LOCUS       sequence-332867-        9050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RSF1010 origin, LuxR activator, 
            PLuxB promoter, mRFP, Carbenicillin Resistance Marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9050)
  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 9050)
  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: pFLxR4
FEATURES             Location/Qualifiers
     source          1..9050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        261..310
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             355..366
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             373..1050
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        1285..1356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1357..1425,1426..2217)
                     /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     complement(1575..1594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2657..3051
                     /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            3393..3480
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             4719..5690
                     /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             6201..7040
                     /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             7027..7878
                     /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     8302..8324
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8316..8333
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8317..8333
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attattctgt atgtgtcgtc ggcatttatg tttttcattc tgcccctccg aaaggcttaa
       61 gtcctcgcgt agtacctggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
      121 tggcgcgccg cccaattatt gaaggcctcc ctaacggggg gccttttttt gtttctggtc
      181 tcccgcttaa cgatcgttgg ctgacctgta ggatcgtaca ggtttacgca agaaaatggt
      241 ttgttacagt cgaataaaag ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag
      301 gacgaaacag cctctacaaa taattttgtt taagaactgg gaagagcgca gattaaagag
      361 gagaaatact agatggcttc ctccgaagac gttatcaaag agttcatgcg tttcaaagtt
      421 cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt
      481 ccgtacgaag gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc
      541 gcttgggaca tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg
      601 gctgacatcc cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt
      661 atgaacttcg aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt
      721 gagttcatct acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg
      781 cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct
      841 ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa
      901 gttaaaacca cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac
      961 atcaaactgg acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt
     1021 gctgaaggtc gtcactccac cggtgcttaa taacgctgat agtgctagtg tagatcgcgc
     1081 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1141 aggcggtttg cgtattgggc gcatgcataa aaactgttgt aattcattaa gcattctgcc
     1201 gacatggaag ccatcacaaa ctgcatgatg aacctgaatc gccagcggca tcagcacctt
     1261 gtcgccttgc gtataatatt tgcccattca aatatgtatc cgctcatgag acaataaccc
     1321 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1381 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1441 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1501 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1561 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1621 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1681 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1741 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1801 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1861 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     1921 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     1981 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2041 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2101 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2161 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2221 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2281 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccaggt ggcacttttc
