LOCUS       sequence-216992-        7439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Assembled plasmid- ready for Stage 2 assembly (ConLS' and ConRE'
            connectors with spectinomycin resistance).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7439)
  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 7439)
  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: pBTK527a
FEATURES             Location/Qualifiers
     source          1..7439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      875..1269
                     /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)"
     oriT            1611..1698
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2937..3908
                     /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"
     CDS             4419..5258
                     /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             5245..6096
                     /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             6476..7267
                     /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"
ORIGIN
        1 gatggctcac gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggtta
       61 taactcgcga taatgtcggg caatcaggtg cgatgacaat ctctaatcga ttgtatggga
      121 agcccgttac ataggctcac gcgattaaat tccaacatgg atgctgattt atatgggtat
      181 aaatgggtta taactcgcga taatgtcggg caatcaggtg cgatgacaat ctctaatcga
      241 ttgtatggga agcccgttac gttgtattgc gacgaattgc cacgttgtcg cagtgtctgt
      301 actgtgggtt aatgtctgtg atcgccgtcg tgagttcgtg tcatccgcgc atgattcaga
      361 tggtcctgga gatcgttgct tgtaccggct gtttgtggct ctaccacgtc tcatcagtct
      421 agatgcgaat tcaggggcct gtcagaccaa gtttactcat atatacttta gattgatttc
      481 tgaaagcgac caggtgctcg gcgtggcaag actcgcagcg aacccgtaga aagccatgct
      541 ccagccgccc gcattggaga aattcttcaa attcccgttg cacatagccc ggcaattcct
      601 ttccctgctc tgccataagc gcagcgaatg ccgggtaata ctcgtcaacg atctgataga
      661 gaagggtttg ctcgggtcgg tggctctggt aacgaccagt atcccgatcc cggctggccg
      721 tcctggccgc cacatgaggc atgttccgcg tccttgcaat actgtgttta catacagtct
      781 atcgcttagc ggaaagttct tttaccctca gccgaaatgc ctgccgttgc tagacattgc
      841 cagccagtgc ccgtcactcc cgtactaact gtcacgaacc cctgcaataa ctgtcacgcc
      901 cccctgcaat aactgtcacg aacccctgca ataactgtca cgcccccaaa cctgcaaacc
      961 cagcaggggc gggggctggc ggggtgttgg aaaaatccat ccatgattat ctaagaataa
     1021 tccactaggc gcggttatca gcgcccttgt ggggcgctgc tgcccttgcc caatatgccc
     1081 ggccagaggc cggatagctg gtctattcgc tgcgctaggc tacacaccgc cccaccgctg
     1141 cgcggcaggg ggaaaggcgg gcaaagcccg ctaaacccca caccaaaccc cgcagaaata
     1201 cgctggagcg cttttagccg ctttagcggc ctttccccct acccgaaggg tgggggcgcg
     1261 tgtgcagccc cgcagggcct gtctcggtcg atcattcagc ccggctcatc cttctggcgt
     1321 ggcggcagac cgaacaaggc gcggtcgtgg tcgcgttcaa ggtacgcatc cattgccgcc
     1381 atgagccgat cctccggcca ctcgctgctg ttcaccttgg ccaaaatcat ggcccccacc
     1441 agcaccttgc gccttgtttc gttcttgcgc tcttgctgct gttcccttgc ccgcacccgc
     1501 tgaatttcgg cattgattcg cgctcgttgt tcttcgagct tggccagccg atccgccgcc
     1561 ttgttgctcc ccttaaccat cttgacaccc cattgttaat gtgctgtctc gtaggctatc
     1621 atggaggcac agcggcggca atcccgaccc tactttgtag gggagggcgc acttaccggt
     1681 ttctcttcga gaaactggcc taacggccac ccttcgggcg gtgcgctctc cgagggccat
     1741 tgcatggagc cgaaaagcaa aagcaacagc gaggcagcat ggcgatttat caccttacgg
     1801 cgaaaaccgg cagcaggtcg ggcggccaat cggccagggc caaggccgac tacatccagc
