LOCUS       sequence-374996-        7516 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 7516)
  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 7516)
  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: pBTK1005 (pSL4)
FEATURES             Location/Qualifiers
     source          1..7516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      687..758
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      774..801
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(831..1490)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1405..1424
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(1622..2473)
                     /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(2460..3299)
                     /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(3810..4781)
                     /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            6020..6107
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      6449..6843
                     /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)"
     CDS             join(7506..7516,1..667)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 cgaagacgtt atcaaagagt tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg
       61 tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta cccagaccgc
      121 taaactgaaa gttaccaaag gtggtccgct gccgttcgct tgggacatcc tgtccccgca
      181 gttccagtac ggttccaaag cttacgttaa acacccggct gacatcccgg actacctgaa
      241 actgtccttc ccggaaggtt tcaaatggga acgtgttatg aacttcgaag acggtggtgt
      301 tgttaccgtt acccaggact cctccctgca agacggtgag ttcatctaca aagttaaact
      361 gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca tgggttggga
      421 agcttccacc gaacgtatgt acccggaaga cggtgctctg aaaggtgaaa tcaaaatgcg
      481 tctgaaactg aaagacggtg gtcactacga cgctgaagtt aaaaccacct acatggctaa
      541 aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc aaactggaca tcacctccca
      601 caacgaagac tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg
      661 cgcttaatat aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
      721 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
      781 cttcgggtgg gcctttctgc gtttataagt cggtctcacc gagcggccgc ttacgccccg
      841 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
      901 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
      961 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     1021 tcaaaactgg tgaaactcac ccagggattg gctgaaacga aaaacatatt ctcaataaac
     1081 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     1141 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     1201 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     1261 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     1321 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     1381 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     1441 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     1501 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     1561 tggtgaaagt tggaacctct tacgtgcccg atcaatcatg accaaaatcc cttaacgtga
     1621 gtcagcctgc cgccttgggc cgggtgatgt cgtacttgcc cgccgcgaac tcggttaccg
     1681 tccagcccag cgcgaccagc tccggcaacg cctcgcgcac ccgctggcgg cgcttgcgca
     1741 tggtcgaacc actggcctct gacggccaga catagccgca caaggtatct atggaagcct
     1801 tgccggtttt gccggggtcg atccagccac acagccgctg gtgcagcagg cgggcggttt
     1861 cgctgtccag cgcccgcacc tcgtccatgc tgatgcgcac atgctggccg ccacccatga
     1921 cggcctgcgc gatcaagggg ttcagggcca cgtacaggcg cccgtccgcc tcgtcgctgg
