LOCUS       sequence-315024-        8807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RSF1010 origin, LuxR activator, 
            PLuxB promoter, mRFP, Chloramphenicol Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8807)
  AUTHORS   Schuster LA, Reisch CR
  TITLE     A plasmid toolbox for controlled gene expression across the 
            Proteobacteria.
  JOURNAL   Nucleic Acids Res. 2021 Jun 14. pii: 6298615. doi: 
            10.1093/nar/gkab496.
  PUBMED    34125913
REFERENCE   2  (bases 1 to 8807)
  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: "Nucleic 
            Acids Res. 2021 Jun 14. pii: 6298615. doi: 10.1093/nar/gkab496."
COMMENT     Sequence Label: pFLxR1
FEATURES             Location/Qualifiers
     source          1..8807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      330..724
                     /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            1066..1153
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2392..3363
                     /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             3874..4713
                     /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             4700..5551
                     /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"
     primer_bind     5975..5997
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5989..6006
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5990..6006
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        6984..7033
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             7078..7089
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7096..7773
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     8441..8460
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(8527..8629)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(8746..8764)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtggcactt ttcggggatc ttcaaattcc cgttgcacat agcccggcaa ttcctttccc
       61 tgctctgcca taagcgcagc gaatgccggg taatactcgt caacgatctg atagagaagg
      121 gtttgctcgg gtcggtggct ctggtaacga ccagtatccc gatcccggct ggccgtcctg
      181 gccgccacat gaggcatgtt ccgcgtcctt gcaatactgt gtttacatac agtctatcgc
      241 ttagcggaaa gttcttttac cctcagccga aatgcctgcc gttgctagac attgccagcc
      301 agtgcccgtc actcccgtac taactgtcac gaacccctgc aataactgtc acgcccccct
      361 gcaataactg tcacgaaccc ctgcaataac tgtcacgccc ccaaacctgc aaacccagca
      421 ggggcggggg ctggcggggt gttggaaaaa tccatccatg attatctaag aataatccac
      481 taggcgcggt tatcagcgcc cttgtggggc gctgctgccc ttgcccaata tgcccggcca
      541 gaggccggat agctggtcta ttcgctgcgc taggctacac accgccccac cgctgcgcgg
      601 cagggggaaa ggcgggcaaa gcccgctaaa ccccacacca aaccccgcag aaatacgctg
      661 gagcgctttt agccgcttta gcggcctttc cccctacccg aagggtgggg gcgcgtgtgc
      721 agccccgcag ggcctgtctc ggtcgatcat tcagcccggc tcatccttct ggcgtggcgg
      781 cagaccgaac aaggcgcggt cgtggtcgcg ttcaaggtac gcatccattg ccgccatgag
      841 ccgatcctcc ggccactcgc tgctgttcac cttggccaaa atcatggccc ccaccagcac
      901 cttgcgcctt gtttcgttct tgcgctcttg ctgctgttcc cttgcccgca cccgctgaat
      961 ttcggcattg attcgcgctc gttgttcttc gagcttggcc agccgatccg ccgccttgtt
     1021 gctcccctta accatcttga caccccattg ttaatgtgct gtctcgtagg ctatcatgga
     1081 ggcacagcgg cggcaatccc gaccctactt tgtaggggag ggcgcactta ccggtttctc
     1141 ttcgagaaac tggcctaacg gccacccttc gggcggtgcg ctctccgagg gccattgcat
     1201 ggagccgaaa agcaaaagca acagcgaggc agcatggcga tttatcacct tacggcgaaa
     1261 accggcagca ggtcgggcgg ccaatcggcc agggccaagg ccgactacat ccagcgcgaa
     1321 ggcaagtatg cccgcgacat ggatgaagtc ttgcacgccg aatccgggca catgccggag
     1381 ttcgtcgagc ggcccgccga ctactgggat gctgccgacc tgtatgaacg cgccaatggg
     1441 cggctgttca aggaggtcga atttgccctg ccggtcgagc tgaccctcga ccagcagaag
     1501 gcgctggcgt ccgagttcgc ccagcacctg accggtgccg agcgcctgcc gtatacgctg
     1561 gccatccatg ccggtggcgg cgagaacccg cactgccacc tgatgatctc cgagcggatc
     1621 aatgacggca tcgagcggcc cgccgctcag tggttcaagc ggtacaacgg caagaccccg
     1681 gagaagggcg gggcacagaa gaccgaagcg ctcaagccca aggcatggct tgagcagacc
     1741 cgcgaggcat gggccgacca tgccaaccgg gcattagagc gggctggcca cgacgcccgc
     1801 attgaccaca gaacacttga ggcgcagggc atcgagcgcc tgcccggtgt tcacctgggg
