LOCUS       sequence-216994-        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"
     terminator      13..40
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(70..861)
                     /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(1241..2092)
                     /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(2079..2918)
                     /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(3429..4400)
                     /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            5639..5726
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      6068..6462
                     /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    6914..6932
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6939..6957
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             6975..6986
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6993..7670
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      7688..7759
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataagtc ggtctcaccg
       61 agcggccgct tatttgccga ctaccttggt gatctcgcct ttcacgtagt ggacaaattc
      121 ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat aagcctgtct
      181 agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc
      241 gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag
      301 cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc
      361 atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg
      421 acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc
      481 gatcgtggct ggctcgaaga tacccgcaag aatgtcattg cgctgccatt ctccaaattg
      541 cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac
      601 ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt
      661 gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat
      721 atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa
      781 cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc
      841 ggcgatcacc gcttccctca taatgtttaa ctttgtttta gggcgactgc cctgctgcgt
      901 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
      961 ggatgcccga ggcatagact gtaccccaaa aaaacagtca taacaagcca tgaaaaccgc
     1021 cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat
     1081 acgctacttg cattacagct tacgaaccga acaggcttat gtccactggg ttcgtgcctt
     1141 catccgtttc cacggtgtgc gtcacccggc aaccttgggt agcagcgaag tcgaggcatt
     1201 tctgtcctgg ctggtcatga ccaaaatccc ttaacgtgag tcagcctgcc gccttgggcc
     1261 gggtgatgtc gtacttgccc gccgcgaact cggttaccgt ccagcccagc gcgaccagct
     1321 ccggcaacgc ctcgcgcacc cgctggcggc gcttgcgcat ggtcgaacca ctggcctctg
     1381 acggccagac atagccgcac aaggtatcta tggaagcctt gccggttttg ccggggtcga
     1441 tccagccaca cagccgctgg tgcagcaggc gggcggtttc gctgtccagc gcccgcacct
     1501 cgtccatgct gatgcgcaca tgctggccgc cacccatgac ggcctgcgcg atcaaggggt
     1561 tcagggccac gtacaggcgc ccgtccgcct cgtcgctggc gtactccgac agcagccgaa
     1621 acccctgccg cttgcggcca ttctgggcga tgatggatac cttccaaagg cgctcgatgc
     1681 agtcctgtat gtgcttgagc gccccaccac tatcgacctc tgccccgatt tcctttgcca
     1741 gcgcccgata gctacctttg accaccatgg catcagcggt gacggcctcc cacttgggtt
     1801 ccaggaacag ccggagctgc cgtccgcctt cggtcttggg ttccgggcca agcactaggc
     1861 cattaggccc agccatggcc accagccctt gcaggatgcg cagatcatca gcgcccagcg
     1921 gctccgggcc gctgaactcg atccgcttgc cgtcgccgta gtcatacgtc acgtccagct
     1981 tgctgcgctt gcgctcgccc cgcttgaggg cacggaacag gccgggggcc agacagtgcg
     2041 ccgggtcgtg ccggacgtgg ctgaggctgt gcttgttctt aggcttcacc acggggcacc
     2101 cccttgctct tgcgctgcct ctccagcacg gcgggcttga gcaccccgcc gtcatgccgc
