LOCUS       sequence-374992-        7923 bp    DNA     circular SYN 14-APR-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 7923)
  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 7923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pBTK1003 (pSL2)
FEATURES             Location/Qualifiers
     source          1..7923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      674..1068
                     /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            1410..1497
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2736..3707
                     /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             4218..5057
                     /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             5044..5895
                     /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             6275..7066
                     /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"
     terminator      complement(7123..7150)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(7166..7237)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(7257..7923,1..11))
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 gaggaagcca tatgtaatat ctcctctgtt tctgtataac agtactatgt gattatacca
       61 gccccctcac tacatgtcaa gaataaactg ccaaagctgg tctcaagggg cctgataacg
      121 tcttcggagg aagccatatg taatatctcc tctgtttctg tataacagta ctatgtgatt
      181 ataccagccc cctcactaca tgtcaagaat aaactgccaa agctggtctc aaggggcctg
      241 tcagaccaag tttactcata tatactttag attgatttct gaaagcgacc aggtgctcgg
      301 cgtggcaaga ctcgcagcga acccgtagaa agccatgctc cagccgcccg cattggagaa
      361 attcttcaaa ttcccgttgc acatagcccg gcaattcctt tccctgctct gccataagcg
      421 cagcgaatgc cgggtaatac tcgtcaacga tctgatagag aagggtttgc tcgggtcggt
      481 ggctctggta acgaccagta tcccgatccc ggctggccgt cctggccgcc acatgaggca
      541 tgttccgcgt ccttgcaata ctgtgtttac atacagtcta tcgcttagcg gaaagttctt
      601 ttaccctcag ccgaaatgcc tgccgttgct agacattgcc agccagtgcc cgtcactccc
      661 gtactaactg tcacgaaccc ctgcaataac tgtcacgccc ccctgcaata actgtcacga
      721 acccctgcaa taactgtcac gcccccaaac ctgcaaaccc agcaggggcg ggggctggcg
      781 gggtgttgga aaaatccatc catgattatc taagaataat ccactaggcg cggttatcag
      841 cgcccttgtg gggcgctgct gcccttgccc aatatgcccg gccagaggcc ggatagctgg
      901 tctattcgct gcgctaggct acacaccgcc ccaccgctgc gcggcagggg gaaaggcggg
      961 caaagcccgc taaaccccac accaaacccc gcagaaatac gctggagcgc ttttagccgc
     1021 tttagcggcc tttcccccta cccgaagggt gggggcgcgt gtgcagcccc gcagggcctg
     1081 tctcggtcga tcattcagcc cggctcatcc ttctggcgtg gcggcagacc gaacaaggcg
     1141 cggtcgtggt cgcgttcaag gtacgcatcc attgccgcca tgagccgatc ctccggccac
     1201 tcgctgctgt tcaccttggc caaaatcatg gcccccacca gcaccttgcg ccttgtttcg
     1261 ttcttgcgct cttgctgctg ttcccttgcc cgcacccgct gaatttcggc attgattcgc
     1321 gctcgttgtt cttcgagctt ggccagccga tccgccgcct tgttgctccc cttaaccatc
     1381 ttgacacccc attgttaatg tgctgtctcg taggctatca tggaggcaca gcggcggcaa
     1441 tcccgaccct actttgtagg ggagggcgca cttaccggtt tctcttcgag aaactggcct
     1501 aacggccacc cttcgggcgg tgcgctctcc gagggccatt gcatggagcc gaaaagcaaa
     1561 agcaacagcg aggcagcatg gcgatttatc accttacggc gaaaaccggc agcaggtcgg
     1621 gcggccaatc ggccagggcc aaggccgact acatccagcg cgaaggcaag tatgcccgcg
     1681 acatggatga agtcttgcac gccgaatccg ggcacatgcc ggagttcgtc gagcggcccg
     1741 ccgactactg ggatgctgcc gacctgtatg aacgcgccaa tgggcggctg ttcaaggagg
     1801 tcgaatttgc cctgccggtc gagctgaccc tcgaccagca gaaggcgctg gcgtccgagt
     1861 tcgcccagca cctgaccggt gccgagcgcc tgccgtatac gctggccatc catgccggtg
     1921 gcggcgagaa cccgcactgc cacctgatga tctccgagcg gatcaatgac ggcatcgagc