     1981 cgtactccga cagcagccga aacccctgcc gcttgcggcc attctgggcg atgatggata
     2041 ccttccaaag gcgctcgatg cagtcctgta tgtgcttgag cgccccacca ctatcgacct
     2101 ctgccccgat ttcctttgcc agcgcccgat agctaccttt gaccaccatg gcatcagcgg
     2161 tgacggcctc ccacttgggt tccaggaaca gccggagctg ccgtccgcct tcggtcttgg
     2221 gttccgggcc aagcactagg ccattaggcc cagccatggc caccagccct tgcaggatgc
     2281 gcagatcatc agcgcccagc ggctccgggc cgctgaactc gatccgcttg ccgtcgccgt
     2341 agtcatacgt cacgtccagc ttgctgcgct tgcgctcgcc ccgcttgagg gcacggaaca
     2401 ggccgggggc cagacagtgc gccgggtcgt gccggacgtg gctgaggctg tgcttgttct
     2461 taggcttcac cacggggcac ccccttgctc ttgcgctgcc tctccagcac ggcgggcttg
     2521 agcaccccgc cgtcatgccg cctgaaccac cgatcagcga acggtgcgcc atagttggcc
     2581 ttgctcacac cgaagcggac gaagaaccgg cgctggtcgt cgtccacacc ccattcctcg
     2641 gcctcggcgc tggtcatgct cgacaggtag gactgccagc ggatgttatc gaccagtacc
     2701 gagctgcccc ggctggcctg ctgctggtcg cctgcgccca tcatggccgc gcccttgctg
     2761 gcatggtgca ggaacacgat agagcacccg gtatcggcgg cgatggcctc catgcgaccg
     2821 atgacctggg ccatggggcc gctggcgttt tcttcctcga tgtggaaccg gcgcagcgtg
     2881 tccagcacca tcaggcggcg gccctcggcg gcgcgcttga ggccgtcgaa ccactccggg
     2941 gccatgatgt tgggcaggct gccgatcagc ggctggatca gcaggccgtc agccacggct
     3001 tgccgttcct cggcgctgag gtgcgcccca agggcgtgca ggcggtgatg aatggcggtg
     3061 ggcgggtctt cggcgggcag gtagatcacc gggccggtgg gcagttcgcc cacctccagc
     3121 agatccggcc cgcctgcaat ctgtgcggcc agttgcaggg ccagcatgga tttaccggca
     3181 ccaccgggcg acaccagcgc cccgaccgta ccggccacca tgttgggcaa aacgtagtcc
     3241 agcggtggcg gcgctgctgc gaacgcctcc agaatattga taggcttatg ggtagccatt
     3301 gattgcctcc tttgcaggca gttggtggtt aggcgctggc ggggtcacta cccccgccct
     3361 gcgccgctct gagttcttcc aggcactcgc gcagcgcctc gtattcgtcg tcggtcagcc
     3421 agaacttgcg ctgacgcatc cctttggcct tcatgcgctc ggcatatcgc gcttggcgta
     3481 cagcgtcagg gctggccagc aggtcgccgg tctgcttgtc cttttggtct ttcatatcag
     3541 tcaccgagaa acttgccggg gccgaaaggc ttgtcttcgc ggaacaagga caaggtgcag
     3601 ccgtcaaggt taaggctggc catatcagcg actgaaaagc ggccagcctc ggccttgttt
     3661 gacgtataac caaagccacc gggcaaccaa tagcccttgt cacttttgat caggtagacc
     3721 gaccctgaag cgcttttttc gtattccata aaaccccctt ctgtgcgtga gtactcatag
     3781 tataacaggc gtgagtacca acgcaagcac tacatgctga aatctggccc gcccctgtcc
     3841 atgcctcgct ggcggggtgc cggtgcccgt gccagctcgg cccgcgcaag ctggacgctg
     3901 ggcagaccca tgaccttgct gacggtgcgc tcgatgtaat ccgcttcgtg gccgggcttg
     3961 cgctctgcca gcgctgggct ggcctcggcc atggccttgc cgatttcctc ggcactgcgg
     4021 ccccggctgg ccagcttctg cgcggcgata aagtcgcact tgctgaggtc atcaccgaag
     4081 cgcttgacca gcccggccat ctcgctgcgg tactcgtcca gcgccgtgcg ccggtggcgg
     4141 ctaagctgcc gctcgggcag ttcgaggctg gccagcctgc gggccttctc ctgctgccgc
     4201 tgggcctgct cgatctgctg gccagcctgc tgcaccagcg ccgggccagc ggtggcggtc
     4261 ttgcccttgg attcacgcag cagcacccac ggctgataac cggcgcgggt ggtgtgcttg
     4321 tccttgcggt tggtgaagcc cgccaagcgg ccatagtggc ggctgtcggc gctggccggg
     4381 tcggcgtcgt actcgctggc cagcgtccgg gcaatctgcc cccgaagttc accgcctgcg
     4441 gcgtcggcca ccttgaccca tgcctgatag ttcttcgggc tggtttccac taccagggca
     4501 ggctcccggc cctcggcttt catgtcatcc aggtcaaact cgctgaggtc gtccaccagc
     4561 accagaccat gccgctcctg ctcggcgggc ctgatataca cgtcattgcc ctgggcattc
     4621 atccgcttga gccatggcgt gttctggagc acttcggcgg ctgaccattc ccggttcatc
     4681 atctggccgg tggtggcgtc cctgacgccg atatcgaagc gctcacagcc catggccttg
