LOCUS       sequence-218141-        8790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK assembled broad-host-range plasmid encoding GFP driven by CP25 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8790)
  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 8790)
  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: pBTK503
FEATURES             Location/Qualifiers
     source          1..8790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1661..1707)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1716..2339)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     2247..2266
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2379..2398)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2380..2398)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(2509..3300,3301..3369))
                     /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"
     primer_bind     3132..3151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3370..3474)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(3527..4378)
                     /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(4365..5204)
                     /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(5715..6686)
                     /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            7925..8012
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      8354..8748
                     /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)"
ORIGIN
        1 tagcaacggc aggcatttcg gctgagggta aaagaacttt ccgctaagcg atagactgta
       61 tgtaaacaca gtattgcaag gacgcggaac atgcctcatg tggcggccag gacggccagc
      121 cgggatcggg atactggtcg ttaccagagc caccgacccg agcaaaccct tctctatcag
      181 atcgttgacg agtattaccc ggcattcgct gcgcttatgg cagagcaggg aaaggaattg
      241 ccgggctatg tgcaacggga atttgaagaa tttctccaat gcgggcggct ggagcatggc
      301 tttctacggg ttcgctgcga gtcttgccac gccgagcacc tggtcgcttt cagaaatcaa
      361 tctaaagtat atatgagtaa acttggtctg acaggcccct gaattcgcat ctagactgat
      421 gagacgtggt agagccacaa acagccggta caagcaacga tctccaggac catctgaatc
      481 atgcgcggat gacacgaact cacgacggcg atcacagaca ttaacccaca gtacagacac
      541 tgcgacaacg tggcaattcg tcgcaataca acgctttggc agtttattct tgacatgtag
      601 tgagggggct ggtataatca catagtactg ttatacagaa acagaggaga tattacatat
      661 gagtaaagga gaagagcttt tcacaggagt tgtcccaatc ctcgtggaat tagacggtga
      721 tgttaatggg cacaagttct ctgtcagtgg agagggtgaa ggcgacgcaa catatggcaa
      781 gctgaccctt aaatttattt gcaccacggg taaactacct gttccatggc caacactggt
      841 cactacgttc gggtatgggg ttcagtgctt tgcgcgctac ccagatcaca tgaaacagca
      901 cgactttttc aagagtgcaa tgcccgaagg ctatgtacag gagagaacca tcttttttaa
      961 ggatgacggc aactataaga cacgcgccga agtgaagttc gagggtgata cccttgttaa
     1021 tagaatcgag ttaaagggta ttgactttaa ggaagatgga aatattttag gccacaaact
     1081 ggaatataac tataactccc ataatgtgta cattatggcc gacaagcaaa agaacggtat
     1141 caaggttaac ttcaagatca gacacaacat tgaggatgga agcgttcaac tagccgacca
     1201 ttaccaacaa aacaccccaa ttggcgatgg gcctgtgctg ttaccagaca accattacct
