LOCUS       sequence-311959-        8289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RK2 and pUC origin, RhaR-RhaS 
            activator, PrhaBAD promoter, mRFP, Gentamicin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8289)
  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 8289)
  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: pKRR5
FEATURES             Location/Qualifiers
     source          1..8289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(249..1397)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(1669..2040)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2073..2182
                     /note="incP origin of transfer"
     rep_origin      2335..2966
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      3429..4017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3918..3937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4335..4357
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4349..4366
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4350..4366
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4369..5217)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQSIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(5291..6127)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHGVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFLREFNWSPRDIRQGRDGFLQ"
     promoter        6273..6391
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     misc_RNA        6406..6455
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             6500..6511
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6518..7195
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(7472..7477,7478..7494,7495..7500)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             7689..8222
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ggtggcactt ttcggggatc gtggaaacga taggcctatg ccatgcgggt caaggcgact
       61 tccggcaagc tatacgcgcc ctagaattgt caattttaat cctctgttta tcggcagttc
      121 gtagagcgcg ccgtgcgtcc cgagcgatac tgagcgaagc aagtgcgtcg agcagtgccc
      181 gcttgttcct gaaatgccag taaagcgctg gctgctgaac ccccagccgg aactgacccc
      241 acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg ttccaccagg
      301 ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg
      361 gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta cggctcccgg
      421 tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg gtacttctcc
      481 catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc gtcgatcagg
      541 acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg cagcagcgac
      601 accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc
      661 gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc caggtcctgg
      721 caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc
      781 aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg
      841 atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt
      901 ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat cgctcgcatc
      961 gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat ctgctgcttc
     1021 gtgtgtttca gcaacgcggc ctgcttggcc tcgctgacct gttttgccag gtcctcgccg
     1081 gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc
     1141 aaacctgccg cctcctgttc gagacgacgc gaacgctcca cggcggccga tggcgcgggc
     1201 agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc ttgctggacc
     1261 atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg
     1321 gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg
     1381 tcaaacgtcc gattcattca ccctccttgc gggattgccc cgactcacgc cggggcaatg
     1441 tgcccttatt cctgatttga cccgcctggt gccttggtgt ccagataatc caccttatcg
     1501 gcaatgaagt cggtcccgta gaccgtctgg ccgtccttct cgtacttggt attccgaatc
     1561 ttgccctgca cgaataccag ctccgcgaag tcgctcttct tgatggagcg catggggacg
     1621 tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc atggctctgc
     1681 cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct cttcgatctt
     1741 cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag
     1801 ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg
     1861 gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg ccagcaggta
     1921 ggcctacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc gttcgtctgg
     1981 aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt catcagccat
     2041 ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag caggattccc
     2101 gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg
     2161 gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg
     2221 aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc
     2281 tataatgacc ccgaagcagg gttatgcagc ggaaaagatc cgtcgaccct ttccgacgct
     2341 caccgggctg gttgccctcg ccgctgggct ggcggccgtc tatggccctg caaacgcgcc
     2401 agaaacgccg tcgaagccgt gtgcgagaca ccgcggccgc cggcgttgtg gatacctcgc
     2461 ggaaaacttg gccctcactg acagatgagg ggcggacgtt gacacttgag gggccgactc
     2521 acccggcgcg gcgttgacag atgaggggca ggctcgattt cggccggcga cgtggagctg
     2581 gccagcctcg caaatcggcg aaaacgcctg attttacgcg agtttcccac agatgatgtg
     2641 gacaagcctg gggataagtg ccctgcggta ttgacacttg aggggcgcga ctactgacag
     2701 atgaggggcg cgatccttga cacttgaggg gcagagtgct gacagatgag gggcgcacat
     2761 attgacattt gaggggctgt ccacaggcag aaaatccagc atttgcaagg gtttccgcca
     2821 gtttttcggc caccgctaac ctgtctttta acctgctttt aaaccaatat ttataaacct
     2881 tgtttttaac cagggctgcg ccctgtgcgc gtgaccgcgc acgccgaagg ggggtgcccc
     2941 cccttctcga accctcccgg cccgctaacg cgggcctccc atccccccag gggctgcgcc
     3001 cctcggccgc gaacggcctc accccaaaaa tggcagccaa gctgaccact tctgcgctcg
