LOCUS       sequence-208058-        7762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 8 Bacterial origin (broad-host-range) for golden gate 
            assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7762)
  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 7762)
  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: pBTK403
FEATURES             Location/Qualifiers
     source          1..7762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..792)
                     /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"
     CDS             complement(1172..2023)
                     /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(2010..2849)
                     /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(3360..4331)
                     /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            5570..5657
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      5999..6393
                     /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)"
     protein_bind    6845..6863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6870..6888
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             6906..6917
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6924..7601
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      7619..7690
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7706..7733
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
       61 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
      121 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
      181 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
      241 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
      301 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
      361 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
      421 tggctcgaag atacccgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
      481 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
      541 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
      601 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
      661 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
      721 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
      781 cgcttccctc ataatgttta actttgtttt agggcgactg ccctgctgcg taacatcgtt
      841 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
      901 aggcatagac tgtaccccaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc
      961 gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt
     1021 gcattacagc ttacgaaccg aacaggctta tgtccactgg gttcgtgcct tcatccgttt
     1081 ccacggtgtg cgtcacccgg caaccttggg tagcagcgaa gtcgaggcat ttctgtcctg
     1141 gctggtcatg accaaaatcc cttaacgtga gtcagcctgc cgccttgggc cgggtgatgt
     1201 cgtacttgcc cgccgcgaac tcggttaccg tccagcccag cgcgaccagc tccggcaacg
     1261 cctcgcgcac ccgcttgcgg cgcttgcgca tggtcgaacc actggcctct gacggccaga
     1321 catagccgca caaggtatct atggaagcct tgccggtttt gccggggtcg atccagccac
     1381 acagccgctg gtgcagcagg cgggcggttt cgctgtccag cgcccgcacc tcgtccatgc
     1441 tgatgcgcac atgctggccg ccacccatga cggcctgcgc gatcaagggg ttcagggcca
     1501 cgtacaggcg cccgtccgcc tcgtcgctgg cgtactccga cagcagccga aacccctgcc
     1561 gcttgcggcc attctgggcg atgatggata ccttccaaag gcgctcgatg cagtcctgta
     1621 tgtgcttgag cgccccacca ctatcgacct ctgccccgat ttcctttgcc agcgcccgat
     1681 agctaccttt gaccacatgg cattcagcgg tgacggcctc ccacttgggt tccaggaaca
     1741 gccggagctg ccgtccgcct tcggtcttgg gttccgggcc aagcactagg ccattaggcc
     1801 cagccatggc caccagccct tgcaggatgc gcagatcatc agcgcccagc ggctccgggc
     1861 cgctgaactc gatccgcttg ccgtcgccgt agtcatacgt cacgtccagc ttgctgcgct