     4741 agctgtcggc ctatggcctg caaagtcctg tcgttcttca tcgggccacc aagcgcagcc
     4801 agatcgagcc gtcctcggtt gtcagtggcg tcaggtcgag caagagcaac gatgcgatca
     4861 gcagcaccac cgtaggcatc atggaagcca gcatcacggt tagccatagc ttccagtgcc
     4921 acccccgcga cgcgctccgg gcgctctgcg cggcgctgct cacctcggcg gctacctccc
     4981 gcaactcttt ggccagctcc acccatgccg cccctgtctg gcgctgggct ttcagccact
     5041 ccgccgcctg cgcctcgctg gcctgctggg tctggctcat gacctgccgg gcttcgtcgg
     5101 ccagtgtcgc catgctctgg gccagcggtt cgatctgctc cgctaactcg ttgatgcctc
     5161 tggatttctt cactctgtcg attgcgttca tggtctattg cctcccggta ttcctgtaag
     5221 tcgatgatct gggcgttggc ggtgtcgatg ttcagggcca cgtctgcccg gtcggtgcgg
     5281 atgccccggc cttccatctc caccacgttc ggccccaggt gaacaccggg caggcgctcg
     5341 atgccctgcg cctcaagtgt tctgtggtca atgcgggcgt cgtggccagc ccgctctaat
     5401 gcccggttgg catggtcggc ccatgcctcg cgggtctgct caagccatgc cttgggcttg
     5461 agcgcttcgg tcttctgtgc cccgcccttc tccggggtct tgccgttgta ccgcttgaac
     5521 cactgagcgg cgggccgctc gatgccgtca ttgatccgct cggagatcat caggtggcag
     5581 tgcgggttct cgccgccacc ggcatggatg gccagcgtat acggcaggcg ctcggcaccg
     5641 gtcaggtgct gggcgaactc ggacgccagc gccttctgct ggtcgagggt cagctcgacc
     5701 ggcagggcaa attcgacctc cttgaacagc cgcccattgg cgcgttcata caggtcggca
     5761 gcatcccagt agtcggcggg ccgctcgacg aactccggca tgtgcccgga ttcggcgtgc
     5821 aagacttcat ccatgtcgcg ggcatacttg ccttcgcgct ggatgtagtc ggccttggcc
     5881 ctggccgatt ggccgcccga cctgctgccg gttttcgccg taaggtgata aatcgccatg
     5941 ctgcctcgct gttgcttttg cttttcggct ccatgcaatg gccctcggag agcgcaccgc
     6001 ccgaagggtg gccgttaggc cagtttctcg aagagaaacc ggtaagtgcg ccctccccta
     6061 caaagtaggg tcgggattgc cgccgctgtg cctccatgat agcctacgag acagcacatt
     6121 aacaatgggg tgtcaagatg gttaagggga gcaacaaggc ggcggatcgg ctggccaagc
     6181 tcgaagaaca acgagcgcga atcaatgccg aaattcagcg ggtgcgggca agggaacagc
     6241 agcaagagcg caagaacgaa acaaggcgca aggtgctggt gggggccatg attttggcca
     6301 aggtgaacag cagcgagtgg ccggaggatc ggctcatggc ggcaatggat gcgtaccttg
     6361 aacgcgacca cgaccgcgcc ttgttcggtc tgccgccacg ccagaaggat gagccgggct
     6421 gaatgatcga ccgagacagg ccctgcgggg ctgcacacgc gcccccaccc ttcgggtagg
     6481 gggaaaggcc gctaaagcgg ctaaaagcgc tccagcgtat ttctgcgggg tttggtgtgg
     6541 ggtttagcgg gctttgcccg cctttccccc tgccgcgcag cggtggggcg gtgtgtagcc
     6601 tagcgcagcg aatagaccag ctatccggcc tctggccggg catattgggc aagggcagca
     6661 gcgccccaca agggcgctga taaccgcgcc tagtggatta ttcttagata atcatggatg
     6721 gatttttcca acaccccgcc agcccccgcc cctgctgggt ttgcaggttt gggggcgtga
     6781 cagttattgc aggggttcgt gacagttatt gcaggggggc gtgacagtta ttgcaggggt
     6841 tcgtgacagt tagtacggga gtgacgggca ctggctggca atgtctagca acggcaggca
     6901 tttcggctga gggtaaaaga actttccgct aagcgataga ctgtatgtaa acacagtatt
     6961 gcaaggacgc ggaacatgcc tcatgtggcg gccaggacgg ccagccggga tcgggatact
     7021 ggtcgttacc agagccaccg acccgagcaa acccttctct atcagatcgt tgacgagtat
     7081 tacccggcat tcgctgcgct tatggcagag cagggaaagg aattgccggg ctatgtgcaa
     7141 cgggaatttg aagaatttct ccaatgcggg cggctggagc atggctttct acgggttcgc
     7201 tgcgagtctt gccacgccga gcacctggtc gctttcagaa atcaatctaa agtatatatg
     7261 agtaaacttg gtctgacagg ccccttgaga ccagctttgg cagtttattc ttgacatgta
     7321 gtgagggggc tggtataatc acatagtact gttatacaga aacagaggag atattacata
     7381 tggcttcctc cgaagacgtt atcaggcccc ttgagaccag ctttggcagt ttattcttga
     7441 catgtagtga gggggctggt ataatcacat agtactgtta tacagaaaca gaggagatat
     7501 tacatatggc ttcctc
//