     1861 ccgaacgtgg tggagatgga aggccggggc atccgcaccg accgggcaga cgtggccctg
     1921 aacatcgaca ccgccaacgc ccagatcatc gacttacagg aataccggga ggcaatagac
     1981 catgaacgca atcgacagag tgaagaaatc cagaggcatc aacgagttag cggagcagat
     2041 cgaaccgctg gcccagagca tggcgacact ggccgacgaa gcccggcagg tcatgagcca
     2101 gacccagcag gccagcgagg cgcaggcggc ggagtggctg aaagcccagc gccagacagg
     2161 ggcggcatgg gtggagctgg ccaaagagtt gcgggaggta gccgccgagg tgagcagcgc
     2221 cgcgcagagc gcccggagcg cgtcgcgggg gtggcactgg aagctatggc taaccgtgat
     2281 gctggcttcc atgatgccta cggtggtgct gctgatcgca tcgttgctct tgctcgacct
     2341 gacgccactg acaaccgagg acggctcgat ctggctgcgc ttggtggccc gatgaagaac
     2401 gacaggactt tgcaggccat aggccgacag ctcaaggcca tgggctgtga gcgcttcgat
     2461 atcggcgtca gggacgccac caccggccag atgatgaacc gggaatggtc agccgccgaa
     2521 gtgctccaga acacgccatg gctcaagcgg atgaatgccc agggcaatga cgtgtatatc
     2581 aggcccgccg agcaggagcg gcatggtctg gtgctggtgg acgacctcag cgagtttgac
     2641 ctggatgaca tgaaagccga gggccgggag cctgccctgg tagtggaaac cagcccgaag
     2701 aactatcagg catgggtcaa ggtggccgac gccgcaggcg gtgaacttcg ggggcagatt
     2761 gcccggacgc tggccagcga gtacgacgcc gacccggcca gcgccgacag ccgccactat
     2821 ggccgcttgg cgggcttcac caaccgcaag gacaagcaca ccacccgcgc cggttatcag
     2881 ccgtgggtgc tgctgcgtga atccaagggc aagaccgcca ccgctggccc ggcgctggtg
     2941 cagcaggctg gccagcagat cgagcaggcc cagcggcagc aggagaaggc ccgcaggctg
     3001 gccagcctcg aactgcccga gcggcagctt agccgccacc ggcgcacggc gctggacgag
     3061 taccgcagcg agatggccgg gctggtcaag cgcttcggtg atgacctcag caagtgcgac
     3121 tttatcgccg cgcagaagct ggccagccgg ggccgcagtg ccgaggaaat cggcaaggcc
     3181 atggccgagg ccagcccagc gctggcagag cgcaagcccg gccacgaagc ggattacatc
     3241 gagcgcaccg tcagcaaggt catgggtctg cccagcgtcc agcttgcgcg ggccgagctg
     3301 gcacgggcac cggcaccccg ccagcgaggc atggacaggg gcgggccaga tttcagcatg
     3361 tagtgcttgc gttggtactc acgcctgtta tactatgagt actcacgcac agaagggggt
     3421 tttatggaat acgaaaaaag cgcttcaggg tcggtctacc tgatcaaaag tgacaagggc
     3481 tattggttgc ccggtggctt tggttatacg tcaaacaagg ccgaggctgg ccgcttttca
     3541 gtcgctgata tggccagcct taaccttgac ggctgcacct tgtccttgtt ccgcgaagac
     3601 aagcctttcg gccccggcaa gtttctcggt gactgatatg aaagaccaaa aggacaagca
     3661 gaccggcgac ctgctggcca gccctgacgc tgtacgccaa gcgcgatatg ccgagcgcat
     3721 gaaggccaaa gggatgcgtc agcgcaagtt ctggctgacc gacgacgaat acgaggcgct
     3781 gcgcgagtgc ctggaagaac tcagagcggc gcagggcggg ggtagtgacc ccgccagcgc
     3841 ctaaccacca actgcctgca aaggaggcaa tcaatggcta cccataagcc tatcaatatt
     3901 ctggaggcgt tcgcagcagc gccgccaccg ctggactacg ttttgcccaa catggtggcc
     3961 ggtacggtcg gggcgctggt gtcgcccggt ggtgccggta aatccatgct ggccctgcaa
     4021 ctggccgcac agattgcagg cgggccggat ctgctggagg tgggcgaact gcccaccggc
     4081 ccggtgatct acctgcccgc cgaagacccg cccaccgcta ttcatcaccg cctgcacgcc
     4141 cttggggcgc acctcagcgc cgaggaacgg caagccgtgg ctgacggcct gctgatccag
     4201 ccgctgatcg gcagcctgcc caacatcatg gccccggagt ggttcgacgg cctcaagcgc
     4261 gccgccgagg gccgccgcct gatggtgctg gacacgctgc gccggttcca catcgaggaa
     4321 gaaaacgcca gcggccccat ggcccaggtc atcggtcgca tggaggccat cgccgccgat
     4381 accgggtgct ctatcgtgtt cctgcaccat gccagcaagg gcgcggccat gatgggcgca
     4441 ggcgaccagc agcaggccag ccggggcagc tcggtactgg tcgataacat ccgctggcag
     4501 tcctacctgt cgagcatgac cagcgccgag gccgaggaat ggggtgtgga cgacgaccag
     4561 cgccggttct tcgtccgctt cggtgtgagc aaggccaact atggcgcacc gttcgctgat
     4621 cggtggttca ggcggcatga cggcggggtg ctcaagcccg ccgtgctgga gaggcagcgc
     4681 aagagcaagg gggtgccccg tggtgaagcc taagaacaag cacagcctca gccacgtccg
     4741 gcacgacccg gcgcactgtc tggcccccgg cctgttccgt gccctcaagc ggggcgagcg
     4801 caagcgcagc aagctggacg tgacgtatga ctacggcgac ggcaagcgga tcgagttcag
     4861 cggcccggag ccgctgggcg ctgatgatct gcgcatcctg caagggctgg tggccatggc
     4921 tgggcctaat ggcctagtgc ttggcccgga acccaagacc gaaggcggac ggcagctccg
     4981 gctgttcctg gaacccaagt gggaggccgt caccgctgat gccatggtgg tcaaaggtag
     5041 ctatcgggcg ctggcaaagg aaatcggggc agaggtcgat agtggtgggg cgctcaagca
     5101 catacaggac tgcatcgagc gcctttggaa ggtatccatc atcgcccaga atggccgcaa
     5161 gcggcagggg tttcggctgc tgtcggagta cgccagcgac gaggcggacg ggcgcctgta
