LOCUS       sequence-374993-        7531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses red chromoprotein on a broad host range RSF1010 origin 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7531)
  AUTHORS   Elston KM, Phillips LE, Leonard SP, Young E, Holley JC, Ahsanullah 
            T, McReynolds B, Moran NA, Barrick JE
  TITLE     The Pathfinder plasmid toolkit for genetically engineering newly 
            isolated bacteria enables the study of Drosophila-colonizing 
            Orbaceae.
  JOURNAL   ISME Commun. 2023 May 24;3(1):49. doi: 10.1038/s43705-023-00255-3.
  PUBMED    37225918
REFERENCE   2  (bases 1 to 7531)
  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"; journalName: "ISME 
            Commun."; date: "2023-05-24"; volume: "3(1):49. doi"; pages: " 
            10.1038/s43705-023-00255-3"
COMMENT     Sequence Label: pBTK1004 (pSL3)
FEATURES             Location/Qualifiers
     source          1..7531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      537..931
                     /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)"
     oriT            1273..1360
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2599..3570
                     /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"
     CDS             4081..4920
                     /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             4907..5758
                     /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"
     promoter        join(5801..5806,5807..5823,5824..5829)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6018..6551
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     terminator      complement(6720..6747)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(6763..6834)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6854..7531)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 atgtaatatc tcctctgttt ctgtataaca gtactatgtg attataccag ccccctcact
       61 acatgtcaag aataaactgc caaagctggt ctcaaggggc ctgtcagacc aagtttactc
      121 atatatactt tagattgatt tctgaaagcg accaggtgct cggcgtggca agactcgcag
      181 cgaacccgta gaaagccatg ctccagccgc ccgcattgga gaaattcttc aaattcccgt
      241 tgcacatagc ccggcaattc ctttccctgc tctgccataa gcgcagcgaa tgccgggtaa
      301 tactcgtcaa cgatctgata gagaagggtt tgctcgggtc ggtggctctg gtaacgacca
      361 gtatcccgat cccggctggc cgtcctggcc gccacatgag gcatgttccg cgtccttgca
      421 atactgtgtt tacatacagt ctatcgctta gcggaaagtt cttttaccct cagccgaaat
      481 gcctgccgtt gctagacatt gccagccagt gcccgtcact cccgtactaa ctgtcacgaa
      541 cccctgcaat aactgtcacg cccccctgca ataactgtca cgaacccctg caataactgt
      601 cacgccccca aacctgcaaa cccagcaggg gcgggggctg gcggggtgtt ggaaaaatcc
      661 atccatgatt atctaagaat aatccactag gcgcggttat cagcgccctt gtggggcgct
      721 gctgcccttg cccaatatgc ccggccagag gccggatagc tggtctattc gctgcgctag
      781 gctacacacc gccccaccgc tgcgcggcag ggggaaaggc gggcaaagcc cgctaaaccc
      841 cacaccaaac cccgcagaaa tacgctggag cgcttttagc cgctttagcg gcctttcccc
      901 ctacccgaag ggtgggggcg cgtgtgcagc cccgcagggc ctgtctcggt cgatcattca
      961 gcccggctca tccttctggc gtggcggcag accgaacaag gcgcggtcgt ggtcgcgttc
     1021 aaggtacgca tccattgccg ccatgagccg atcctccggc cactcgctgc tgttcacctt
     1081 ggccaaaatc atggccccca ccagcacctt gcgccttgtt tcgttcttgc gctcttgctg
     1141 ctgttccctt gcccgcaccc gctgaatttc ggcattgatt cgcgctcgtt gttcttcgag
     1201 cttggccagc cgatccgccg ccttgttgct ccccttaacc atcttgacac cccattgtta
     1261 atgtgctgtc tcgtaggcta tcatggaggc acagcggcgg caatcccgac cctactttgt
     1321 aggggagggc gcacttaccg gtttctcttc gagaaactgg cctaacggcc acccttcggg
     1381 cggtgcgctc tccgagggcc attgcatgga gccgaaaagc aaaagcaaca gcgaggcagc
     1441 atggcgattt atcaccttac ggcgaaaacc ggcagcaggt cgggcggcca atcggccagg
