LOCUS       sequence-208057-        7608 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 7608)
  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 7608)
  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: pBTK402
FEATURES             Location/Qualifiers
     source          1..7608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..816)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1..816)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     724..743
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     808..827
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(991..1842)
                     /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(1829..2668)
                     /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(3179..4150)
                     /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            5389..5476
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      5818..6212
                     /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    6664..6682
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6689..6707
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             6725..6736
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6743..7420
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      7438..7509
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7525..7552
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
       61 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      121 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
      181 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      241 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
      301 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
      361 cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
      421 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
      481 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
      541 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
      601 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
      661 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
      721 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
      781 cctggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
      841 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
      901 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
      961 tcagtcatga ccaaaatccc ttaacgtgag tcagcctgcc gccttgggcc gggtgatgtc
     1021 gtacttgccc gccgcgaact cggttaccgt ccagcccagc gcgaccagct ccggcaacgc
     1081 ctcgcgcacc cgcttgcggc gcttgcgcat ggtcgaacca ctggcctctg acggccagac
     1141 atagccgcac aaggtatcta tggaagcctt gccggttttg ccggggtcga tccagccaca
     1201 cagccgctgg tgcagcaggc gggcggtttc gctgtccagc gcccgcacct cgtccatgct
     1261 gatgcgcaca tgctggccgc cacccatgac ggcctgcgcg atcaaggggt tcagggccac
     1321 gtacaggcgc ccgtccgcct cgtcgctggc gtactccgac agcagccgaa acccctgccg
     1381 cttgcggcca ttctgggcga tgatggatac cttccaaagg cgctcgatgc agtcctgtat
     1441 gtgcttgagc gccccaccac tatcgacctc tgccccgatt tcctttgcca gcgcccgata
     1501 gctacctttg accacatggc attcagcggt gacggcctcc cacttgggtt ccaggaacag
     1561 ccggagctgc cgtccgcctt cggtcttggg ttccgggcca agcactaggc cattaggccc
     1621 agccatggcc accagccctt gcaggatgcg cagatcatca gcgcccagcg gctccgggcc
     1681 gctgaactcg atccgcttgc cgtcgccgta gtcatacgtc acgtccagct tgctgcgctt
     1741 gcgctcgccc cgcttgaggg cacggaacag gccgggggcc agacagtgcg ccgggtcgtg
     1801 ccggacgtgg ctgaggctgt gcttgttctt aggcttcacc acggggcacc cccttgctct
     1861 tgcgctgcct ctccagcacg gcgggcttga gcaccccgcc gtcatgccgc ctgaaccacc