     3061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3241 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3301 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3361 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3541 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3781 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttcg tttaagggca
     4081 ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt acggcctatt
     4141 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4201 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     4261 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     4321 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgatt aatctttctg
     4381 cgaattgaga tgacgccact ggctgggcgt catcccggtt tcccgggtaa acaccaccga
     4441 aaaatagtta ctatcttcaa agccacattc ggtcgaaata tcactgatta acaggcggct
     4501 atgctggaga agatattgcg catgacacac tctgacctgt cgcagatatt gattgatggt
     4561 cattccagtc tgctggcgaa attgctgacg caaaacgcgc tcactgcacg atgcctcatc
     4621 acaaaattta tccagcgcaa agggactttt caggctagcc gccagccggg taatcagctt
     4681 atccagcaac gtttcgctgg atgttggcgg caacgaatca ctggtgtaac gatggcgatt
     4741 cagcaacatc accaactgcc cgaacagcaa ctcagccatt tcgttagcaa acggcacatg
     4801 ctgactactt tcatgctcaa gctgaccgat aacctgccgc gcctgcgcca tccccatgct
     4861 acctaagcgc cagtgtggtt gccctgcgct tgcgttaaat cccggaatcg ccccctgcca
     4921 gtcaagattc agcttcagac gctccgggca ataaataata ctctgcaaaa ccagatcgtt
     4981 aacggaagcg taggagtgtt tatcgtcagc atgaatgtaa aagagatcgc cacgggtaat
     5041 gcgataaggg cgatcgttga gtacatgcag gccattaccg cgccagacaa tcaccagctc
     5101 acaaaaatca tgtgtatgtt cagcaaagac atcttgcgga taacggtcag ccacagcgac
     5161 tgcctgctgg tcgctggcaa aaaaatcatc tttgagaagt tttaactgat gcgccaccgt
     5221 ggctacctcg gccagagaac gaagttgatt attcgcaata tggcgtacaa atacgttgag
     5281 aagattcgcg ttattgcaga aagccatccc gtccctggcg aatatcacgc ggtgaccagt
     5341 taaactctcg gagaaaaagc gtcgaaaagt ggttactgtc gctgaatcca cagcgatagg
     5401 cgatgtcagt aacgctggcc tcgctgtggc gtagcagatg tcgggctttc atcagtcgca
     5461 ggcggttcag gtatcgctga ggcgtcagtc ccgtttgctg cttaagctgc cgatgtagcg
     5521 tacgcagtga aagagaaaat tgatccgcca cggcatccca attcacctca tcggcaaaat
     5581 ggtcctccag ccaggccaga agcaagttga gacgtgatgc gctgttttcc aggttctcct
     5641 gcaaactgct tttacgcagc aagagcagta attgcataaa caggatctcg cgactggcgg
     5701 tcgagggtaa atcattttcc ccttcctgct gttccatctg tgcaaccagc tgtcgcacct
     5761 gctgcaatac gccgtggtta acgcgccagt gagacggata ctgcccatcc agctcttgtg
     5821 gcagcaactg attcagcccg gcgagaaact gaaatcgatc cggcgagcga tacagcacat
     5881 tggtcagaca cagattatcg gtatgttcat acagatgccg atcatgatcg cgtacgaaac
     5941 agaccgtgcc accggtgatg gtatagggct gcccattaaa cacatgaata cccgtgccat
     6001 gttcgacaat cacaatttca tgaaaatcat gatgatgttc aggaaaatcc gcctgcggga
     6061 gccggggttc tatcgccacg gacgcgttac cagacggaaa aaaatccaca ctatgtaata
     6121 cggtcatact ggcttcctga tgtcgtcaac acggcgaaat agtaatcacg aggtcaggtt
     6181 cttaccttaa attttcgacg gaaaaccacg taaaaaacgt cgatttttca agatacagcg
     6241 tgaattttca ggaaatgcgg tgagcatcac atcaccacaa ttcagcaaat tgtgaacatc
     6301 atcacgttca tctttccctg gttgccaatg gcccattttc ctgtcagtaa cgagaaggtc
     6361 gcgaattcag gcgcttttta gactggtcgt aatgaaattc agcagctgtc accggatgtg
     6421 ctttccggtc tgatgagtcc gtgaggacga aacagcctct acaaataatt ttgtttaaga
     6481 actgggaaga gcgcagatta aagaggagaa atactagatg gcttcctccg aagacgttat
     6541 caaagagttc atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga
     6601 aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt
     6661 taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg
     6721 ttccaaagct tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc
     6781 ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac
     6841 ccaggactcc tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa
     6901 cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga
     6961 acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa
     7021 agacggtggt cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca
     7081 gctgccgggt gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta
     7141 caccatcgtt gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataacg
     7201 ctgatagtgc tagtgtagat cgcgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7261 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgcatg cataaaaact
     7321 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
     7381 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggacgcac
     7441 accgtggaaa cggatgaagg cacgaaccca gttgacataa gcctgttcgg ttcgtaaact
     7501 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     7561 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg tacagtctat
     7621 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     7681 cagcaacgat gttacgcagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     7741 agttaggtgg ctcaagtatg ggcatcattc gcacatgtag gctcggccct gaccaagtca
     7801 aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt cggagacgta gccacctact
     7861 cccaacatca gccggactcc gattacctcg ggaacttgct ccgtagtaag acattcatcg
     7921 cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct cgcggcttac gttctgccca
     7981 agtttgagca gccgcgtagt gagatctata tctatgatct cgcagtctcc ggagagcacc
     8041 ggaggcaggg cattgccacc gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg
     8101 gtgcttatgt gatctacgtg caagcagatt acggtgacga tcccgcagtg gctctctata
     8161 caaagttggg catacgggaa gaagtgatgc actttgatat cgacccaagt accgccacct
     8221 aacaattcgt tcaagccgag atcggcttcc cggccgcgga gttgttcggt aaattgtcac
     8281 aacgccgca
//
