LOCUS       sequence-314923-        8859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RSF1010 origin, VanR(AM) 
            repressor, PVanCC promoter, mRFP, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8859)
  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 8859)
  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: pFVR5
FEATURES             Location/Qualifiers
     source          1..8859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      456..850
                     /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            1192..1279
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2518..3489
                     /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             4000..4839
                     /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             4826..5677
                     /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     6101..6123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6115..6132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6116..6132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        7102..7151
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             7196..7207
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7214..7891
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(8168..8173,8174..8190,8191..8196)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(8385..8859,1..59)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac
       61 aattcgttca agccgagatc ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac
      121 gccgcaggtg gcacttttcg gggatcttca aattcccgtt gcacatagcc cggcaattcc
      181 tttccctgct ctgccataag cgcagcgaat gccgggtaat actcgtcaac gatctgatag
      241 agaagggttt gctcgggtcg gtggctctgg taacgaccag tatcccgatc ccggctggcc
      301 gtcctggccg ccacatgagg catgttccgc gtccttgcaa tactgtgttt acatacagtc
      361 tatcgcttag cggaaagttc ttttaccctc agccgaaatg cctgccgttg ctagacattg
      421 ccagccagtg cccgtcactc ccgtactaac tgtcacgaac ccctgcaata actgtcacgc
      481 ccccctgcaa taactgtcac gaacccctgc aataactgtc acgcccccaa acctgcaaac
      541 ccagcagggg cgggggctgg cggggtgttg gaaaaatcca tccatgatta tctaagaata
      601 atccactagg cgcggttatc agcgcccttg tggggcgctg ctgcccttgc ccaatatgcc
      661 cggccagagg ccggatagct ggtctattcg ctgcgctagg ctacacaccg ccccaccgct
      721 gcgcggcagg gggaaaggcg ggcaaagccc gctaaacccc acaccaaacc ccgcataaat
      781 acgctggagc gcttttagcc gctttagcgg cctttccccc tacccgaagg gtgggggcgc
      841 gtgtgcagcc ccgcagggcc tgtctcggtc gatcattcag cccggctcat ccttctggcg
      901 tggcggcaga ccgaacaagg cgcggtcgtg gtcgcgttca aggtacgcat ccattgccgc
      961 catgagccga tcctccggcc actcgctgct gttcaccttg gccaaaatca tggcccccac
     1021 cagcaccttg cgccttgttt cgttcttgcg ctcttgctgc tgttcccttg cccgcacccg
     1081 ctgaatttcg gcattgattc gcgctcgttg ttcttcgagc ttggccagcc gatccgccgc
     1141 cttgttgctc cccttaacca tcttgacacc ccattgttaa tgtgctgtct cgtaggctat
     1201 catggaggca cagcggcggc aatcccgacc ctactttgta ggggagggcg cacttaccgg
     1261 tttctcttcg agaaactggc ctaacggcca cccttcgggc ggtgcgctct ccgagggcca
     1321 ttgcatggag ccgaaaagca aaagcaacag cgaggcagca tggcgattta tcaccttacg