     2161 ctgaaccacc gatcagcgaa cggtgcgcca tagttggcct tgctcacacc gaagcggacg
     2221 aagaaccggc gctggtcgtc gtccacaccc cattcctcgg cctcggcgct ggtcatgctc
     2281 gacaggtagg actgccagcg gatgttatcg accagtaccg agctgccccg gctggcctgc
     2341 tgctggtcgc ctgcgcccat catggccgcg cccttgctgg catggtgcag gaacacgata
     2401 gagcacccgg tatcggcggc gatggcctcc atgcgaccga tgacctgggc catggggccg
     2461 ctggcgtttt cttcctcgat gtggaaccgg cgcagcgtgt ccagcaccat caggcggcgg
     2521 ccctcggcgg cgcgcttgag gccgtcgaac cactccgggg ccatgatgtt gggcaggctg
     2581 ccgatcagcg gctggatcag caggccgtca gccacggctt gccgttcctc ggcgctgagg
     2641 tgcgccccaa gggcgtgcag gcggtgatga atggcggtgg gcgggtcttc ggcgggcagg
     2701 tagatcaccg ggccggtggg cagttcgccc acctccagca gatccggccc gcctgcaatc
     2761 tgtgcggcca gttgcagggc cagcatggat ttaccggcac caccgggcga caccagcgcc
     2821 ccgaccgtac cggccaccat gttgggcaaa acgtagtcca gcggtggcgg cgctgctgcg
     2881 aacgcctcca gaatattgat aggcttatgg gtagccattg attgcctcct ttgcaggcag
     2941 ttggtggtta ggcgctggcg gggtcactac ccccgccctg cgccgctctg agttcttcca
     3001 ggcactcgcg cagcgcctcg tattcgtcgt cggtcagcca gaacttgcgc tgacgcatcc
     3061 ctttggcctt catgcgctcg gcatatcgcg cttggcgtac agcgtcaggg ctggccagca
     3121 ggtcgccggt ctgcttgtcc ttttggtctt tcatatcagt caccgagaaa cttgccgggg
     3181 ccgaaaggct tgtcttcgcg gaacaaggac aaggtgcagc cgtcaaggtt aaggctggcc
     3241 atatcagcga ctgaaaagcg gccagcctcg gccttgtttg acgtataacc aaagccaccg
     3301 ggcaaccaat agcccttgtc acttttgatc aggtagaccg accctgaagc gcttttttcg
     3361 tattccataa aacccccttc tgtgcgtgag tactcatagt ataacaggcg tgagtaccaa
     3421 cgcaagcact acatgctgaa atctggcccg cccctgtcca tgcctcgctg gcggggtgcc
     3481 ggtgcccgtg ccagctcggc ccgcgcaagc tggacgctgg gcagacccat gaccttgctg
     3541 acggtgcgct cgatgtaatc cgcttcgtgg ccgggcttgc gctctgccag cgctgggctg
     3601 gcctcggcca tggccttgcc gatttcctcg gcactgcggc cccggctggc cagcttctgc
     3661 gcggcgataa agtcgcactt gctgaggtca tcaccgaagc gcttgaccag cccggccatc
     3721 tcgctgcggt actcgtccag cgccgtgcgc cggtggcggc taagctgccg ctcgggcagt
     3781 tcgaggctgg ccagcctgcg ggccttctcc tgctgccgct gggcctgctc gatctgctgg
     3841 ccagcctgct gcaccagcgc cgggccagcg gtggcggtct tgcccttgga ttcacgcagc
     3901 agcacccacg gctgataacc ggcgcgggtg gtgtgcttgt ccttgcggtt ggtgaagccc
     3961 gccaagcggc catagtggcg gctgtcggcg ctggccgggt cggcgtcgta ctcgctggcc
     4021 agcgtccggg caatctgccc ccgaagttca ccgcctgcgg cgtcggccac cttgacccat
     4081 gcctgatagt tcttcgggct ggtttccact accagggcag gctcccggcc ctcggctttc
     4141 atgtcatcca ggtcaaactc gctgaggtcg tccaccagca ccagaccatg ccgctcctgc
     4201 tcggcgggcc tgatatacac gtcattgccc tgggcattca tccgcttgag ccatggcgtg
     4261 ttctggagca cttcggcggc tgaccattcc cggttcatca tctggccggt ggtggcgtcc
     4321 ctgacgccga tatcgaagcg ctcacagccc atggccttga gctgtcggcc tatggcctgc
     4381 aaagtcctgt cgttcttcat cgggccacca agcgcagcca gatcgagccg tcctcggttg
     4441 tcagtggcgt caggtcgagc aagagcaacg atgcgatcag cagcaccacc gtaggcatca
     4501 tggaagccag catcacggtt agccatagct tccagtgcca cccccgcgac gcgctccggg
     4561 cgctctgcgc ggcgctgctc acctcggcgg ctacctcccg caactctttg gccagctcca
     4621 cccatgccgc ccctgtctgg cgctgggctt tcagccactc cgccgcctgc gcctcgctgg
     4681 cctgctgggt ctggctcatg acctgccggg cttcgtcggc cagtgtcgcc atgctctggg
     4741 ccagcggttc gatctgctcc gctaactcgt tgatgcctct ggatttcttc actctgtcga
     4801 ttgcgttcat ggtctattgc ctcccggtat tcctgtaagt cgatgatctg ggcgttggcg
     4861 gtgtcgatgt tcagggccac gtctgcccgg tcggtgcgga tgccccggcc ttccatctcc
     4921 accacgttcg gccccaggtg aacaccgggc aggcgctcga tgccctgcgc ctcaagtgtt
