LOCUS       sequence-208056-        7703 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 7703)
  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 7703)
  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: pBTK401
FEATURES             Location/Qualifiers
     source          1..7703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     621..640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(859..963)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1016..1867)
                     /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(1854..2693)
                     /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(3204..4175)
                     /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            5414..5501
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      5843..6237
                     /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    6689..6707
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6714..6732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             6750..6761
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6768..7445
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      7463..7534
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7550..7577
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(7701..7703,1..789,790..858))
                     /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"
ORIGIN
        1 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
       61 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      121 tgctgcaatg ataccgcggg acccacgctc accggctcca gatttatcag caataaacca
      181 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      241 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      301 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      361 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      421 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      481 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      541 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      601 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      661 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      721 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      781 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      841 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      901 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      961 gcgcacattt ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagtcagc
     1021 ctgccgcctt gggccgggtg atgtcgtact tgcccgccgc gaactcggtt accgtccagc
     1081 ccagcgcgac cagctccggc aacgcctcgc gcacccgctt gcggcgcttg cgcatggtcg
     1141 aaccactggc ctctgacggc cagacatagc cgcacaaggt atctatggaa gccttgccgg
     1201 ttttgccggg gtcgatccag ccacacagcc gctggtgcag caggcgggcg gtttcgctgt
     1261 ccagcgcccg cacctcgtcc atgctgatgc gcacatgctg gccgccaccc atgacggcct
     1321 gcgcgatcaa ggggttcagg gccacgtaca ggcgcccgtc cgcctcgtcg ctggcgtact
     1381 ccgacagcag ccgaaacccc tgccgcttgc ggccattctg ggcgatgatg gataccttcc
     1441 aaaggcgctc gatgcagtcc tgtatgtgct tgagcgcccc accactatcg acctctgccc
     1501 cgatttcctt tgccagcgcc cgatagctac ctttgaccac atggcattca gcggtgacgg
     1561 cctcccactt gggttccagg aacagccgga gctgccgtcc gccttcggtc ttgggttccg
     1621 ggccaagcac taggccatta ggcccagcca tggccaccag cccttgcagg atgcgcagat
     1681 catcagcgcc cagcggctcc gggccgctga actcgatccg cttgccgtcg ccgtagtcat
     1741 acgtcacgtc cagcttgctg cgcttgcgct cgccccgctt gagggcacgg aacaggccgg
     1801 gggccagaca gtgcgccggg tcgtgccgga cgtggctgag gctgtgcttg ttcttaggct
     1861 tcaccacggg gcaccccctt gctcttgcgc tgcctctcca gcacggcggg cttgagcacc