     1981 ggcccgccgc tcagtggttc aagcggtaca acggcaagac cccggagaag ggcggggcac
     2041 agaagaccga agcgctcaag cccaaggcat ggcttgagca gacccgcgag gcatgggccg
     2101 accatgccaa ccgggcatta gagcgggctg gccacgacgc ccgcattgac cacagaacac
     2161 ttgaggcgca gggcatcgag cgcctgcccg gtgttcacct ggggccgaac gtggtggaga
     2221 tggaaggccg gggcatccgc accgaccggg cagacgtggc cctgaacatc gacaccgcca
     2281 acgcccagat catcgactta caggaatacc gggaggcaat agaccatgaa cgcaatcgac
     2341 agagtgaaga aatccagagg catcaacgag ttagcggagc agatcgaacc gctggcccag
     2401 agcatggcga cactggccga cgaagcccgg caggtcatga gccagaccca gcaggccagc
     2461 gaggcgcagg cggcggagtg gctgaaagcc cagcgccaga caggggcggc atgggtggag
     2521 ctggccaaag agttgcggga ggtagccgcc gaggtgagca gcgccgcgca gagcgcccgg
     2581 agcgcgtcgc gggggtggca ctggaagcta tggctaaccg tgatgctggc ttccatgatg
     2641 cctacggtgg tgctgctgat cgcatcgttg ctcttgctcg acctgacgcc actgacaacc
     2701 gaggacggct cgatctggct gcgcttggtg gcccgatgaa gaacgacagg actttgcagg
     2761 ccataggccg acagctcaag gccatgggct gtgagcgctt cgatatcggc gtcagggacg
     2821 ccaccaccgg ccagatgatg aaccgggaat ggtcagccgc cgaagtgctc cagaacacgc
     2881 catggctcaa gcggatgaat gcccagggca atgacgtgta tatcaggccc gccgagcagg
     2941 agcggcatgg tctggtgctg gtggacgacc tcagcgagtt tgacctggat gacatgaaag
     3001 ccgagggccg ggagcctgcc ctggtagtgg aaaccagccc gaagaactat caggcatggg
     3061 tcaaggtggc cgacgccgca ggcggtgaac ttcgggggca gattgcccgg acgctggcca
     3121 gcgagtacga cgccgacccg gccagcgccg acagccgcca ctatggccgc ttggcgggct
     3181 tcaccaaccg caaggacaag cacaccaccc gcgccggtta tcagccgtgg gtgctgctgc
     3241 gtgaatccaa gggcaagacc gccaccgctg gcccggcgct ggtgcagcag gctggccagc
     3301 agatcgagca ggcccagcgg cagcaggaga aggcccgcag gctggccagc ctcgaactgc
     3361 ccgagcggca gcttagccgc caccggcgca cggcgctgga cgagtaccgc agcgagatgg
     3421 ccgggctggt caagcgcttc ggtgatgacc tcagcaagtg cgactttatc gccgcgcaga
     3481 agctggccag ccggggccgc agtgccgagg aaatcggcaa ggccatggcc gaggccagcc
     3541 cagcgctggc agagcgcaag cccggccacg aagcggatta catcgagcgc accgtcagca
     3601 aggtcatggg tctgcccagc gtccagcttg cgcgggccga gctggcacgg gcaccggcac
     3661 cccgccagcg aggcatggac aggggcgggc cagatttcag catgtagtgc ttgcgttggt
     3721 actcacgcct gttatactat gagtactcac gcacagaagg gggttttatg gaatacgaaa
     3781 aaagcgcttc agggtcggtc tacctgatca aaagtgacaa gggctattgg ttgcccggtg
     3841 gctttggtta tacgtcaaac aaggccgagg ctggccgctt ttcagtcgct gatatggcca
     3901 gccttaacct tgacggctgc accttgtcct tgttccgcga agacaagcct ttcggccccg
     3961 gcaagtttct cggtgactga tatgaaagac caaaaggaca agcagaccgg cgacctgctg
     4021 gccagccctg acgctgtacg ccaagcgcga tatgccgagc gcatgaaggc caaagggatg
     4081 cgtcagcgca agttctggct gaccgacgac gaatacgagg cgctgcgcga gtgcctggaa
     4141 gaactcagag cggcgcaggg cgggggtagt gaccccgcca gcgcctaacc accaactgcc
     4201 tgcaaaggag gcaatcaatg gctacccata agcctatcaa tattctggag gcgttcgcag
     4261 cagcgccgcc accgctggac tacgttttgc ccaacatggt ggccggtacg gtcggggcgc
     4321 tggtgtcgcc cggtggtgcc ggtaaatcca tgctggccct gcaactggcc gcacagattg
     4381 caggcgggcc ggatctgctg gaggtgggcg aactgcccac cggcccggtg atctacctgc
     4441 ccgccgaaga cccgcccacc gccattcatc accgcctgca cgcccttggg gcgcacctca
     4501 gcgccgagga acggcaagcc gtggctgacg gcctgctgat ccagccgctg atcggcagcc
     4561 tgcccaacat catggccccg gagtggttcg acggcctcaa gcgcgccgcc gagggccgcc
     4621 gcctgatggt gctggacacg ctgcgccggt tccacatcga ggaagaaaac gccagcggcc
     4681 ccatggccca ggtcatcggt cgcatggagg ccatcgccgc cgataccggg tgctctatcg
     4741 tgttcctgca ccatgccagc aagggcgcgg ccatgatggg cgcaggcgac cagcagcagg
     4801 ccagccgggg cagctcggta ctggtcgata acatccgctg gcagtcctac ctgtcgagca