     1381 gcgaaaaccg gcagcaggtc gggcggccaa tcggccaggg ccaaggccga ctacatccag
     1441 cgcgaaggca agtatgcccg cgacatggat gaagtcttgc acgccgaatc cgggcacatg
     1501 ccggagttcg tcgagcggcc cgccgactac tgggatgctg ccgacctgta tgaacgcgcc
     1561 aatgggcggc tgttcaagga ggtcgaattt gccctgccgg tcgagctgac cctcgaccag
     1621 cagaaggcgc tggcgtccga gttcgcccag cacctgaccg gtgccgagcg cctgccgtat
     1681 acgctggcca tccatgccgg tggcggcgag aacccgcact gccacctgat gatctccgag
     1741 cggatcaatg acggcatcga gcggcccgcc gctcagtggt tcaagcggta caacggcaag
     1801 accccggaga agggcggggc acagaagacc gaagcgctca agcccaaggc atggcttgag
     1861 cagacccgcg aggcatgggc cgaccatgcc aaccgggcat tagagcgggc tggccacgac
     1921 gcccgcattg accacagaac acttgaggcg cagggcatcg agcgcctgcc cggtgttcac
     1981 ctggggccga acgtggtgga gatggaaggc cggggcatcc gcaccgaccg ggcagacgtg
     2041 gccctgaaca tcgacaccgc caacgcccag atcatcgact tacaggaata ccgggaggca
     2101 atagaccatg aacgcaatcg acagagtgaa gaaatccaga ggcatcaacg agttagcgga
     2161 gcagatcgaa ccgctggccc agagcatggc gacactggcc gacgaagccc ggcaggtcat
     2221 gagccagacc cagcaggcca gcgaggcgca ggcggcggag tggctgaaag cccagcgcca
     2281 gacaggggcg gcatgggtgg agctggccaa agagttgcgg gaggtagccg ccgaggtgag
     2341 cagcgccgcg cagagcgccc ggagcgcgtc gcgggggtgg cactggaagc tatggctaac
     2401 cgtgatgctg gcttccatga tgcctacggt ggtgctgctg atcgcatcgt tgctcttgct
     2461 cgacctgacg ccactgacaa ccgaggacgg ctcgatctgg ctgcgcttgg tggcccgatg
     2521 aagaacgaca ggactttgca ggccataggc cgacagctca aggccatggg ctgtgagcgc
     2581 ttcgatatcg gcgtcaggga cgccaccacc ggccagatga tgaaccggga atggtcagcc
     2641 gccgaagtgc tccagaacac gccatggctc aagcggatga atgcccaggg caatgacgtg
     2701 tatatcaggc ccgccgagca ggagcggcat ggtctggtgc tggtggacga cctcagcgag
     2761 tttgacctgg atgacatgaa agccgagggc cgggagcctg ccctggtagt ggaaaccagc
     2821 ccgaagaact atcaggcatg ggtcaaggtg gccgacgccg caggcggtga acttcggggg
     2881 cagattgccc ggacgctggc cagcgagtac gacgccgacc cggccagcgc cgacagccgc
     2941 cactatggcc gcttggcggg cttcaccaac cgcaaggaca agcacaccac ccgcgccggt
     3001 tatcagccgt gggtgctgct gcgtgaatcc aagggcaaga ccgccaccgc tggcccggcg
     3061 ctggtgcagc aggctggcca gcagatcgag caggcccagc ggcagcagga gaaggcccgc
     3121 aggctggcca gcctcgaact gcccgagcgg cagcttagcc gccaccggcg cacggcgctg
     3181 gacgagtacc gcagcgagat ggccgggctg gtcaagcgct tcggtgatga cctcagcaag
     3241 tgcgacttta tcgccgcgca gaagctggcc agccggggcc gcagtgccga ggaaatcggc
     3301 aaggccatgg ccgaggccag cccagcgctg gcagagcgca agcccggcca cgaagcggat
     3361 tacatcgagc gcaccgtcag caaggtcatg ggtctgccca gcgtccagct tgcgcgggcc
     3421 gagctggcac gggcaccggc accccgccag cgaggcatgg acaggggcgg gccagatttc
     3481 agcatgtagt gcttgcgttg gtactcacgc ctgttatact atgagtactc acgcacagaa
     3541 gggggtttta tggaatacga aaaaagcgct tcagggtcgg tctacctgat caaaagtgac
     3601 aagggctatt ggttgcccgg tggctttggt tatacgtcaa acaaggccga ggctggccgc
     3661 ttttcagtcg ctgatatggc cagccttaac cttgacggct gcaccttgtc cttgttccgc
     3721 gaagacaagc ctttcggccc cggcaagttt ctcggtgact gatatgaaag accaaaagga
     3781 caagcagacc ggcgacctgc tggccagccc tgacgctgta cgccaagcgc gatatgccga