     1921 ccgccgtcat gccgcctgaa ccaccgatca gcgaacggtg cgccatagtt ggccttgctc
     1981 acaccgaagc ggacgaagaa ccggcgctgg tcgtcgtcca caccccattc ctcggcctcg
     2041 gcgctggtca tgctcgacag gtaggactgc cagcggatgt tatcgaccag taccgagctg
     2101 ccccggctgg cctgctgctg gtcgcctgcg cccatcatgg ccgcgccctt gctggcatgg
     2161 tgcaggaaca cgatagagca cccggtatcg gcggcgatgg cctccatgcg accgatgacc
     2221 tgggccatgg ggccgctggc gttttcttcc tcgatgtgga accggcgcag cgtgtccagc
     2281 accatcaggc ggcggccctc ggcggcgcgc ttgaggccgt cgaaccactc cggggccatg
     2341 atgttgggca ggctgccgat cagcggctgg atcagcaggc cgtcagccac ggcttgccgt
     2401 tcctcggcgc tgaggtgcgc cccaagggcg tgcaggcggt gatgaatggc ggtgggcggg
     2461 tcttcggcgg gcaggtagat caccgggccg gtgggcagtt cgcccacctc cagcagatcc
     2521 ggcccgcctg caatctgtgc ggccagttgc agggccagca tggatttacc ggcaccaccg
     2581 ggcgacacca gcgccccgac cgtaccggcc accatgttgg gcaaaacgta gtccagcggt
     2641 ggcggcgctg ctgcgaacgc ctccagaata ttgataggct tatgggtagc cattgattgc
     2701 ctcctttgca ggcagttggt ggttaggcgc tggcggggtc actacccccg ccctgcgccg
     2761 ctctgagttc ttccaggcac tcgcgcagcg cctcgtattc gtcgtcggtc agccagaact
     2821 tgcgctgacg catccctttg gccttcatgc gctcggcata tcgcgcttgg cgtacagcgt
     2881 cagggctggc cagcaggtcg ccggtctgct tgtccttttg gtctttcata tcagtcaccg
     2941 agaaacttgc cggggccgaa aggcttgtct tcgcggaaca aggacaaggt gcagccgtca
     3001 aggttaaggc tggccatatc agcgactgaa aagcggccag cctcggcctt gtttgacgta
     3061 taaccaaagc caccgggcaa ccaatagccc ttgtcacttt tgatcaggta gaccgaccct
     3121 gaagcgcttt tttcgtattc cataaaaccc ccttctgtgc gtgagtactc atagtataac
     3181 aggcgtgagt accaacgcaa gcactacatg ctgaaatctg gcccgcccct gtccatgcct
     3241 cgctggcggg gtgccggtgc ccgtgccagc tcggcccgcg caagctggac gctgggcaga
     3301 cccatgacct tgctgacggt gcgctcgatg taatccgctt cgtggccggg cttgcgctct
     3361 gccagcgctg ggctggcctc ggccatggcc ttgccgattt cctcggcact gcggccccgg
     3421 ctggccagct tctgcgcggc gataaagtcg cacttgctga ggtcatgacc gaagcgcttg
     3481 accagcccgg ccatctcgct gcggtactcg tccagcgccg tgcgccggtg gcggctaagc
     3541 tgccgctcgg gcagttcgag gctggccagc ctgcgggcct tctcctgctg ccgctgggcc
     3601 tgctcgatct gctggccagc ctgctgcacc agcgccgggc cagcggtggc ggtcttgccc
     3661 ttggattcac gcagcagcac ccacggctga taaccggcgc gggtggtgtg cttgtccttg
     3721 cggttggtga agcccgccaa gcggccatag tggcggctgt cggcgctggc cgggtcggcg
     3781 tcgtactcgc tggccagcgt ccgggcaatc tgcccccgaa gttcaccgcc tgcggcgtcg
     3841 gccaccttga cccatgcctg atagttcttc gggctggttt ccactaccag ggcaggctcc
     3901 cggccctcgg ctttcatgtc atccaggtca aactcgctga ggtcgtccac cagcaccaga
     3961 ccatgccgct cctgctcggc gggcctgata tacacgtcat tgccctgggc attcatccgc
     4021 ttgagccatg gcgtgttctg gagcacttcg gcggctgacc attcccggtt catcatctgg
     4081 ccggtgggtg cgtccctgac gccgatatcg aagcgctcac agcccatggc cttgagctgt
     4141 cggcctatgg cctgcaaagt cctgtcgttc ttcatcgggc caccaagcgc agccagatcg
     4201 agccgtcctc ggttgtcagt ggcgtcaggt cgagcaagag caacgatgcg atcagcagca
     4261 ccaccgtagg catcatggaa gccagcatca cggttagcca tagcttccag tgccaccccc
     4321 gcgacgcgct ccgggcgctc tgcgcggcgc tgctcacctc ggcggctacc tcccgcaact
     4381 ctttggccag ctccacccat gccgcccctg tctggcgctg ggctttcagc cactccgccg
     4441 cctgcgcctc gctggcctgc ttggtctggc tcatgacctg ccgggcttcg tcggccagtg
     4501 tcgccatgct ctgggccagc ggttcgatct gctccgctaa ctcgttgatg cctctggatt
     4561 tcttcactct gtcgattgcg ttcatggtct attgcctccc ggtattcctg taagtcgatg
     4621 atctgggcgt tggcggtgtc gatgttcagg gccacgtctg cccggtcggt gcggatgccc
     4681 cggccttcca tctccaccac gttcggcccc aggtgaacac cgggcaggcg ctcgatgccc
     4741 tgcgcctcaa gtgttctgtg gtcaatgcgg gcgtcgtggc cagcccgctc taatgcccgg