     2341 ggggatcttc aaattcccgt tgcacatagc ccggcaattc ctttccctgc tctgccataa
     2401 gcgcagcgaa tgccgggtaa tactcgtcaa cgatctgata gagaagggtt tgctcgggtc
     2461 ggtggctctg gtaacgacca gtatcccgat cccggctggc cgtcctggcc gccacatgag
     2521 gcatgttccg cgtccttgca atactgtgtt tacatacagt ctatcgctta gcggaaagtt
     2581 cttttaccct cagccgaaat gcctgccgtt gctagacatt gccagccagt gcccgtcact
     2641 cccgtactaa ctgtcacgaa cccctgcaat aactgtcacg cccccctgca ataactgtca
     2701 cgaacccctg caataactgt cacgccccca aacctgcaaa cccagcaggg gcgggggctg
     2761 gcggggtgtt ggaaaaatcc atccatgatt atctaagaat aatccactag gcgcggttat
     2821 cagcgccctt gtggggcgct gctgcccttg cccaatatgc ccggccagag gccggatagc
     2881 tggtctattc gctgcgctag gctacacacc gccccaccgc tgcgcggcag ggggaaaggc
     2941 gggcaaagcc cgctaaaccc cacaccaaac cccgcagaaa tacgctggag cgcttttagc
     3001 cgctttagcg gcctttcccc ctacccgaag ggtgggggcg cgtgtgcagc cccgcagggc
     3061 ctgtctcggt cgatcattca gcccggctca tccttctggc gtggcggcag accgaacaag
     3121 gcgcggtcgt ggtcgcgttc aaggtacgca tccattgccg ccatgagccg atcctccggc
     3181 cactcgctgc tgttcacctt ggccaaaatc atggccccca ccagcacctt gcgccttgtt
     3241 tcgttcttgc gctcttgctg ctgttccctt gcccgcaccc gctgaatttc ggcattgatt
     3301 cgcgctcgtt gttcttcgag cttggccagc cgatccgccg ccttgttgct ccccttaacc
     3361 atcttgacac cccattgtta atgtgctgtc tcgtaggcta tcatggaggc acagcggcgg
     3421 caatcccgac cctactttgt aggggagggc gcacttaccg gtttctcttc gagaaactgg
     3481 cctaacggcc acccttcggg cggtgcgctc tccgagggcc attgcatgga gccgaaaagc
     3541 aaaagcaaca gcgaggcagc atggcgattt atcaccttac ggcgaaaacc ggcagcaggt
     3601 cgggcggcca atcggccagg gccaaggccg actacatcca gcgcgaaggc aagtatgccc
     3661 gcgacatgga tgaagtcttg cacgccgaat ccgggcacat gccggagttc gtcgagcggc
     3721 ccgccgacta ctgggatgct gccgacctgt atgaacgcgc caatgggcgg ctgttcaagg
     3781 aggtcgaatt tgccctgccg gtcgagctga ccctcgacca gcagaaggcg ctggcgtccg
     3841 agttcgccca gcacctgacc ggtgccgagc gcctgccgta tacgctggcc atccatgccg
     3901 gtggcggcga gaacccgcac tgccacctga tgatctccga gcggatcaat gacggcatcg
     3961 agcggcccgc cgctcagtgg ttcaagcggt acaacggcaa gaccccggag aagggcgggg
     4021 cacagaagac cgaagcgctc aagcccaagg catggcttga gcagacccgc gaggcatggg
     4081 ccgaccatgc caaccgggca ttagagcggg ctggccacga cgcccgcatt gaccacagaa
     4141 cacttgaggc gcagggcatc gagcgcctgc ccggtgttca cctggggccg aacgtggtgg
     4201 agatggaagg ccggggcatc cgcaccgacc gggcagacgt ggccctgaac atcgacaccg
     4261 ccaacgccca gatcatcgac ttacaggaat accgggaggc aatagaccat gaacgcaatc
     4321 gacagagtga agaaatccag aggcatcaac gagttagcgg agcagatcga accgctggcc
     4381 cagagcatgg cgacactggc cgacgaagcc cggcaggtca tgagccagac ccagcaggcc
     4441 agcgaggcgc aggcggcgga gtggctgaaa gcccagcgcc agacaggggc ggcatgggtg
     4501 gagctggcca aagagttgcg ggaggtagcc gccgaggtga gcagcgccgc gcagagcgcc
     4561 cggagcgcgt cgcgggggtg gcactggaag ctatggctaa ccgtgatgct ggcttccatg
     4621 atgcctacgg tggtgctgct gatcgcatcg ttgctcttgc tcgacctgac gccactgaca
     4681 accgaggacg gctcgatctg gctgcgcttg gtggcccgat gaagaacgac aggactttgc
     4741 aggccatagg ccgacagctc aaggccatgg gctgtgagcg cttcgatatc ggcgtcaggg
     4801 acgccaccac cggccagatg atgaaccggg aatggtcagc cgccgaagtg ctccagaaca
     4861 cgccatggct caagcggatg aatgcccagg gcaatgacgt gtatatcagg cccgccgagc
     4921 aggagcggca tggtctggtg ctggtggacg acctcagcga gtttgacctg gatgacatga
     4981 aagccgaggg ccgggagcct gccctggtag tggaaaccag cccgaagaac tatcaggcat
     5041 gggtcaaggt ggccgacgcc gcaggcggtg aacttcgggg gcagattgcc cggacgctgg
     5101 ccagcgagta cgacgccgac ccggccagcg ccgacagccg ccactatggc cgcttggcgg
     5161 gcttcaccaa ccgcaaggac aagcacacca cccgcgccgg ttatcagccg tgggtgctgc
     5221 tgcgtgaatc caagggcaag accgccaccg ctggcccggc gctggtgcag caggctggcc
     5281 agcagatcga gcaggcccag cggcagcagg agaaggcccg caggctggcc agcctcgaac
     5341 tgcccgagcg gcagcttagc cgccaccggc gcacggcgct ggacgagtac cgcagcgaga
     5401 tggccgggct ggtcaagcgc ttcggtgatg acctcagcaa gtgcgacttt atcgccgcgc
     5461 agaagctggc cagccggggc cgcagtgccg aggaaatcgg caaggccatg gccgaggcca
     5521 gcccagcgct ggcagagcgc aagcccggcc acgaagcgga ttacatcgag cgcaccgtca
     5581 gcaaggtcat gggtctgccc agcgtccagc ttgcgcgggc cgagctggca cgggcaccgg