     3841 gcgcatgaag gccaaaggga tgcgtcagcg caagttctgg ctgaccgacg acgaatacga
     3901 ggcgctgcgc gagtgcctgg aagaactcag agcggcgcag ggcgggggta gtgaccccgc
     3961 cagcgcctaa ccaccaactg cctgcaaagg aggcaatcaa tggctaccca taagcctatc
     4021 aatattctgg aggcgttcgc agcagcgccg ccaccgctgg actacgtttt gcccaacatg
     4081 gtggccggta cggtcggggc gctggtgtcg cccggtggtg ccggtaaatc catgctggcc
     4141 ctgcaactgg ccgcacagat tgcaggcggg ccggatctgc tggaggtggg cgaactgccc
     4201 accggcccgg tgatctacct gcccgccgaa gacccgccca ccgctattca tcaccgcctg
     4261 cacgcccttg gggcgcacct cagcgccgag gaacggcaag ccgtggctga cggcctgctg
     4321 atccagccgc tgatcggcag cctgcccaac atcatggccc cggagtggtt cgacggcctc
     4381 aagcgcgccg ccgagggccg ccgcctgatg gtgctggaca cgctgcgccg gttccacatc
     4441 gaggaagaaa acgccagcgg ccccatggcc caggtcatcg gtcgcatgga ggccatcgcc
     4501 gccgataccg ggtgctctat cgtgttcctg caccatgcca gcaagggcgc ggccatgatg
     4561 ggcgcaggcg accagcagca ggccagccgg ggcagctcgg tactggtcga taacatccgc
     4621 tggcagtcct acctgtcgag catgaccagc gccgaggccg aggaatgggg tgtggacgac
     4681 gaccagcgcc ggttcttcgt ccgcttcggt gtgagcaagg ccaactatgg cgcaccgttc
     4741 gctgatcggt ggttcaggcg gcatgacggc ggggtgctca agcccgccgt gctggagagg
     4801 cagcgcaaga gcaagggggt gccccgtggt gaagcctaag aacaagcaca gcctcagcca
     4861 cgtccggcac gacccggcgc actgtctggc ccccggcctg ttccgtgccc tcaagcgggg
     4921 cgagcgcaag cgcagcaagc tggacgtgac gtatgactac ggcgacggca agcggatcga
     4981 gttcagcggc ccggagccgc tgggcgctga tgatctgcgc atcctgcaag ggctggtggc
     5041 catggctggg cctaatggcc tagtgcttgg cccggaaccc aagaccgaag gcggacggca
     5101 gctccggctg ttcctggaac ccaagtggga ggccgtcacc gctgatgcca tggtggtcaa
     5161 aggtagctat cgggcgctgg caaaggaaat cggggcagag gtcgatagtg gtggggcgct
     5221 caagcacata caggactgca tcgagcgcct ttggaaggta tccatcatcg cccagaatgg
     5281 ccgcaagcgg caggggtttc ggctgctgtc ggagtacgcc agcgacgagg cggacgggcg
     5341 cctgtacgtg gccctgaacc ccttgatcgc gcaggccgtc atgggtggcg gccagcatgt
     5401 gcgcatcagc atggacgagg tgcgggcgct ggacagcgaa accgcccgcc tgctgcacca
     5461 gcggctgtgt ggctggatcg accccggcaa aaccggcaag gcttccatag ataccttgtg
     5521 cggctatgtc tggccgtcag aggccagtgg ttcgaccatg cgcaagcgcc gccagcgggt
     5581 gcgcgaggcg ttgccggagc tggtcgcgct gggctggacg gtaaccgagt tcgcggcggg
     5641 caagtacgac atcacccggc ccaaggcggc aggctgaccc cccccactct attgtaaaca
     5701 agacattttt atcttttata ttcaatggct tattttcctg ctaattggta ataccatgaa
     5761 aaataccatg ctcagaaaag gcttaacaat attttgaaaa attgcctact gagcgctgcc
     5821 gcacagctcc atagcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     5881 tgccactcat cgcagtacgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5941 cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac
     6001 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     6061 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     6121 acgacggcca gtgaaacaga cgaagaatcc atggtcaatc tgcacgaatt gaccatgctg
     6181 cacccagcgg tgcgcctgcg cttgctgctg cttcaaaaac ttttgcatta cgaattgctg
     6241 ccagtgcatg atcacgcata atacgttctg caccttcggc atcaccacag ctaactgcat
     6301 ccagaactgc ctgatgctga cgatgtgctg ccagcagatg ttcatattcg gcagacaggt