     1921 gatcagcgaa cggtgcgcca tagttggcct tgctcacacc gaagcggacg aagaaccggc
     1981 gctggtcgtc gtccacaccc cattcctcgg cctcggcgct ggtcatgctc gacaggtagg
     2041 actgccagcg gatgttatcg accagtaccg agctgccccg gctggcctgc tgctggtcgc
     2101 ctgcgcccat catggccgcg cccttgctgg catggtgcag gaacacgata gagcacccgg
     2161 tatcggcggc gatggcctcc atgcgaccga tgacctgggc catggggccg ctggcgtttt
     2221 cttcctcgat gtggaaccgg cgcagcgtgt ccagcaccat caggcggcgg ccctcggcgg
     2281 cgcgcttgag gccgtcgaac cactccgggg ccatgatgtt gggcaggctg ccgatcagcg
     2341 gctggatcag caggccgtca gccacggctt gccgttcctc ggcgctgagg tgcgccccaa
     2401 gggcgtgcag gcggtgatga atggcggtgg gcgggtcttc ggcgggcagg tagatcaccg
     2461 ggccggtggg cagttcgccc acctccagca gatccggccc gcctgcaatc tgtgcggcca
     2521 gttgcagggc cagcatggat ttaccggcac caccgggcga caccagcgcc ccgaccgtac
     2581 cggccaccat gttgggcaaa acgtagtcca gcggtggcgg cgctgctgcg aacgcctcca
     2641 gaatattgat aggcttatgg gtagccattg attgcctcct ttgcaggcag ttggtggtta
     2701 ggcgctggcg gggtcactac ccccgccctg cgccgctctg agttcttcca ggcactcgcg
     2761 cagcgcctcg tattcgtcgt cggtcagcca gaacttgcgc tgacgcatcc ctttggcctt
     2821 catgcgctcg gcatatcgcg cttggcgtac agcgtcaggg ctggccagca ggtcgccggt
     2881 ctgcttgtcc ttttggtctt tcatatcagt caccgagaaa cttgccgggg ccgaaaggct
     2941 tgtcttcgcg gaacaaggac aaggtgcagc cgtcaaggtt aaggctggcc atatcagcga
     3001 ctgaaaagcg gccagcctcg gccttgtttg acgtataacc aaagccaccg ggcaaccaat
     3061 agcccttgtc acttttgatc aggtagaccg accctgaagc gcttttttcg tattccataa
     3121 aacccccttc tgtgcgtgag tactcatagt ataacaggcg tgagtaccaa cgcaagcact
     3181 acatgctgaa atctggcccg cccctgtcca tgcctcgctg gcggggtgcc ggtgcccgtg
     3241 ccagctcggc ccgcgcaagc tggacgctgg gcagacccat gaccttgctg acggtgcgct
     3301 cgatgtaatc cgcttcgtgg ccgggcttgc gctctgccag cgctgggctg gcctcggcca
     3361 tggccttgcc gatttcctcg gcactgcggc cccggctggc cagcttctgc gcggcgataa
     3421 agtcgcactt gctgaggtca tgaccgaagc gcttgaccag cccggccatc tcgctgcggt
     3481 actcgtccag cgccgtgcgc cggtggcggc taagctgccg ctcgggcagt tcgaggctgg
     3541 ccagcctgcg ggccttctcc tgctgccgct gggcctgctc gatctgctgg ccagcctgct
     3601 gcaccagcgc cgggccagcg gtggcggtct tgcccttgga ttcacgcagc agcacccacg
     3661 gctgataacc ggcgcgggtg gtgtgcttgt ccttgcggtt ggtgaagccc gccaagcggc
     3721 catagtggcg gctgtcggcg ctggccgggt cggcgtcgta ctcgctggcc agcgtccggg
     3781 caatctgccc ccgaagttca ccgcctgcgg cgtcggccac cttgacccat gcctgatagt
     3841 tcttcgggct ggtttccact accagggcag gctcccggcc ctcggctttc atgtcatcca
     3901 ggtcaaactc gctgaggtcg tccaccagca ccagaccatg ccgctcctgc tcggcgggcc
     3961 tgatatacac gtcattgccc tgggcattca tccgcttgag ccatggcgtg ttctggagca
     4021 cttcggcggc tgaccattcc cggttcatca tctggccggt gggtgcgtcc ctgacgccga
     4081 tatcgaagcg ctcacagccc atggccttga gctgtcggcc tatggcctgc aaagtcctgt
     4141 cgttcttcat cgggccacca agcgcagcca gatcgagccg tcctcggttg tcagtggcgt
     4201 caggtcgagc aagagcaacg atgcgatcag cagcaccacc gtaggcatca tggaagccag
     4261 catcacggtt agccatagct tccagtgcca cccccgcgac gcgctccggg cgctctgcgc
     4321 ggcgctgctc acctcggcgg ctacctcccg caactctttg gccagctcca cccatgccgc
     4381 ccctgtctgg cgctgggctt tcagccactc cgccgcctgc gcctcgctgg cctgcttggt
     4441 ctggctcatg acctgccggg cttcgtcggc cagtgtcgcc atgctctggg ccagcggttc
     4501 gatctgctcc gctaactcgt tgatgcctct ggatttcttc actctgtcga ttgcgttcat
     4561 ggtctattgc ctcccggtat tcctgtaagt cgatgatctg ggcgttggcg gtgtcgatgt
     4621 tcagggccac gtctgcccgg tcggtgcgga tgccccggcc ttccatctcc accacgttcg
     4681 gccccaggtg aacaccgggc aggcgctcga tgccctgcgc ctcaagtgtt ctgtggtcaa