     1501 gccaaggccg actacatcca gcgcgaaggc aagtatgccc gcgacatgga tgaagtcttg
     1561 cacgccgaat ccgggcacat gccggagttc gtcgagcggc ccgccgacta ctgggatgct
     1621 gccgacctgt atgaacgcgc caatgggcgg ctgttcaagg aggtcgaatt tgccctgccg
     1681 gtcgagctga ccctcgacca gcagaaggcg ctggcgtccg agttcgccca gcacctgacc
     1741 ggtgccgagc gcctgccgta tacgctggcc atccatgccg gtggcggcga gaacccgcac
     1801 tgccacctga tgatctccga gcggatcaat gacggcatcg agcggcccgc cgctcagtgg
     1861 ttcaagcggt acaacggcaa gaccccggag aagggcgggg cacagaagac cgaagcgctc
     1921 aagcccaagg catggcttga gcagacccgc gaggcatggg ccgaccatgc caaccgggca
     1981 ttagagcggg ctggccacga cgcccgcatt gaccacagaa cacttgaggc gcagggcatc
     2041 gagcgcctgc ccggtgttca cctggggccg aacgtggtgg agatggaagg ccggggcatc
     2101 cgcaccgacc gggcagacgt ggccctgaac atcgacaccg ccaacgccca gatcatcgac
     2161 ttacaggaat accgggaggc aatagaccat gaacgcaatc gacagagtga agaaatccag
     2221 aggcatcaac gagttagcgg agcagatcga accgctggcc cagagcatgg cgacactggc
     2281 cgacgaagcc cggcaggtca tgagccagac ccagcaggcc agcgaggcgc aggcggcgga
     2341 gtggctgaaa gcccagcgcc agacaggggc ggcatgggtg gagctggcca aagagttgcg
     2401 ggaggtagcc gccgaggtga gcagcgccgc gcagagcgcc cggagcgcgt cgcgggggtg
     2461 gcactggaag ctatggctaa ccgtgatgct ggcttccatg atgcctacgg tggtgctgct
     2521 gatcgcatcg ttgctcttgc tcgacctgac gccactgaca accgaggacg gctcgatctg
     2581 gctgcgcttg gtggcccgat gaagaacgac aggactttgc aggccatagg ccgacagctc
     2641 aaggccatgg gctgtgagcg cttcgatatc ggcgtcaggg acgccaccac cggccagatg
     2701 atgaaccggg aatggtcagc cgccgaagtg ctccagaaca cgccatggct caagcggatg
     2761 aatgcccagg gcaatgacgt gtatatcagg cccgccgagc aggagcggca tggtctggtg
     2821 ctggtggacg acctcagcga gtttgacctg gatgacatga aagccgaggg ccgggagcct
     2881 gccctggtag tggaaaccag cccgaagaac tatcaggcat gggtcaaggt ggccgacgcc
     2941 gcaggcggtg aacttcgggg gcagattgcc cggacgctgg ccagcgagta cgacgccgac
     3001 ccggccagcg ccgacagccg ccactatggc cgcttggcgg gcttcaccaa ccgcaaggac
     3061 aagcacacca cccgcgccgg ttatcagccg tgggtgctgc tgcgtgaatc caagggcaag
     3121 accgccaccg ctggcccggc gctggtgcag caggctggcc agcagatcga gcaggcccag
     3181 cggcagcagg agaaggcccg caggctggcc agcctcgaac tgcccgagcg gcagcttagc
     3241 cgccaccggc gcacggcgct ggacgagtac cgcagcgaga tggccgggct ggtcaagcgc
     3301 ttcggtgatg acctcagcaa gtgcgacttt atcgccgcgc agaagctggc cagccggggc
     3361 cgcagtgccg aggaaatcgg caaggccatg gccgaggcca gcccagcgct ggcagagcgc
     3421 aagcccggcc acgaagcgga ttacatcgag cgcaccgtca gcaaggtcat gggtctgccc
     3481 agcgtccagc ttgcgcgggc cgagctggca cgggcaccgg caccccgcca gcgaggcatg
     3541 gacaggggcg ggccagattt cagcatgtag tgcttgcgtt ggtactcacg cctgttatac
     3601 tatgagtact cacgcacaga agggggtttt atggaatacg aaaaaagcgc ttcagggtcg
     3661 gtctacctga tcaaaagtga caagggctat tggttgcccg gtggctttgg ttatacgtca
     3721 aacaaggccg aggctggccg cttttcagtc gctgatatgg ccagccttaa ccttgacggc
     3781 tgcaccttgt ccttgttccg cgaagacaag cctttcggcc ccggcaagtt tctcggtgac
     3841 tgatatgaaa gaccaaaagg acaagcagac cggcgacctg ctggccagcc ctgacgctgt
     3901 acgccaagcg cgatatgccg agcgcatgaa ggccaaaggg atgcgtcagc gcaagttctg
     3961 gctgaccgac gacgaatacg aggcgctgcg cgagtgcctg gaagaactca gagcggcgca
     4021 gggcgggggt agtgaccccg ccagcgccta accaccaact gcctgcaaag gaggcaatca
     4081 atggctaccc ataagcctat caatattctg gaggcgttcg cagcagcgcc gccaccgctg
     4141 gactacgttt tgcccaacat ggtggccggt acggtcgggg cgctggtgtc gcccggtggt
     4201 gccggtaaat ccatgctggc cctgcaactg gccgcacaga ttgcaggcgg gccggatctg
     4261 ctggaggtgg gcgaactgcc caccggcccg gtgatctacc tgcccgccga agacccgccc
     4321 accgccattc atcaccgcct gcacgccctt ggggcgcacc tcagcgccga ggaacggcaa
     4381 gccgtggctg acggcctgct gatccagccg ctgatcggca gcctgcccaa catcatggcc