     4861 tgaccagcgc cgaggccgag gaatggggtg tggacgacga ccagcgccgg ttcttcgtcc
     4921 gcttcggtgt gagcaaggcc aactatggcg caccgttcgc tgatcggtgg ttcaggcggc
     4981 atgacggcgg ggtgctcaag cccgccgtgc tggagaggca gcgcaagagc aagggggtgc
     5041 cccgtggtga agcctaagaa caagcacagc ctcagccacg tccggcacga cccggcgcac
     5101 tgtctggccc ccggcctgtt ccgtgccctc aagcggggcg agcgcaagcg cagcaagctg
     5161 gacgtgacgt atgactacgg cgacggcaag cggatcgagt tcagcggccc ggagccgctg
     5221 ggcgctgatg atctgcgcat cctgcaaggg ctggtggcca tggctgggcc taatggccta
     5281 gtgcttggcc cggaacccaa gaccgaaggc ggacggcagc tccggctgtt cctggaaccc
     5341 aagtgggagg ccgtcaccgc tgatgccatg gtggtcaaag gtagctatcg ggcgctggca
     5401 aaggaaatcg gggcagaggt cgatagtggt ggggcgctca agcacataca ggactgcatc
     5461 gagcgccttt ggaaggtatc catcatcgcc cagaatggcc gcaagcggca ggggtttcgg
     5521 ctgctgtcgg agtacgccag cgacgaggcg gacgggcgcc tgtacgtggc cctgaacccc
     5581 ttgatcgcgc aggccgtcat gggtggcggc cagcatgtgc gcatcagcat ggacgaggtg
     5641 cgggcgctgg acagcgaaac cgcccgcctg ctgcaccagc ggctgtgtgg ctggatcgac
     5701 cccggcaaaa ccggcaaggc ttccatagat accttgtgcg gctatgtctg gccgtcagag
     5761 gccagtggtt cgaccatgcg caagcgccgc cagcgggtgc gcgaggcgtt gccggagctg
     5821 gtcgcgctgg gctggacggt aaccgagttc gcggcgggca agtacgacat cacccggccc
     5881 aaggcggcag gctgactcac gttaagggat tttggtcatg accagccagg acagaaatgc
     5941 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     6001 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     6061 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     6121 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     6181 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     6241 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     6301 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     6361 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     6421 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     6481 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     6541 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     6601 tggagaatgg cagcgcaatg acattcttgc gggtatcttc gagccagcca cgatcgacat
     6661 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     6721 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     6781 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     6841 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     6901 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     6961 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     7021 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataatcag aattggttaa
     7081 ttggttgtaa cacgcggccg ctcggtgaga ccgacttata aacgcagaaa ggcccacccg
     7141 aaggtgagcc agtgtgactc tagtagagag cgttcaccga caaacaacag ataaaacgaa
     7201 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctggagatc cttatattaa
     7261 gcgccggtgg agtgacgacc ttcagcacgt tcgtactgtt caacgatggt gtagtcttcg
     7321 ttgtgggagg tgatgtccag tttgatgtcg gttttgtaag cacccggcag ctgaaccggt
     7381 tttttagcca tgtaggtggt tttaacttca gcgtcgtagt gaccaccgtc tttcagtttc
     7441 agacgcattt tgatttcacc tttcagagca ccgtcttccg ggtacatacg ttcggtggaa
     7501 gcttcccaac ccatggtttt tttctgcata accggaccgt cggacgggaa gttggtacca
     7561 cgcagtttaa ctttgtagat gaactcaccg tcttgcaggg aggagtcctg ggtaacggta
     7621 acaacaccac cgtcttcgaa gttcataaca cgttcccatt tgaaaccttc cgggaaggac
     7681 agtttcaggt agtccgggat gtcagccggg tgtttaacgt aagctttgga accgtactgg
     7741 aactgcgggg acaggatgtc ccaagcgaac ggcagcggac cacctttggt aactttcagt
     7801 ttagcggtct gggtaccttc gtacggacga ccttcacctt caccttcgat ttcgaactcg
     7861 tgaccgttaa cggaaccttc catacgaact ttgaaacgca tgaactcttt gataacgtct
     7921 tcg
//