     1261 gtccactcaa tctgcccttt cgaaagatcc caacgaaaag cgcgaccaca tggtccttct
     1321 tgagtttgtc acggctgctg ggattacaca cggcatggat gaactataca aataaatccg
     1381 taatcgttaa tccgcaaata acgtaaaaac ccgcttcggc gggttttttt atggggggag
     1441 tttagggaaa gagcatttgt cagctggaaa tctgctcgtc agtggtgctc acactgacga
     1501 atcatgtaca gatcataccg atgactgcct ggcgactcac aactaagcaa gacagccgga
     1561 accagcgccg gcgaacacca ctgcatatat ggcatatcac aacagtccac gtctcaagca
     1621 gttacagaga tgttacgaac cactagtgca ctgcagtaca caaaaaaccc ctcaagaccc
     1681 gtttagaggc cccaaggggt tatgctaatg ctgctttaag acccactttc acatttaagt
     1741 tgtttttcta atccgcatat gatcaattca aggccgaata agaaggctgg ctctgcacct
     1801 tggtgatcaa ataattcgat agcttgtcgt aataatggcg gcatactatc agtagtaggt
     1861 gtttcccttt cttctttagc gacttgatgc tcttgatctt ccaatacgca acctaaagta
     1921 aaatgcccca ctgcgctgag tgcatataat gcattctcta gtgaaaaacc ttgttggcat
     1981 aaaaaggcta attgattttc gagagtttca tactgttttt ctgtaggccg tgtacctaaa
     2041 tgtacttttg ctccatcgcg atgacttagt aaagcacatc taaaactttt agcgttatta
     2101 cgtaaaaaat cttgccagct ttccccttct aaagggcaaa agtgagtatg gtgcctatct
     2161 aacatctcaa tggctaaggc gtcgagcaaa gcccgcttat tttttacatg ccaatacaat
     2221 gtaggctgct ctacacctag cttctgggcg agtttacggg ttgttaaacc ttcgattccg
     2281 acctcattaa gcagctctaa tgcgctgtta atcactttac ttttatctaa acgagacatg
     2341 agtatatctc cttctacaaa attatttcta gatctctccc ctatagtgag tcgtattaat
     2401 ttccgagcgg ccgcgattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2461 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2521 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2581 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2641 gataccgcgg gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2701 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2761 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2821 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2881 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2941 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3001 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3061 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3121 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3181 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3241 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3301 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3361 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3421 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3481 tccccgaaaa gtgccacctg tcatgaccaa aatcccttaa cgtgagtcag cctgccgcct
     3541 tgggccgggt gatgtcgtac ttgcccgccg cgaactcggt taccgtccag cccagcgcga
     3601 ccagctccgg caacgcctcg cgcacccgct ggcggcgctt gcgcatggtc gaaccactgg
     3661 cctctgacgg ccagacatag ccgcacaagg tatctatgga agccttgccg gttttgccgg
     3721 ggtcgatcca gccacacagc cgctggtgca gcaggcgggc ggtttcgctg tccagcgccc
     3781 gcacctcgtc catgctgatg cgcacatgct ggccgccacc catgacggcc tgcgcgatca
     3841 aggggttcag ggccacgtac aggcgcccgt ccgcctcgtc gctggcgtac tccgacagca