     4441 ccggagtggt tcgacggcct caagcgcgcc gccgagggcc gccgcctgat ggtgctggac
     4501 acgctgcgcc ggttccacat cgaggaagaa aacgccagcg gccccatggc ccaggtcatc
     4561 ggtcgcatgg aggccatcgc cgccgatacc gggtgctcta tcgtgttcct gcaccatgcc
     4621 agcaagggcg cggccatgat gggcgcaggc gaccagcagc aggccagccg gggcagctcg
     4681 gtactggtcg ataacatccg ctggcagtcc tacctgtcga gcatgaccag cgccgaggcc
     4741 gaggaatggg gtgtggacga cgaccagcgc cggttcttcg tccgcttcgg tgtgagcaag
     4801 gccaactatg gcgcaccgtt cgctgatcgg tggttcaggc ggcatgacgg cggggtgctc
     4861 aagcccgccg tgctggagag gcagcgcaag agcaaggggg tgccccgtgg tgaagcctaa
     4921 gaacaagcac agcctcagcc acgtccggca cgacccggcg cactgtctgg cccccggcct
     4981 gttccgtgcc ctcaagcggg gcgagcgcaa gcgcagcaag ctggacgtga cgtatgacta
     5041 cggcgacggc aagcggatcg agttcagcgg cccggagccg ctgggcgctg atgatctgcg
     5101 catcctgcaa gggctggtgg ccatggctgg gcctaatggc ctagtgcttg gcccggaacc
     5161 caagaccgaa ggcggacggc agctccggct gttcctggaa cccaagtggg aggccgtcac
     5221 cgctgatgcc atggtggtca aaggtagcta tcgggcgctg gcaaaggaaa tcggggcaga
     5281 ggtcgatagt ggtggggcgc tcaagcacat acaggactgc atcgagcgcc tttggaaggt
     5341 atccatcatc gcccagaatg gccgcaagcg gcaggggttt cggctgctgt cggagtacgc
     5401 cagcgacgag gcggacgggc gcctgtacgt ggccctgaac cccttgatcg cgcaggccgt
     5461 catgggtggc ggccagcatg tgcgcatcag catggacgag gtgcgggcgc tggacagcga
     5521 aaccgcccgc ctgctgcacc agcggctgtg tggctggatc gaccccggca aaaccggcaa
     5581 ggcttccata gataccttgt gcggctatgt ctggccgtca gaggccagtg gttcgaccat
     5641 gcgcaagcgc cgccagcggg tgcgcgaggc gttgccggag ctggtcgcgc tgggctggac
     5701 ggtaaccgag ttcgcggcgg gcaagtacga catcacccgg cccaaggcgg caggctgact
     5761 cacgttaagg gattttggtc atgaacagca atggatcgaa ttgacataag cctgttcggt
     5821 tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     5881 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt
     5941 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     6001 gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc
     6061 ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg
     6121 accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag
     6181 ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga
     6241 cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg
     6301 ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg
     6361 gcgagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca
     6421 acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg
     6481 ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta
     6541 ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta
     6601 aattgtcaca acgccgcggc caattcgatc tagaattcca ttgagtaagt ttttaagcac
     6661 atcagcttca aaagcgctct gaagttccta gcggccgctc ggtgagaccg acttataaac
     6721 gcagaaaggc ccacccgaag gtgagccagt gtgactctag tagagagcgt tcaccgacaa
     6781 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     6841 ggagatcctt atattaagcg ccggtggagt gacgaccttc agcacgttcg tactgttcaa
     6901 cgatggtgta gtcttcgttg tgggaggtga tgtccagttt gatgtcggtt ttgtaagcac
     6961 ccggcagctg aaccggtttt ttagccatgt aggtggtttt aacttcagcg tcgtagtgac
     7021 caccgtcttt cagtttcaga cgcattttga tttcaccttt cagagcaccg tcttccgggt
     7081 acatacgttc ggtggaagct tcccaaccca tggttttttt ctgcataacc ggaccgtcgg
     7141 acgggaagtt ggtaccacgc agtttaactt tgtagatgaa ctcaccgtct tgcagggagg
     7201 agtcctgggt aacggtaaca acaccaccgt cttcgaagtt cataacacgt tcccatttga
     7261 aaccttccgg gaaggacagt ttcaggtagt ccgggatgtc agccgggtgt ttaacgtaag
     7321 ctttggaacc gtactggaac tgcggggaca ggatgtccca agcgaacggc agcggaccac
     7381 ctttggtaac tttcagttta gcggtctggg taccttcgta cggacgacct tcaccttcac
     7441 cttcgatttc gaactcgtga ccgttaacgg aaccttccat acgaactttg aaacgcatga
     7501 actctttgat aacgtcttcg gaggaagcca t
//
