct
     3901 gccgcttgcg gccattctgg gcgatgatgg ataccttcca aaggcgctcg atgcagtcct
     3961 gtatgtgctt gagcgcccca ccactatcga cctctgcccc gatttccttt gccagcgccc
     4021 gatagctacc tttgaccaca tggcattcag cggtgacggc ctcccacttg ggttccagga
     4081 acagccggag ctgccgtccg ccttcggtct tgggttccgg gccaagcact aggccattag
     4141 gcccagccat ggccaccagc ccttgcagga tgcgcagatc atcagcgccc agcggctccg
     4201 ggccgctgaa ctcgatccgc ttgccgtcgc cgtagtcata cgtcacgtcc agcttgctgc
     4261 gcttgcgctc gccccgcttg agggcacgga acaggccggg ggccagacag tgcgccgggt
     4321 cgtgccggac gtggctgagg ctgtgcttgt tcttaggctt caccacgggg cacccccttg
     4381 ctcttgcgct gcctctccag cacggcgggc ttgagcaccc cgccgtcatg ccgcctgaac
     4441 caccgatcag cgaacggtgc gccatagttg gccttgctca caccgaagcg gacgaagaac
     4501 cggcgctggt cgtcgtccac accccattcc tcggcctcgg cgctggtcat gctcgacagg
     4561 taggactgcc agcggatgtt atcgaccagt accgagctgc cccggctggc ctgctgctgg
     4621 tcgcctgcgc ccatcatggc cgcgcccttg ctggcatggt gcaggaacac gatagagcac
     4681 ccggtatcgg cggcgatggc ctccatgcga ccgatgacct gggccatggg gccgctggcg
     4741 ttttcttcct cgatgtggaa ccggcgcagc gtgtccagca ccatcaggcg gcggccctcg
     4801 gcggcgcgct tgaggccgtc gaaccactcc ggggccatga tgttgggcag gctgccgatc
     4861 agcggctgga tcagcaggcc gtcagccacg gcttgccgtt cctcggcgct gaggtgcgcc
     4921 ccaagggcgt gcaggcggtg atgaatggcg gtgggcgggt cttcggcggg caggtagatc
     4981 accgggccgg tgggcagttc gcccacctcc agcagatccg gcccgcctgc aatctgtgcg
     5041 gccagttgca gggccagcat ggatttaccg gcaccaccgg gcgacaccag cgccccgacc
     5101 gtaccggcca ccatgttggg caaaacgtag tccagcggtg gcggcgctgc tgcgaacgcc
     5161 tccagaatat tgataggctt atgggtagcc attgattgcc tcctttgcag gcagttggtg
     5221 gttaggcgct ggcggggtca ctacccccgc cctgcgccgc tctgagttct tccaggcact
     5281 cgcgcagcgc ctcgtattcg tcgtcggtca gccagaactt gcgctgacgc atccctttgg
     5341 ccttcatgcg ctcggcatat cgcgcttggc gtacagcgtc agggctggcc agcaggtcgc
     5401 cggtctgctt gtccttttgg tctttcatat cagtcaccga gaaacttgcc ggggccgaaa
     5461 ggcttgtctt cgcggaacaa ggacaaggtg cagccgtcaa ggttaaggct ggccatatca
     5521 gcgactgaaa agcggccagc ctcggccttg tttgacgtat aaccaaagcc accgggcaac
     5581 caatagccct tgtcactttt gatcaggtag accgaccctg aagcgctttt ttcgtattcc
     5641 ataaaacccc cttctgtgcg tgagtactca tagtataaca ggcgtgagta ccaacgcaag
     5701 cactacatgc tgaaatctgg cccgcccctg tccatgcctc gctggcgggg tgccggtgcc
     5761 cgtgccagct cggcccgcgc aagctggacg ctgggcagac ccatgacctt gctgacggtg
     5821 cgctcgatgt aatccgcttc gtggccgggc ttgcgctctg ccagcgctgg gctggcctcg
     5881 gccatggcct tgccgatttc ctcggcactg cggccccggc tggccagctt ctgcgcggcg
     5941 ataaagtcgc acttgctgag gtcatgaccg aagcgcttga ccagcccggc catctcgctg
     6001 cggtactcgt ccagcgccgt gcgccggtgg cggctaagct gccgctcggg cagttcgagg
     6061 ctggccagcc tgcgggcctt ctcctgctgc cgctgggcct gctcgatctg ctggccagcc
     6121 tgctgcacca gcgccgggcc agcggtggcg gtcttgccct tggattcacg cagcagcacc
     6181 cacggctgat aaccggcgcg ggtggtgtgc ttgtccttgc ggttggtgaa gcccgccaag
     6241 cggccatagt ggcggctgtc ggcgctggcc gggtcggcgt cgtactcgct ggccagcgtc
     6301 cgggcaatct gcccccgaag ttcaccgcct gcggcgtcgg ccaccttgac ccatgcctga