     5641 caccccgcca gcgaggcatg gacaggggcg ggccagattt cagcatgtag tgcttgcgtt
     5701 ggtactcacg cctgttatac tatgagtact cacgcacaga agggggtttt atggaatacg
     5761 aaaaaagcgc ttcagggtcg gtctacctga tcaaaagtga caagggctat tggttgcccg
     5821 gtggctttgg ttatacgtca aacaaggccg aggctggccg cttttcagtc gctgatatgg
     5881 ccagccttaa ccttgacggc tgcaccttgt ccttgttccg cgaagacaag cctttcggcc
     5941 ccggcaagtt tctcggtgac tgatatgaaa gaccaaaagg acaagcagac cggcgacctg
     6001 ctggccagcc ctgacgctgt acgccaagcg cgatatgccg agcgcatgaa ggccaaaggg
     6061 atgcgtcagc gcaagttctg gctgaccgac gacgaatacg aggcgctgcg cgagtgcctg
     6121 gaagaactca gagcggcgca gggcgggggt agtgaccccg ccagcgccta accaccaact
     6181 gcctgcaaag gaggcaatca atggctaccc ataagcctat caatattctg gaggcgttcg
     6241 cagcagcgcc gccaccgctg gactacgttt tgcccaacat ggtggccggt acggtcgggg
     6301 cgctggtgtc gcccggtggt gccggtaaat ccatgctggc cctgcaactg gccgcacaga
     6361 ttgcaggcgg gccggatctg ctggaggtgg gcgaactgcc caccggcccg gtgatctacc
     6421 tgcccgccga agacccgccc accgctattc atcaccgcct gcacgccctt ggggcgcacc
     6481 tcagcgccga ggaacggcaa gccgtggctg acggcctgct gatccagccg ctgatcggca
     6541 gcctgcccaa catcatggcc ccggagtggt tcgacggcct caagcgcgcc gccgagggcc
     6601 gccgcctgat ggtgctggac acgctgcgcc ggttccacat cgaggaagaa aacgccagcg
     6661 gccccatggc ccaggtcatc ggtcgcatgg aggccatcgc cgccgatacc gggtgctcta
     6721 tcgtgttcct gcaccatgcc agcaagggcg cggccatgat gggcgcaggc gaccagcagc
     6781 aggccagccg gggcagctcg gtactggtcg ataacatccg ctggcagtcc tacctgtcga
     6841 gcatgaccag cgccgaggcc gaggaatggg gtgtggacga cgaccagcgc cggttcttcg
     6901 tccgcttcgg tgtgagcaag gccaactatg gcgcaccgtt cgctgatcgg tggttcaggc
     6961 ggcatgacgg cggggtgctc aagcccgccg tgctggagag gcagcgcaag agcaaggggg
     7021 tgccccgtgg tgaagcctaa gaacaagcac agcctcagcc acgtccggca cgacccggcg
     7081 cactgtctgg cccccggcct gttccgtgcc ctcaagcggg gcgagcgcaa gcgcagcaag
     7141 ctggacgtga cgtatgacta cggcgacggc aagcggatcg agttcagcgg cccggagccg
     7201 ctgggcgctg atgatctgcg catcctgcaa gggctggtgg ccatggctgg gcctaatggc
     7261 ctagtgcttg gcccggaacc caagaccgaa ggcggacggc agctccggct gttcctggaa
     7321 cccaagtggg aggccgtcac cgctgatgcc atggtggtca aaggtagcta tcgggcgctg
     7381 gcaaaggaaa tcggggcaga ggtcgatagt ggtggggcgc tcaagcacat acaggactgc
     7441 atcgagcgcc tttggaaggt atccatcatc gcccagaatg gccgcaagcg gcaggggttt
     7501 cggctgctgt cggagtacgc cagcgacgag gcggacgggc gcctgtacgt ggccctgaac
     7561 cccttgatcg cgcaggccgt catgggtggc ggccagcatg tgcgcatcag catggacgag
     7621 gtgcgggcgc tggacagcga aaccgcccgc ctgctgcacc agcggctgtg tggctggatc
     7681 gaccccggca aaaccggcaa ggcttccata gataccttgt gcggctatgt ctggccgtca
     7741 gaggccagtg gttcgaccat gcgcaagcgc cgccagcggg tgcgcgaggc gttgccggag
     7801 ctggtcgcgc tgggctggac ggtaaccgag ttcgcggcgg gcaagtacga catcacccgg
     7861 cccaaggcgg caggctgacc ccccccactc tattgtaaac aagacatttt tatcttttat
     7921 attcaatggc ttattttcct gctaattggt aataccatga aaaataccat gctcagaaaa
     7981 ggcttaacaa tattttgaaa aattgcctac tgagcgctgc cgcacagctc catagcgttt
     8041 aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagtacg
     8101 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     8161 ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     8221 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     8281 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgatcaat
     8341 ttttaaagta tgggcaatca attgctcctg ttaaaattgc tttagaaata ctttggcagc
     8401 ggtttgttgt attgagtttc atttgcgcat tggttaaatg gaaagtgaca gtacgctcac
     8461 tgcaacctaa tatttttgaa atatcccaag agctttttcc ttcgcatgcc cacgctaaac
     8521 attctttttc tcttttggtt aaatcgttgt ttgatttatt atttgctata tttatttttc
     8581 gataattatc aactagagaa ggaacaatta atggtatgtt catacacgca tgtaaaaata
     8641 aactatctat atagttgtct ttttctgaat gtgcaaaact aagcattccg aagccattgt
     8701 tagccgtatg aatagggaaa ctaaacccag tgataagacc tgatgttttc gcttctttaa
     8761 ttacatttgg agatttttta tttacagcat tgttttcaaa tatattccaa ttaattggtg
     8821 aatgattgga gttagaataa tctactatag gatcatattt tatcaaatta gcgtcatcat
     8881 aatattgcct ccatttttta gggtaattat ctaggattga aatatcagat ttaaccatag
     8941 aatgaggata aatgatcgcg agtaaataat attcacaatg taccatttta gtcatatcag
     9001 ataagcattg attaatatca ttattgcttc tacaagcttt aattttatta
//