     1921 tgcgctcgcc ccgcttgagg gcacggaaca ggccgggggc cagacagtgc gccgggtcgt
     1981 gccggacgtg gctgaggctg tgcttgttct taggcttcac cacggggcac ccccttgctc
     2041 ttgcgctgcc tctccagcac ggcgggcttg agcaccccgc cgtcatgccg cctgaaccac
     2101 cgatcagcga acggtgcgcc atagttggcc ttgctcacac cgaagcggac gaagaaccgg
     2161 cgctggtcgt cgtccacacc ccattcctcg gcctcggcgc tggtcatgct cgacaggtag
     2221 gactgccagc ggatgttatc gaccagtacc gagctgcccc ggctggcctg ctgctggtcg
     2281 cctgcgccca tcatggccgc gcccttgctg gcatggtgca ggaacacgat agagcacccg
     2341 gtatcggcgg cgatggcctc catgcgaccg atgacctggg ccatggggcc gctggcgttt
     2401 tcttcctcga tgtggaaccg gcgcagcgtg tccagcacca tcaggcggcg gccctcggcg
     2461 gcgcgcttga ggccgtcgaa ccactccggg gccatgatgt tgggcaggct gccgatcagc
     2521 ggctggatca gcaggccgtc agccacggct tgccgttcct cggcgctgag gtgcgcccca
     2581 agggcgtgca ggcggtgatg aatggcggtg ggcgggtctt cggcgggcag gtagatcacc
     2641 gggccggtgg gcagttcgcc cacctccagc agatccggcc cgcctgcaat ctgtgcggcc
     2701 agttgcaggg ccagcatgga tttaccggca ccaccgggcg acaccagcgc cccgaccgta
     2761 ccggccacca tgttgggcaa aacgtagtcc agcggtggcg gcgctgctgc gaacgcctcc
     2821 agaatattga taggcttatg ggtagccatt gattgcctcc tttgcaggca gttggtggtt
     2881 aggcgctggc ggggtcacta cccccgccct gcgccgctct gagttcttcc aggcactcgc
     2941 gcagcgcctc gtattcgtcg tcggtcagcc agaacttgcg ctgacgcatc cctttggcct
     3001 tcatgcgctc ggcatatcgc gcttggcgta cagcgtcagg gctggccagc aggtcgccgg
     3061 tctgcttgtc cttttggtct ttcatatcag tcaccgagaa acttgccggg gccgaaaggc
     3121 ttgtcttcgc ggaacaagga caaggtgcag ccgtcaaggt taaggctggc catatcagcg
     3181 actgaaaagc ggccagcctc ggccttgttt gacgtataac caaagccacc gggcaaccaa
     3241 tagcccttgt cacttttgat caggtagacc gaccctgaag cgcttttttc gtattccata
     3301 aaaccccctt ctgtgcgtga gtactcatag tataacaggc gtgagtacca acgcaagcac
     3361 tacatgctga aatctggccc gcccctgtcc atgcctcgct ggcggggtgc cggtgcccgt
     3421 gccagctcgg cccgcgcaag ctggacgctg ggcagaccca tgaccttgct gacggtgcgc
     3481 tcgatgtaat ccgcttcgtg gccgggcttg cgctctgcca gcgctgggct ggcctcggcc
     3541 atggccttgc cgatttcctc ggcactgcgg ccccggctgg ccagcttctg cgcggcgata
     3601 aagtcgcact tgctgaggtc atgaccgaag cgcttgacca gcccggccat ctcgctgcgg
     3661 tactcgtcca gcgccgtgcg ccggtggcgg ctaagctgcc gctcgggcag ttcgaggctg
     3721 gccagcctgc gggccttctc ctgctgccgc tgggcctgct cgatctgctg gccagcctgc
     3781 tgcaccagcg ccgggccagc ggtggcggtc ttgcccttgg attcacgcag cagcacccac
     3841 ggctgataac cggcgcgggt ggtgtgcttg tccttgcggt tggtgaagcc cgccaagcgg
     3901 ccatagtggc ggctgtcggc gctggccggg tcggcgtcgt actcgctggc cagcgtccgg
     3961 gcaatctgcc cccgaagttc accgcctgcg gcgtcggcca ccttgaccca tgcctgatag
     4021 ttcttcgggc tggtttccac taccagggca ggctcccggc cctcggcttt catgtcatcc
     4081 aggtcaaact cgctgaggtc gtccaccagc accagaccat gccgctcctg ctcggcgggc
     4141 ctgatataca cgtcattgcc ctgggcattc atccgcttga gccatggcgt gttctggagc
     4201 acttcggcgg ctgaccattc ccggttcatc atctggccgg tgggtgcgtc cctgacgccg
     4261 atatcgaagc gctcacagcc catggccttg agctgtcggc ctatggcctg caaagtcctg
     4321 tcgttcttca tcgggccacc aagcgcagcc agatcgagcc gtcctcggtt gtcagtggcg
     4381 tcaggtcgag caagagcaac gatgcgatca gcagcaccac cgtaggcatc atggaagcca
     4441 gcatcacggt tagccatagc ttccagtgcc acccccgcga cgcgctccgg gcgctctgcg
     4501 cggcgctgct cacctcggcg gctacctccc gcaactcttt ggccagctcc acccatgccg
     4561 cccctgtctg gcgctgggct ttcagccact ccgccgcctg cgcctcgctg gcctgcttgg
     4621 tctggctcat gacctgccgg gcttcgtcgg ccagtgtcgc catgctctgg gccagcggtt
     4681 cgatctgctc cgctaactcg ttgatgcctc tggatttctt cactctgtcg attgcgttca
     4741 tggtctattg cctcccggta ttcctgtaag tcgatgatct gggcgttggc ggtgtcgatg
     4801 ttcagggcca cgtctgcccg gtcggtgcgg atgccccggc cttccatctc caccacgttc