     6361 tagttcttcg ggctggtttc cactaccagg gcaggctccc ggccctcggc tttcatgtca
     6421 tccaggtcaa actcgctgag gtcgtccacc agcaccagac catgccgctc ctgctcggcg
     6481 ggcctgatat acacgtcatt gccctgggca ttcatccgct tgagccatgg cgtgttctgg
     6541 agcacttcgg cggctgacca ttcccggttc atcatctggc cggtgggtgc gtccctgacg
     6601 ccgatatcga agcgctcaca gcccatggcc ttgagctgtc ggcctatggc ctgcaaagtc
     6661 ctgtcgttct tcatcgggcc accaagcgca gccagatcga gccgtcctcg gttgtcagtg
     6721 gcgtcaggtc gagcaagagc aacgatgcga tcagcagcac caccgtaggc atcatggaag
     6781 ccagcatcac ggttagccat agcttccagt gccacccccg cgacgcgctc cgggcgctct
     6841 gcgcggcgct gctcacctcg gcggctacct cccgcaactc tttggccagc tccacccatg
     6901 ccgcccctgt ctggcgctgg gctttcagcc actccgccgc ctgcgcctcg ctggcctgct
     6961 tggtctggct catgacctgc cgggcttcgt cggccagtgt cgccatgctc tgggccagcg
     7021 gttcgatctg ctccgctaac tcgttgatgc ctctggattt cttcactctg tcgattgcgt
     7081 tcatggtcta ttgcctcccg gtattcctgt aagtcgatga tctgggcgtt ggcggtgtcg
     7141 atgttcaggg ccacgtctgc ccggtcggtg cggatgcccc ggccttccat ctccaccacg
     7201 ttcggcccca ggtgaacacc gggcaggcgc tcgatgccct gcgcctcaag tgttctgtgg
     7261 tcaatgcggg cgtcgtggcc agcccgctct aatgcccggt tggcatggtc ggcccatgcc
     7321 tcgcgggtct gctcaagcca tgccttgggc ttgagcgctt cggtcttctg tgccccgccc
     7381 ttctccgggg tcttgccgtt gtaccgcttg aaccactgag cggcgggccg ctcgatgccg
     7441 tcattgatcc gctcggagat catcaggtgg cagtgcgggt tctcgccgcc accggcatgg
     7501 atggccagcg tatacggcag gcgctcggca ccggtcaggt gctgggcgaa ctcggacgcc
     7561 agcgccttct gctggtcgag ggtcagctcg accggcaggg caaattcgac ctccttgaac
     7621 agccgcccat tggcgcgttc atacaggtcg gcagcatccc agtagtcggc gggccgctcg
     7681 acgaactccg gcatgtgccc ggattcggcg tgcaagactt catccatgtc gcgggcatac
     7741 ttgccttcgc gctggatgta gtcggccttg gccctggccg attggccgcc cgacctgctg
     7801 ccggttttcg ccgtaaggtg ataaatcgcc atgctgcctc gctgttgctt ttgcttttcg
     7861 gctccatgca atggccctcg gagagcgcac cgcccgaagg gtggccgtta ggccagtttc
     7921 tcgaagagaa accggtaagt gcgccctccc ctacaaagta gggtcgggat tgccgccgct
     7981 gtgcctccat gatagcctac gagacagcac attaacaatg gggtgtcaag atggttaagg
     8041 ggagcaacaa ggcggcggat cggctggcca agctcgaaga acaacgagcg cgaatcaatg
     8101 ccgaaattca gcgggagcgg gcaagggaac agcagcaaga gcgcaagaac gaaacaaggc
     8161 gcaaggtgct ggtgggggcc atgattttgg ccaaggtgaa cagcagcgag tggccggagg
     8221 atcggctcat ggcggcaatg gatgcgtacc ttgaacgcga ccacgaccgc gccttgttcg
     8281 gtctgccgcc acgccagaag gatgagccgg gctgaatgat cgaccgagac aggccctgcg
     8341 gggctgcaca cgcgccccca cccttcgggt agggggaaag gccgctaaag cggctaaaag
     8401 cgctccagcg tatttctgcg gggtttggtg tggggtttag cgggctttgc ccgcctttcc
     8461 ccctgccgcg cagcggtggg gcggtgtgta gcctagcgca gcgaatagac cagctatccg
     8521 gcctctggcc gggcatattg ggcaagggca gcagcgcccc acaagggcgc tgataaccgc
     8581 gcctagtgga ttattcttag ataatcatgg atggattttt ccaacacccc gccagccccc
     8641 gcccctgctg ggtttgcagg tttgggggcg tgacagttat tgcaggggtt cgtgacagtt
     8701 attgcagggg ggcgtgacag ttattgcagg ggttcgtgac agttagtacg ggagtgacgg
     8761 gcactggctg gcaatgtcta gcaacggcag gcatttcggc t
//