     1861 gcgaaggcaa gtatgcccgc gacatggatg aagtcttgca cgccgaatcc gggcacatgc
     1921 cggagttcgt cgagcggccc gccgactact gggatgctgc cgacctgtat gaacgcgcca
     1981 atgggcggct gttcaaggag gtcgaatttg ccctgccggt cgagctgacc ctcgaccagc
     2041 agaaggcgct ggcgtccgag ttcgcccagc acctgaccgg tgccgagcgc ctgccgtata
     2101 cgctggccat ccatgccggt ggcggcgaga acccgcactg ccacctgatg atctccgagc
     2161 ggatcaatga cggcatcgag cggcccgccg ctcagtggtt caagcggtac aacggcaaga
     2221 ccccggagaa gggcggggca cagaagaccg aagcgctcaa gcccaaggca tggcttgagc
     2281 agacccgcga ggcatgggcc gaccatgcca accgggcatt agagcgggct ggccacgacg
     2341 cccgcattga ccacagaaca cttgaggcgc agggcatcga gcgcctgccc ggtgttcacc
     2401 tggggccgaa cgtggtggag atggaaggcc ggggcatccg caccgaccgg gcagacgtgg
     2461 ccctgaacat cgacaccgcc aacgcccaga tcatcgactt acaggaatac cgggaggcaa
     2521 tagaccatga acgcaatcga cagagtgaag aaatccagag gcatcaacga gttagcggag
     2581 cagatcgaac cgctggccca gagcatggcg acactggccg acgaagcccg gcaggtcatg
     2641 agccagaccc agcaggccag cgaggcgcag gcggcggagt ggctgaaagc ccagcgccag
     2701 acaggggcgg catgggtgga gctggccaaa gagttgcggg aggtagccgc cgaggtgagc
     2761 agcgccgcgc agagcgcccg gagcgcgtcg cgggggtggc actggaagct atggctaacc
     2821 gtgatgctgg cttccatgat gcctacggtg gtgctgctga tcgcatcgtt gctcttgctc
     2881 gacctgacgc cactgacaac cgaggacggc tcgatctggc tgcgcttggt ggcccgatga
     2941 agaacgacag gactttgcag gccataggcc gacagctcaa ggccatgggc tgtgagcgct
     3001 tcgatatcgg cgtcagggac gccaccaccg gccagatgat gaaccgggaa tggtcagccg
     3061 ccgaagtgct ccagaacacg ccatggctca agcggatgaa tgcccagggc aatgacgtgt
     3121 atatcaggcc cgccgagcag gagcggcatg gtctggtgct ggtggacgac ctcagcgagt
     3181 ttgacctgga tgacatgaaa gccgagggcc gggagcctgc cctggtagtg gaaaccagcc
     3241 cgaagaacta tcaggcatgg gtcaaggtgg ccgacgccgc aggcggtgaa cttcgggggc
     3301 agattgcccg gacgctggcc agcgagtacg acgccgaccc ggccagcgcc gacagccgcc
     3361 actatggccg cttggcgggc ttcaccaacc gcaaggacaa gcacaccacc cgcgccggtt
     3421 atcagccgtg ggtgctgctg cgtgaatcca agggcaagac cgccaccgct ggcccggcgc
     3481 tggtgcagca ggctggccag cagatcgagc aggcccagcg gcagcaggag aaggcccgca
     3541 ggctggccag cctcgaactg cccgagcggc agcttagccg ccaccggcgc acggcgctgg
     3601 acgagtaccg cagcgagatg gccgggctgg tcaagcgctt cggtgatgac ctcagcaagt
     3661 gcgactttat cgccgcgcag aagctggcca gccggggccg cagtgccgag gaaatcggca
     3721 aggccatggc cgaggccagc ccagcgctgg cagagcgcaa gcccggccac gaagcggatt
     3781 acatcgagcg caccgtcagc aaggtcatgg gtctgcccag cgtccagctt gcgcgggccg
     3841 agctggcacg ggcaccggca ccccgccagc gaggcatgga caggggcggg ccagatttca
     3901 gcatgtagtg cttgcgttgg tactcacgcc tgttatacta tgagtactca cgcacagaag
     3961 ggggttttat ggaatacgaa aaaagcgctt cagggtcggt ctacctgatc aaaagtgaca
     4021 agggctattg gttgcccggt ggctttggtt atacgtcaaa caaggccgag gctggccgct
     4081 tttcagtcgc tgatatggcc agccttaacc ttgacggctg caccttgtcc ttgttccgcg
     4141 aagacaagcc tttcggcccc ggcaagtttc tcggtgactg atatgaaaga ccaaaaggac
     4201 aagcagaccg gcgacctgct ggccagccct gacgctgtac gccaagcgcg atatgccgag
     4261 cgcatgaagg ccaaagggat gcgtcagcgc aagttctggc tgaccgacga cgaatacgag
     4321 gcgctgcgcg agtgcctgga agaactcaga gcggcgcagg gcgggggtag tgaccccgcc
     4381 agcgcctaac caccaactgc ctgcaaagga ggcaatcaat ggctacccat aagcctatca
     4441 atattctgga ggcgttcgca gcagcgccgc caccgctgga ctacgttttg cccaacatgg
     4501 tggccggtac ggtcggggcg ctggtgtcgc ccggtggtgc cggtaaatcc atgctggccc