     4981 ctgtggtcaa tgcgggcgtc gtggccagcc cgctctaatg cccggttggc atggtcggcc
     5041 catgcctcgc gggtctgctc aagccatgcc ttgggcttga gcgcttcggt cttctgtgcc
     5101 ccgcccttct ccggggtctt gccgttgtac cgcttgaacc actgagcggc gggccgctcg
     5161 atgccgtcat tgatccgctc ggagatcatc aggtggcagt gcgggttctc gccgccaccg
     5221 gcatggatgg ccagcgtata cggcaggcgc tcggcaccgg tcaggtgctg ggcgaactcg
     5281 gacgccagcg ccttctgctg gtcgagggtc agctcgaccg gcagggcaaa ttcgacctcc
     5341 ttgaacagcc gcccattggc gcgttcatac aggtcggcag catcccagta gtcggcgggc
     5401 cgctcgacga actccggcat gtgcccggat tcggcgtgca agacttcatc catgtcgcgg
     5461 gcatacttgc cttcgcgctg gatgtagtcg gccttggccc tggccgattg gccgcccgac
     5521 ctgctgccgg ttttcgccgt aaggtgataa atcgccatgc tgcctcgctg ttgcttttgc
     5581 ttttcggctc catgcaatgg ccctcggaga gcgcaccgcc cgaagggtgg ccgttaggcc
     5641 agtttctcga agagaaaccg gtaagtgcgc cctcccctac aaagtagggt cgggattgcc
     5701 gccgctgtgc ctccatgata gcctacgaga cagcacatta acaatggggt gtcaagatgg
     5761 ttaaggggag caacaaggcg gcggatcggc tggccaagct cgaagaacaa cgagcgcgaa
     5821 tcaatgccga aattcagcgg gtgcgggcaa gggaacagca gcaagagcgc aagaacgaaa
     5881 caaggcgcaa ggtgctggtg ggggccatga ttttggccaa ggtgaacagc agcgagtggc
     5941 cggaggatcg gctcatggcg gcaatggatg cgtaccttga acgcgaccac gaccgcgcct
     6001 tgttcggtct gccgccacgc cagaaggatg agccgggctg aatgatcgac cgagacaggc
     6061 cctgcggggc tgcacacgcg cccccaccct tcgggtaggg ggaaaggccg ctaaagcggc
     6121 taaaagcgct ccagcgtatt tctgcggggt ttggtgtggg gtttagcggg ctttgcccgc
     6181 ctttccccct gccgcgcagc ggtggggcgg tgtgtagcct agcgcagcga atagaccagc
     6241 tatccggcct ctggccgggc atattgggca agggcagcag cgccccacaa gggcgctgat
     6301 aaccgcgcct agtggattat tcttagataa tcatggatgg atttttccaa caccccgcca
     6361 gcccccgccc ctgctgggtt tgcaggtttg ggggcgtgac agttattgca ggggttcgtg
     6421 acagttattg caggggggcg tgacagttat tgcaggggtt cgtgacagtt agtacgggag
     6481 tgacgggcac tggctggcaa tgtctagcaa cggcaggcat ttcggctgag ggtaaaagaa
     6541 ctttccgcta agcgatagac tgtatgtaaa cacagtattg caaggacgcg gaacatgcct
     6601 catgtggcgg ccaggacggc cagccgggat cgggatactg gtcgttacca gagccaccga
     6661 cccgagcaaa cccttctcta tcagatcgtt gacgagtatt acccggcatt cgctgcgctt
     6721 atggcagagc agggaaagga attgccgggc tatgtgcaac gggaatttga agaatttctc
     6781 caatgcgggc ggctggagca tggctttcta cgggttcgct gcgagtcttg ccacgccgag
     6841 cacctggtcg ctttcagaaa tcaatctaaa gtatatatga gtaaacttgg tctgacaggc
     6901 cccttgagac cagtccctat cagtgataga gattgacatc cctatcagtg atagagatac
     6961 tgagcacgga tctgaaagag gagaaaggat ctatggcgag tagcgaagac gttatcaaag
     7021 agttcatgcg tttcaaagtt cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg
     7081 aaggtgaagg tgaaggtcgt ccgtacgaag gtactcagac cgctaaactg aaagttacca
     7141 aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc gcagttccag tacggttcca
     7201 aagcttacgt taaacacccg gctgacatcc cggactacct gaaactgtcc ttcccggaag
     7261 gtttcaaatg ggaacgtgtt atgaacttcg aagacggtgg tgttgttacc gttacccagg
     7321 actcctccct gcaagacggt gagttcatct acaaagttaa actgcgtggt actaacttcc
     7381 cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta
     7441 tgtacccgga agacggtgct ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg
     7501 gtggtcacta cgacgctgaa gttaaaacca cctacatggc taaaaaaccg gttcagctgc
     7561 cgggtgctta caaaaccgac atcaaactgg acatcacctc ccacaacgaa gactacacca
     7621 tcgttgaaca gtacgaacgt gctgaaggtc gtcactccac cggtgcttaa taaggatctc
     7681 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     7741 tttgtcggtg aacgctctct ac
//