     3901 gccgaaaccc ctgccgcttg cggccattct gggcgatgat ggataccttc caaaggcgct
     3961 cgatgcagtc ctgtatgtgc ttgagcgccc caccactatc gacctctgcc ccgatttcct
     4021 ttgccagcgc ccgatagcta cctttgacca ccatggcatc agcggtgacg gcctcccact
     4081 tgggttccag gaacagccgg agctgccgtc cgccttcggt cttgggttcc gggccaagca
     4141 ctaggccatt aggcccagcc atggccacca gcccttgcag gatgcgcaga tcatcagcgc
     4201 ccagcggctc cgggccgctg aactcgatcc gcttgccgtc gccgtagtca tacgtcacgt
     4261 ccagcttgct gcgcttgcgc tcgccccgct tgagggcacg gaacaggccg ggggccagac
     4321 agtgcgccgg gtcgtgccgg acgtggctga ggctgtgctt gttcttaggc ttcaccacgg
     4381 ggcaccccct tgctcttgcg ctgcctctcc agcacggcgg gcttgagcac cccgccgtca
     4441 tgccgcctga accaccgatc agcgaacggt gcgccatagt tggccttgct cacaccgaag
     4501 cggacgaaga accggcgctg gtcgtcgtcc acaccccatt cctcggcctc ggcgctggtc
     4561 atgctcgaca ggtaggactg ccagcggatg ttatcgacca gtaccgagct gccccggctg
     4621 gcctgctgct ggtcgcctgc gcccatcatg gccgcgccct tgctggcatg gtgcaggaac
     4681 acgatagagc acccggtatc ggcggcgatg gcctccatgc gaccgatgac ctgggccatg
     4741 gggccgctgg cgttttcttc ctcgatgtgg aaccggcgca gcgtgtccag caccatcagg
     4801 cggcggccct cggcggcgcg cttgaggccg tcgaaccact ccggggccat gatgttgggc
     4861 aggctgccga tcagcggctg gatcagcagg ccgtcagcca cggcttgccg ttcctcggcg
     4921 ctgaggtgcg ccccaagggc gtgcaggcgg tgatgaatgg cggtgggcgg gtcttcggcg
     4981 ggcaggtaga tcaccgggcc ggtgggcagt tcgcccacct ccagcagatc cggcccgcct
     5041 gcaatctgtg cggccagttg cagggccagc atggatttac cggcaccacc gggcgacacc
     5101 agcgccccga ccgtaccggc caccatgttg ggcaaaacgt agtccagcgg tggcggcgct
     5161 gctgcgaacg cctccagaat attgataggc ttatgggtag ccattgattg cctcctttgc
     5221 aggcagttgg tggttaggcg ctggcggggt cactaccccc gccctgcgcc gctctgagtt
     5281 cttccaggca ctcgcgcagc gcctcgtatt cgtcgtcggt cagccagaac ttgcgctgac
     5341 gcatcccttt ggccttcatg cgctcggcat atcgcgcttg gcgtacagcg tcagggctgg
     5401 ccagcaggtc gccggtctgc ttgtcctttt ggtctttcat atcagtcacc gagaaacttg
     5461 ccggggccga aaggcttgtc ttcgcggaac aaggacaagg tgcagccgtc aaggttaagg
     5521 ctggccatat cagcgactga aaagcggcca gcctcggcct tgtttgacgt ataaccaaag
     5581 ccaccgggca accaatagcc cttgtcactt ttgatcaggt agaccgaccc tgaagcgctt
     5641 ttttcgtatt ccataaaacc cccttctgtg cgtgagtact catagtataa caggcgtgag
     5701 taccaacgca agcactacat gctgaaatct ggcccgcccc tgtccatgcc tcgctggcgg
     5761 ggtgccggtg cccgtgccag ctcggcccgc gcaagctgga cgctgggcag acccatgacc
     5821 ttgctgacgg tgcgctcgat gtaatccgct tcgtggccgg gcttgcgctc tgccagcgct
     5881 gggctggcct cggccatggc cttgccgatt tcctcggcac tgcggccccg gctggccagc
     5941 ttctgcgcgg cgataaagtc gcacttgctg aggtcatcac cgaagcgctt gaccagcccg
     6001 gccatctcgc tgcggtactc gtccagcgcc gtgcgccggt ggcggctaag ctgccgctcg
     6061 ggcagttcga ggctggccag cctgcgggcc ttctcctgct gccgctgggc ctgctcgatc
     6121 tgctggccag cctgctgcac cagcgccggg ccagcggtgg cggtcttgcc cttggattca
     6181 cgcagcagca cccacggctg ataaccggcg cgggtggtgt gcttgtcctt gcggttggtg
     6241 aagcccgcca agcggccata gtggcggctg tcggcgctgg ccgggtcggc gtcgtactcg
     6301 ctggccagcg tccgggcaat ctgcccccga agttcaccgc ctgcggcgtc ggccaccttg