     4801 ttggcatggt cggcccatgc ctcgcgggtc tgctcaagcc atgccttggg cttgagcgct
     4861 tcggtcttct gtgccccgcc cttctccggg gtcttgccgt tgtaccgctt gaaccactga
     4921 gcggcgggcc gctcgatgcc gtcattgatc cgctcggaga tcatcaggtg gcagtgcggg
     4981 ttctcgccgc caccggcatg gatggccagc gtatacggca ggcgctcggc accggtcagg
     5041 tgctgggcga actcggacgc cagcgccttc tgctggtcga gggtcagctc gaccggcagg
     5101 gcaaattcga cctccttgaa cagccgccca ttggcgcgtt catacaggtc ggcagcatcc
     5161 cagtagtcgg cgggccgctc gacgaactcc ggcatgtgcc cggattcggc gtgcaagact
     5221 tcatccatgt cgcgggcata cttgccttcg cgctggatgt agtcggcctt ggccctggcc
     5281 gattggccgc ccgacctgct gccggttttc gccgtaaggt gataaatcgc catgctgcct
     5341 cgctgttgct tttgcttttc ggctccatgc aatggccctc ggagagcgca ccgcccgaag
     5401 ggtggccgtt aggccagttt ctcgaagaga aaccggtaag tgcgccctcc cctacaaagt
     5461 agggtcggga ttgccgccgc tgtgcctcca tgatagccta cgagacagca cattaacaat
     5521 ggggtgtcaa gatggttaag gggagcaaca aggcggcgga tcggctggcc aagctcgaag
     5581 aacaacgagc gcgaatcaat gccgaaattc agcgggagcg ggcaagggaa cagcagcaag
     5641 agcgcaagaa cgaaacaagg cgcaaggtgc tggtgggggc catgattttg gccaaggtga
     5701 acagcagcga gtggccggag gatcggctca tggcggcaat ggatgcgtac cttgaacgcg
     5761 accacgaccg cgccttgttc ggtctgccgc cacgccagaa ggatgagccg ggctgaatga
     5821 tcgaccgaga caggccctgc ggggctgcac acgcgccccc acccttcggg tagggggaaa
     5881 ggccgctaaa gcggctaaaa gcgctccagc gtatttctgc ggggtttggt gtggggttta
     5941 gcgggctttg cccgcctttc cccctgccgc gcagcggtgg ggcggtgtgt agcctagcgc
     6001 agcgaataga ccagctatcc ggcctctggc cgggcatatt gggcaagggc agcagcgccc
     6061 cacaagggcg ctgataaccg cgcctagtgg attattctta gataatcatg gatggatttt
     6121 tccaacaccc cgccagcccc cgcccctgct gggtttgcag gtttgggggc gtgacagtta
     6181 ttgcaggggt tcgtgacagt tattgcaggg gggcgtgaca gttattgcag gggttcgtga
     6241 cagttagtac gggagtgacg ggcactggct ggcaatgtct agcaacggca ggcatttcgg
     6301 ctgagggtaa aagaactttc cgctaagcga tagactgtat gtaaacacag tattgcaagg
     6361 acgcggaaca tgcctcatgt ggcggccagg acggccagcc gggatcggga tactggtcgt
     6421 taccagagcc accgacccga gcaaaccctt ctctatcaga tcgttgacga gtattacccg
     6481 gcattcgctg cgcttatggc agagcaggga aaggaattgc cgggctatgt gcaacgggaa
     6541 tttgaagaat ttctccaatg cgggcggctg gagcatggct ttctacgggt tcgctgcgag
     6601 tcttgccacg ccgagcacct ggtcgctttc agaaatcaat ctaaagtata tatgagtaaa
     6661 cttggtctga caggcccctt gagaccagtc cctatcagtg atagagattg acatccctat
     6721 cagtgataga gatactgagc acggatctga aagaggagaa aggatctatg gcgagtagcg
     6781 aagacgttat caaagagttc atgcgtttca aagttcgtat ggaaggttcc gttaacggtc
     6841 acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtact cagaccgcta
     6901 aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt
     6961 tccagtacgg ttccaaagct tacgttaaac acccggctga catcccggac tacctgaaac
     7021 tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg
     7081 ttaccgttac ccaggactcc tccctgcaag acggtgagtt catctacaaa gttaaactgc
     7141 gtggtactaa cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag
     7201 cttccaccga acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc
     7261 tgaaactgaa agacggtggt cactacgacg ctgaagttaa aaccacctac atggctaaaa
     7321 aaccggttca gctgccgggt gcttacaaaa ccgacatcaa actggacatc acctcccaca
     7381 acgaagacta caccatcgtt gaacagtacg aacgtgctga aggtcgtcac tccaccggtg
     7441 cttaataagg atctccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     7501 tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc
     7561 gggtgggcct ttctgcgttt ataagtcggt ctcaccgagc ggccgcgatt atcaaaaagg
     7621 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7681 gagtaaactt ggtctgacag tta
//