     4861 ggccccaggt gaacaccggg caggcgctcg atgccctgcg cctcaagtgt tctgtggtca
     4921 atgcgggcgt cgtggccagc ccgctctaat gcccggttgg catggtcggc ccatgcctcg
     4981 cgggtctgct caagccatgc cttgggcttg agcgcttcgg tcttctgtgc cccgcccttc
     5041 tccggggtct tgccgttgta ccgcttgaac cactgagcgg cgggccgctc gatgccgtca
     5101 ttgatccgct cggagatcat caggtggcag tgcgggttct cgccgccacc ggcatggatg
     5161 gccagcgtat acggcaggcg ctcggcaccg gtcaggtgct gggcgaactc ggacgccagc
     5221 gccttctgct ggtcgagggt cagctcgacc ggcagggcaa attcgacctc cttgaacagc
     5281 cgcccattgg cgcgttcata caggtcggca gcatcccagt agtcggcggg ccgctcgacg
     5341 aactccggca tgtgcccgga ttcggcgtgc aagacttcat ccatgtcgcg ggcatacttg
     5401 ccttcgcgct ggatgtagtc ggccttggcc ctggccgatt ggccgcccga cctgctgccg
     5461 gttttcgccg taaggtgata aatcgccatg ctgcctcgct gttgcttttg cttttcggct
     5521 ccatgcaatg gccctcggag agcgcaccgc ccgaagggtg gccgttaggc cagtttctcg
     5581 aagagaaacc ggtaagtgcg ccctccccta caaagtaggg tcgggattgc cgccgctgtg
     5641 cctccatgat agcctacgag acagcacatt aacaatgggg tgtcaagatg gttaagggga
     5701 gcaacaaggc ggcggatcgg ctggccaagc tcgaagaaca acgagcgcga atcaatgccg
     5761 aaattcagcg ggagcgggca agggaacagc agcaagagcg caagaacgaa acaaggcgca
     5821 aggtgctggt gggggccatg attttggcca aggtgaacag cagcgagtgg ccggaggatc
     5881 ggctcatggc ggcaatggat gcgtaccttg aacgcgacca cgaccgcgcc ttgttcggtc
     5941 tgccgccacg ccagaaggat gagccgggct gaatgatcga ccgagacagg ccctgcgggg
     6001 ctgcacacgc gcccccaccc ttcgggtagg gggaaaggcc gctaaagcgg ctaaaagcgc
     6061 tccagcgtat ttctgcgggg tttggtgtgg ggtttagcgg gctttgcccg cctttccccc
     6121 tgccgcgcag cggtggggcg gtgtgtagcc tagcgcagcg aatagaccag ctatccggcc
     6181 tctggccggg catattgggc aagggcagca gcgccccaca agggcgctga taaccgcgcc
     6241 tagtggatta ttcttagata atcatggatg gatttttcca acaccccgcc agcccccgcc
     6301 cctgctgggt ttgcaggttt gggggcgtga cagttattgc aggggttcgt gacagttatt
     6361 gcaggggggc gtgacagtta ttgcaggggt tcgtgacagt tagtacggga gtgacgggca
     6421 ctggctggca atgtctagca acggcaggca tttcggctga gggtaaaaga actttccgct
     6481 aagcgataga ctgtatgtaa acacagtatt gcaaggacgc ggaacatgcc tcatgtggcg
     6541 gccaggacgg ccagccggga tcgggatact ggtcgttacc agagccaccg acccgagcaa
     6601 acccttctct atcagatcgt tgacgagtat tacccggcat tcgctgcgct tatggcagag
     6661 cagggaaagg aattgccggg ctatgtgcaa cgggaatttg aagaatttct ccaatgcggg
     6721 cggctggagc atggctttct acgggttcgc tgcgagtctt gccacgccga gcacctggtc
     6781 gctttcagaa atcaatctaa agtatatatg agtaaacttg gtctgacagg ccccttgaga
     6841 ccagtcccta tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacgg
     6901 atctgaaaga ggagaaagga tctatggcga gtagcgaaga cgttatcaaa gagttcatgc
     6961 gtttcaaagt tcgtatggaa ggttccgtta acggtcacga gttcgaaatc gaaggtgaag
     7021 gtgaaggtcg tccgtacgaa ggtactcaga ccgctaaact gaaagttacc aaaggtggtc
     7081 cgctgccgtt cgcttgggac atcctgtccc cgcagttcca gtacggttcc aaagcttacg
     7141 ttaaacaccc ggctgacatc ccggactacc tgaaactgtc cttcccggaa ggtttcaaat
     7201 gggaacgtgt tatgaacttc gaagacggtg gtgttgttac cgttacccag gactcctccc
     7261 tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg tactaacttc ccgtccgacg
     7321 gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt atgtacccgg
     7381 aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac ggtggtcact
     7441 acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg ccgggtgctt
     7501 acaaaaccga catcaaactg gacatcacct cccacaacga agactacacc atcgttgaac
     7561 agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataaggatct ccaggcatca
     7621 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     7681 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttataa
     7741 gtcggtctca ccgagcggcc gc
//