     6361 ccatcagatc cagggccagt gcaccggctg ctgcaaacgg ttcaaaacca ttacgtgcca
     6421 gtgcgctttc aactgcacca ttacctgctg cgctaaccag ggtatcatga aatgcctgat
     6481 tatatgcggc ataacgatcc agatcttcac cattcaggcg accggctgca aacagtgctt
     6541 caccttctgc aatcagtaca acaaaacgtg catgggtttc tgaggtcata ccacgttctg
     6601 ccagacgacg tgctgcaaaa ccttccagaa caccacgaac ttcaattgca tcacgaatct
     6661 gatcgctgct aacaccacgg gctgcataac cacgtgcacc cagacgaaca accagacctt
     6721 cttgttccag tgaacgcagt gcgatacgaa ccggcatacg gctaacaccc agtgctgctg
     6781 cggtcggaat ttctgcaata cgttcaccac ttttgatttc accgcttgca atcattttac
     6841 gcagtgccat cataacacgc tgacccggtt taatacgagg catgtccatg tctatcctcc
     6901 ttatttacgt tcgttagttt aagcgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     6961 cattcgatgg cgcgccgcca attattgaag gcctccctaa cggggggcct ttttttgttt
     7021 ctggtctccc gcttaacgat cgttggctga ttggatccaa ttgacagcta gctcagtcct
     7081 aggtaccatt ggatccaata gctgtcaccg gatgtgcttt ccggtctgat gagtccgtga
     7141 ggacgaaaca gcctctacaa ataattttgt ttaagaactg ggaagagcgc agattaaaga
     7201 ggagaaatac tagatggctt cctccgaaga cgttatcaaa gagttcatgc gtttcaaagt
     7261 tcgtatggaa ggttccgtta acggtcacga gttcgaaatc gaaggtgaag gtgaaggtcg
     7321 tccgtacgaa ggtacccaga ccgctaaact gaaagttacc aaaggtggtc cgctgccgtt
     7381 cgcttgggac atcctgtccc cgcagttcca gtacggttcc aaagcttacg ttaaacaccc
     7441 ggctgacatc ccggactacc tgaaactgtc cttcccggaa ggtttcaaat gggaacgtgt
     7501 tatgaacttc gaagacggtg gtgttgttac cgttacccag gactcctccc tgcaagacgg
     7561 tgagttcatc tacaaagtta aactgcgtgg taccaacttc ccgtccgacg gtccggttat
     7621 gcagaaaaaa accatgggtt gggaagcttc caccgaacgt atgtacccgg aagacggtgc
     7681 tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac ggtggtcact acgacgctga
     7741 agttaaaacc acctacatgg ctaaaaaacc ggttcagctg ccgggtgctt acaaaaccga
     7801 catcaaactg gacatcacct cccacaacga agactacacc atcgttgaac agtacgaacg
     7861 tgctgaaggt cgtcactcca ccggtgctta ataacgctga tagtgctagt gtagatcgcg
     7921 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     7981 gaggcggttt gcgtattggg cgcatgcata aaaactgttg taattcatta agcattctgc
     8041 cgacatggaa tccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     8101 tgtcgccttg cgtataatat ttgcccatgg acgcacaccg tggaaacgga tgaaggcacg
     8161 aacccagttg acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc
     8221 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     8281 catggcttgt tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc
     8341 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca
     8401 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca
     8461 tcattcgcac atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc
     8521 ttttcggtcg tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt
     8581 acctcgggaa cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag
     8641 cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga
     8701 tctatatcta tgatctcgca gtctccggag agcaccggag gcagggcatt gccaccgcgc
     8761 tcatcaatct cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag
     8821 cagattacgg tgacgatccc gcagtggctc tctatacaa
//
