LOCUS       sequence-314607-        8781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RSF1010 origin, CymR(AM) 
            repressor, PCymRC promoter, mRFP, Gentamicin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8781)
  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 8781)
  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: pFCyR5
FEATURES             Location/Qualifiers
     source          1..8781
                     /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"
     protein_bind    6809..6836
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    6867..6894
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     misc_RNA        6898..6947
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             6992..7003
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7010..7687
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(7964..7969,7970..7986,7987..7992)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             8181..8714
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
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 cccaccgcca 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 aacgtttgaa ttttgcataa cgttcacgtg caatttccag ggtgctatta
     6061 cgcacacgtt caaaacgttc tttatctttc tgccacaggc tacgaactac cagaccacga
     6121 acgctgttaa aaatcagcca cagaatatct tcggcatcat cacggctcag accacggctc
     6181 accagcacac ccagccacat atcttcaaca acaaaacgat tacgttcaac ggtacgctga
     6241 ataccttcac gcagtgccgg atcacgatct gctgcaacaa tcagatccag gccgatgcta
     6301 aaatcatcat ccagaaaaaa ttctgctgca tcatccagca tctgctgaat aacatcatct
     6361 tccggtttca gttttgccag acgtgcacgg ctacgttcgg taatctgctc atacagccat
     6421 tcaaaggttg ccagcagcag ttccagtttg gtcggaaaat gatggctctg tgcaccacgg
     6481 ctaacaccgg ctgcacccgg aacatctgca atacgaaaac ctgcataacc tttttcacgc
     6541 agaacaccca gtgctgctgc aatcagttta ccctgggttt ccattgcacg ttctgcctgg
     6601 gtacgacgtt tcgggctcat aagaatctcc gtttggtatt tactatgaat attccattgg
     6661 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggcgcgcc gccccaatta
     6721 ttgaaggcct ccctaacggg gggccttttt ttgtttctgg tctcccgctt aacgatcgtt
     6781 ggctgggata catatgtcaa tgtaccggaa caaacagaca atctggtctg tttgtattat
     6841 ggaaaatttt tctgtataat agattcaaca aacagacaat ctggtctgtt tgtaaagctg
     6901 tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct ctacaaataa
     6961 ttttgtttaa gaactgggaa gagcgcagat taaagaggag aaatactaga tggcttcctc
     7021 cgaagacgtt atcaaagagt tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg
     7081 tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta cccagaccgc
     7141 taaactgaaa gttaccaaag gtggtccgct gccgttcgct tgggacatcc tgtccccgca
     7201 gttccagtac ggttccaaag cttacgttaa acacccggct gacatcccgg actacctgaa
     7261 actgtccttc ccggaaggtt tcaaatggga acgtgttatg aacttcgaag acggtggtgt
     7321 tgttaccgtt acccaggact cctccctgca agacggtgag ttcatctaca aagttaaact
     7381 gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca tgggttggga
     7441 agcttccacc gaacgtatgt acccggaaga cggtgctctg aaaggtgaaa tcaaaatgcg
     7501 tctgaaactg aaagacggtg gtcactacga cgctgaagtt aaaaccacct acatggctaa
     7561 aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc aaactggaca tcacctccca
     7621 caacgaagac tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg
     7681 tgcttaataa cgctgatagt gctagtgtag atcgcgcttt ccagtcggga aacctgtcgt
     7741 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgca
     7801 tgcataaaaa ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     7861 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     7921 ccatggacgc acaccgtgga aacggatgaa ggcacgaacc cagttgacat aagcctgttc
     7981 ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     8041 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     8101 tgtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     8161 gatgttatgg agcagcaacg atgttacgca gcagcaacga tgttacgcag cagggcagtc
     8221 gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt aggctcggcc
     8281 ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag ttcggagacg
     8341 tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg ctccgtagta
     8401 agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct ctcgcggctt
     8461 acgttctgcc caagtttgag cagccgcgta gtgagatcta tatctatgat ctcgcagtct
     8521 ccggagagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc aagcatgagg
     8581 ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac gatcccgcag
     8641 tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat atcgacccaa
     8701 gtaccgccac ctaacaattc gttcaagccg agatcggctt cccggccgcg gagttgttcg
     8761 gtaaattgtc acaacgccgc a
//