     4741 tgcgggcgtc gtggccagcc cgctctaatg cccggttggc atggtcggcc catgcctcgc
     4801 gggtctgctc aagccatgcc ttgggcttga gcgcttcggt cttctgtgcc ccgcccttct
     4861 ccggggtctt gccgttgtac cgcttgaacc actgagcggc gggccgctcg atgccgtcat
     4921 tgatccgctc ggagatcatc aggtggcagt gcgggttctc gccgccaccg gcatggatgg
     4981 ccagcgtata cggcaggcgc tcggcaccgg tcaggtgctg ggcgaactcg gacgccagcg
     5041 ccttctgctg gtcgagggtc agctcgaccg gcagggcaaa ttcgacctcc ttgaacagcc
     5101 gcccattggc gcgttcatac aggtcggcag catcccagta gtcggcgggc cgctcgacga
     5161 actccggcat gtgcccggat tcggcgtgca agacttcatc catgtcgcgg gcatacttgc
     5221 cttcgcgctg gatgtagtcg gccttggccc tggccgattg gccgcccgac ctgctgccgg
     5281 ttttcgccgt aaggtgataa atcgccatgc tgcctcgctg ttgcttttgc ttttcggctc
     5341 catgcaatgg ccctcggaga gcgcaccgcc cgaagggtgg ccgttaggcc agtttctcga
     5401 agagaaaccg gtaagtgcgc cctcccctac aaagtagggt cgggattgcc gccgctgtgc
     5461 ctccatgata gcctacgaga cagcacatta acaatggggt gtcaagatgg ttaaggggag
     5521 caacaaggcg gcggatcggc tggccaagct cgaagaacaa cgagcgcgaa tcaatgccga
     5581 aattcagcgg gagcgggcaa gggaacagca gcaagagcgc aagaacgaaa caaggcgcaa
     5641 ggtgctggtg ggggccatga ttttggccaa ggtgaacagc agcgagtggc cggaggatcg
     5701 gctcatggcg gcaatggatg cgtaccttga acgcgaccac gaccgcgcct tgttcggtct
     5761 gccgccacgc cagaaggatg agccgggctg aatgatcgac cgagacaggc cctgcggggc
     5821 tgcacacgcg cccccaccct tcgggtaggg ggaaaggccg ctaaagcggc taaaagcgct
     5881 ccagcgtatt tctgcggggt ttggtgtggg gtttagcggg ctttgcccgc ctttccccct
     5941 gccgcgcagc ggtggggcgg tgtgtagcct agcgcagcga atagaccagc tatccggcct
     6001 ctggccgggc atattgggca agggcagcag cgccccacaa gggcgctgat aaccgcgcct
     6061 agtggattat tcttagataa tcatggatgg atttttccaa caccccgcca gcccccgccc
     6121 ctgctgggtt tgcaggtttg ggggcgtgac agttattgca ggggttcgtg acagttattg
     6181 caggggggcg tgacagttat tgcaggggtt cgtgacagtt agtacgggag tgacgggcac
     6241 tggctggcaa tgtctagcaa cggcaggcat ttcggctgag ggtaaaagaa ctttccgcta
     6301 agcgatagac tgtatgtaaa cacagtattg caaggacgcg gaacatgcct catgtggcgg
     6361 ccaggacggc cagccgggat cgggatactg gtcgttacca gagccaccga cccgagcaaa
     6421 cccttctcta tcagatcgtt gacgagtatt acccggcatt cgctgcgctt atggcagagc
     6481 agggaaagga attgccgggc tatgtgcaac gggaatttga agaatttctc caatgcgggc
     6541 ggctggagca tggctttcta cgggttcgct gcgagtcttg ccacgccgag cacctggtcg
     6601 ctttcagaaa tcaatctaaa gtatatatga gtaaacttgg tctgacaggc cccttgagac
     6661 cagtccctat cagtgataga gattgacatc cctatcagtg atagagatac tgagcacgga
     6721 tctgaaagag gagaaaggat ctatggcgag tagcgaagac gttatcaaag agttcatgcg
     6781 tttcaaagtt cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg
     6841 tgaaggtcgt ccgtacgaag gtactcagac cgctaaactg aaagttacca aaggtggtcc
     6901 gctgccgttc gcttgggaca tcctgtcccc gcagttccag tacggttcca aagcttacgt
     6961 taaacacccg gctgacatcc cggactacct gaaactgtcc ttcccggaag gtttcaaatg
     7021 ggaacgtgtt atgaacttcg aagacggtgg tgttgttacc gttacccagg actcctccct
     7081 gcaagacggt gagttcatct acaaagttaa actgcgtggt actaacttcc cgtccgacgg
     7141 tccggttatg cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta tgtacccgga
     7201 agacggtgct ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta
     7261 cgacgctgaa gttaaaacca cctacatggc taaaaaaccg gttcagctgc cgggtgctta
     7321 caaaaccgac atcaaactgg acatcacctc ccacaacgaa gactacacca tcgttgaaca
     7381 gtacgaacgt gctgaaggtc gtcactccac cggtgcttaa taaggatctc caggcatcaa
     7441 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     7501 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttataag
     7561 tcggtctcac cgagcggccg cgtgttacaa ccaattaacc aattctga
//