     5221 cgtggccctg aaccccttga tcgcgcaggc cgtcatgggt ggcggccagc atgtgcgcat
     5281 cagcatggac gaggtgcggg cgctggacag cgaaaccgcc cgcctgctgc accagcggct
     5341 gtgtggctgg atcgaccccg gcaaaaccgg caaggcttcc atagatacct tgtgcggcta
     5401 tgtctggccg tcagaggcca gtggttcgac catgcgcaag cgccgccagc gggtgcgcga
     5461 ggcgttgccg gagctggtcg cgctgggctg gacggtaacc gagttcgcgg cgggcaagta
     5521 cgacatcacc cggcccaagg cggcaggctg acccccccca ctctattgta aacaagacat
     5581 ttttatcttt tatattcaat ggcttatttt cctgctaatt ggtaatacca tgaaaaatac
     5641 catgctcaga aaaggcttaa caatattttg aaaaattgcc tactgagcgc tgccgcacag
     5701 ctccatagcg tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac
     5761 tcatcgcagt acggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5821 ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg
     5881 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     5941 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     6001 gccagtgatc aatttttaaa gtatgggcaa tcaattgctc ctgttaaaat tgctttagaa
     6061 atactttggc agcggtttgt tgtattgagt ttcatttgcg cattggttaa atggaaagtg
     6121 acagtacgct cactgcaacc taatattttt gaaatatccc aagagctttt tccttcgcat
     6181 gcccacgcta aacattcttt ttctcttttg gttaaatcgt tgtttgattt attatttgct
     6241 atatttattt ttcgataatt atcaactaga gaaggaacaa ttaatggtat gttcatacac
     6301 gcatgtaaaa ataaactatc tatatagttg tctttttctg aatgtgcaaa actaagcatt
     6361 ccgaagccat tgttagccgt atgaataggg aaactaaacc cagtgataag acctgatgtt
     6421 ttcgcttctt taattacatt tggagatttt ttatttacag cattgttttc aaatatattc
     6481 caattaattg gtgaatgatt ggagttagaa taatctacta taggatcata ttttatcaaa
     6541 ttagcgtcat cataatattg cctccatttt ttagggtaat tatctaggat tgaaatatca
     6601 gatttaacca tagaatgagg ataaatgatc gcgagtaaat aatattcaca atgtaccatt
     6661 ttagtcatat cagataagca ttgattaata tcattattgc ttctacaagc tttaatttta
     6721 ttaattattc tgtatgtgtc gtcggcattt atgtttttca ttctgcccct ccgaaaggct
     6781 taagtcctcg cgtagtacct gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     6841 cgatggcgcg ccgcccaatt attgaaggcc tccctaacgg ggggcctttt tttgtttctg
     6901 gtctcccgct taacgatcgt tggctgacct gtaggatcgt acaggtttac gcaagaaaat
     6961 ggtttgttac agtcgaataa aagctgtcac cggatgtgct ttccggtctg atgagtccgt
     7021 gaggacgaaa cagcctctac aaataatttt gtttaagaac tgggaagagc gcagattaaa
     7081 gaggagaaat actagatggc ttcctccgaa gacgttatca aagagttcat gcgtttcaaa
     7141 gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt
     7201 cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg
     7261 ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac
     7321 ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt
     7381 gttatgaact tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac
     7441 ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt
     7501 atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt
     7561 gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct
     7621 gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc
     7681 gacatcaaac tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa
     7741 cgtgctgaag gtcgtcactc caccggtgct taataacgct gatagtgcta gtgtagatcg
     7801 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7861 gagaggcggt ttgcgtattg ggcgcatgca taaaaactgt tgtaattcat taagcattct
     7921 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     7981 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     8041 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     8101 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     8161 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     8221 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     8281 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     8341 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     8401 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     8461 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     8521 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     8581 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     8641 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     8701 tgcgaagtga tcttccgtca caggtattta ttcgaagacg aaagggcctc gtgatacgcc
     8761 tatttttata ggttaatgtc atgataataa tggtttctta gacgtca
//