     6361 acccatgcct gatagttctt cgggctggtt tccactacca gggcaggctc ccggccctcg
     6421 gctttcatgt catccaggtc aaactcgctg aggtcgtcca ccagcaccag accatgccgc
     6481 tcctgctcgg cgggcctgat atacacgtca ttgccctggg cattcatccg cttgagccat
     6541 ggcgtgttct ggagcacttc ggcggctgac cattcccggt tcatcatctg gccggtggtg
     6601 gcgtccctga cgccgatatc gaagcgctca cagcccatgg ccttgagctg tcggcctatg
     6661 gcctgcaaag tcctgtcgtt cttcatcggg ccaccaagcg cagccagatc gagccgtcct
     6721 cggttgtcag tggcgtcagg tcgagcaaga gcaacgatgc gatcagcagc accaccgtag
     6781 gcatcatgga agccagcatc acggttagcc atagcttcca gtgccacccc cgcgacgcgc
     6841 tccgggcgct ctgcgcggcg ctgctcacct cggcggctac ctcccgcaac tctttggcca
     6901 gctccaccca tgccgcccct gtctggcgct gggctttcag ccactccgcc gcctgcgcct
     6961 cgctggcctg ctgggtctgg ctcatgacct gccgggcttc gtcggccagt gtcgccatgc
     7021 tctgggccag cggttcgatc tgctccgcta actcgttgat gcctctggat ttcttcactc
     7081 tgtcgattgc gttcatggtc tattgcctcc cggtattcct gtaagtcgat gatctgggcg
     7141 ttggcggtgt cgatgttcag ggccacgtct gcccggtcgg tgcggatgcc ccggccttcc
     7201 atctccacca cgttcggccc caggtgaaca ccgggcaggc gctcgatgcc ctgcgcctca
     7261 agtgttctgt ggtcaatgcg ggcgtcgtgg ccagcccgct ctaatgcccg gttggcatgg
     7321 tcggcccatg cctcgcgggt ctgctcaagc catgccttgg gcttgagcgc ttcggtcttc
     7381 tgtgccccgc ccttctccgg ggtcttgccg ttgtaccgct tgaaccactg agcggcgggc
     7441 cgctcgatgc cgtcattgat ccgctcggag atcatcaggt ggcagtgcgg gttctcgccg
     7501 ccaccggcat ggatggccag cgtatacggc aggcgctcgg caccggtcag gtgctgggcg
     7561 aactcggacg ccagcgcctt ctgctggtcg agggtcagct cgaccggcag ggcaaattcg
     7621 acctccttga acagccgccc attggcgcgt tcatacaggt cggcagcatc ccagtagtcg
     7681 gcgggccgct cgacgaactc cggcatgtgc ccggattcgg cgtgcaagac ttcatccatg
     7741 tcgcgggcat acttgccttc gcgctggatg tagtcggcct tggccctggc cgattggccg
     7801 cccgacctgc tgccggtttt cgccgtaagg tgataaatcg ccatgctgcc tcgctgttgc
     7861 ttttgctttt cggctccatg caatggccct cggagagcgc accgcccgaa gggtggccgt
     7921 taggccagtt tctcgaagag aaaccggtaa gtgcgccctc ccctacaaag tagggtcggg
     7981 attgccgccg ctgtgcctcc atgatagcct acgagacagc acattaacaa tggggtgtca
     8041 agatggttaa ggggagcaac aaggcggcgg atcggctggc caagctcgaa gaacaacgag
     8101 cgcgaatcaa tgccgaaatt cagcgggtgc gggcaaggga acagcagcaa gagcgcaaga
     8161 acgaaacaag gcgcaaggtg ctggtggggg ccatgatttt ggccaaggtg aacagcagcg
     8221 agtggccgga ggatcggctc atggcggcaa tggatgcgta ccttgaacgc gaccacgacc
     8281 gcgccttgtt cggtctgccg ccacgccaga aggatgagcc gggctgaatg atcgaccgag
     8341 acaggccctg cggggctgca cacgcgcccc cacccttcgg gtagggggaa aggccgctaa
     8401 agcggctaaa agcgctccag cgtatttctg cggggtttgg tgtggggttt agcgggcttt
     8461 gcccgccttt ccccctgccg cgcagcggtg gggcggtgtg tagcctagcg cagcgaatag
     8521 accagctatc cggcctctgg ccgggcatat tgggcaaggg cagcagcgcc ccacaagggc
     8581 gctgataacc gcgcctagtg gattattctt agataatcat ggatggattt ttccaacacc
     8641 ccgccagccc ccgcccctgc tgggtttgca ggtttggggg cgtgacagtt attgcagggg
     8701 ttcgtgacag ttattgcagg ggggcgtgac agttattgca ggggttcgtg acagttagta
     8761 cgggagtgac gggcactggc tggcaatgtc
//