     4561 tgcaactggc cgcacagatt gcaggcgggc cggatctgct ggaggtgggc gaactgccca
     4621 ccggcccggt gatctacctg cccgccgaag acccgcccac cgccattcat caccgcctgc
     4681 acgcccttgg ggcgcacctc agcgccgagg aacggcaagc cgtggctgac ggcctgctga
     4741 tccagccgct gatcggcagc ctgcccaaca tcatggcccc ggagtggttc gacggcctca
     4801 agcgcgccgc cgagggccgc cgcctgatgg tgctggacac gctgcgccgg ttccacatcg
     4861 aggaagaaaa cgccagcggc cccatggccc aggtcatcgg tcgcatggag gccatcgccg
     4921 ccgataccgg gtgctctatc gtgttcctgc accatgccag caagggcgcg gccatgatgg
     4981 gcgcaggcga ccagcagcag gccagccggg gcagctcggt actggtcgat aacatccgct
     5041 ggcagtccta cctgtcgagc atgaccagcg ccgaggccga ggaatggggt gtggacgacg
     5101 accagcgccg gttcttcgtc cgcttcggtg tgagcaaggc caactatggc gcaccgttcg
     5161 ctgatcggtg gttcaggcgg catgacggcg gggtgctcaa gcccgccgtg ctggagaggc
     5221 agcgcaagag caagggggtg ccccgtggtg aagcctaaga acaagcacag cctcagccac
     5281 gtccggcacg acccggcgca ctgtctggcc cccggcctgt tccgtgccct caagcggggc
     5341 gagcgcaagc gcagcaagct ggacgtgacg tatgactacg gcgacggcaa gcggatcgag
     5401 ttcagcggcc cggagccgct gggcgctgat gatctgcgca tcctgcaagg gctggtggcc
     5461 atggctgggc ctaatggcct agtgcttggc ccggaaccca agaccgaagg cggacggcag
     5521 ctccggctgt tcctggaacc caagtgggag gccgtcaccg ctgatgccat ggtggtcaaa
     5581 ggtagctatc gggcgctggc aaaggaaatc ggggcagagg tcgatagtgg tggggcgctc
     5641 aagcacatac aggactgcat cgagcgcctt tggaaggtat ccatcatcgc ccagaatggc
     5701 cgcaagcggc aggggtttcg gctgctgtcg gagtacgcca gcgacgaggc ggacgggcgc
     5761 ctgtacgtgg ccctgaaccc cttgatcgcg caggccgtca tgggtggcgg ccagcatgtg
     5821 cgcatcagca tggacgaggt gcgggcgctg gacagcgaaa ccgcccgcct gctgcaccag
     5881 cggctgtgtg gctggatcga ccccggcaaa accggcaagg cttccataga taccttgtgc
     5941 ggctatgtct ggccgtcaga ggccagtggt tcgaccatgc gcaagcgccg ccagcgggtg
     6001 cgcgaggcgt tgccggagct ggtcgcgctg ggctggacgg taaccgagtt cgcggcgggc
     6061 aagtacgaca tcacccggcc caaggcggca ggctgactca cgttaaggga ttttggtcat
     6121 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     6181 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     6241 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     6301 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     6361 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     6421 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     6481 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     6541 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     6601 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     6661 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     6721 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     6781 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg cgggtatctt
     6841 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     6901 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     6961 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     7021 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     7081 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     7141 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     7201 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     7261 caaataagcg gccgctcggg ttgtaccctc catcgcctcg cattctcgga tgtcctgggg
     7321 gttcgtaaca tctctgtaac tgcttgagac gcagctgaca aatgctcttt ccctaaactc
     7381 cccccataaa aaaacccgcc gaagcgggtt tttacgttat ttgcggatta acgattacg
//
