LOCUS       sequence-311916-        7398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RK2 and pUC origin, CinR(AM) 
            activator, PCin promoter, mRFP, Gentamicin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7398)
  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 7398)
  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: pKCiR5
FEATURES             Location/Qualifiers
     source          1..7398
                     /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"
     misc_RNA        5515..5564
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             5609..5620
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5627..6304
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(6581..6586,6587..6603,6604..6609)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6798..7331
                     /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 gggcgcacct
     2761 attgacattt gaggggctgt ccacaggcag aaaatccagc atttgcaagg gtttccgccc
     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 gccagtgact attaccaatt
     4381 acgtcgcgtc atgcggatcc tatagggatt gatgatgcgc aattgaacag cccgcgacgc
     4441 ggcggccgag atcgtggtgc agccgagcct gaagcgggcg gttttcaggt aatcgcgtgt
     4501 ggtatgctct gatatgccca ggatgaccga aatatccttg taatccttgc cgagggcggt
     4561 ccagtgcaga cactcgatct cgcgcggcga caatgccggc accggatcgt tttcgccatg
     4621 cagctcatat acggccttgc ggtggatcag atgggcgatc tcgatccact cattgcggca
     4681 gcggcgcacg agctcggtcc attcgtcggc cggtatatgg gcattcagcg acagcagggc
     4741 gcggcgctgc gccttgtcgg cgacggggat ggagtagcca ttgtcatcga tgccgtgttt
     4801 ctgggcgtcg accagcatgg cataggcctc cggcgtcggt tcgacctcgc tccagtcgaa
     4861 gggcagctgg cgttcgaagc cctgcttgat gatcggatcg accttcacat agcagttgag
     4921 gaggtaacgg gaaacccagg catccggata ggtggtgcgc acgaagggcg aatcgatctt
     4981 gctcgcgatt gtctgggcga gatggtaggt gacgaaatcg aggttatatt ccgcctggag
     5041 aatacggatg gcggcatcca cgttggccgc cgctttgatc tgttcgaacg cggactcgaa
     5101 cttttcgcta taggtattct caatcattcg ccgttctccg ttgacgggcg tctcggacct
     5161 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggcgcg ccgcccaatt
     5221 attgaaggcc tccctaacgg ggggcctttt tttgtttctg gtctcccgct taacgatcgt
     5281 tggctgccct ttgtgcgtcc aaacggacgc acggcgctct aaagcgggtc gcgatctttc
     5341 agattcgctc ctcgcgcttt cagtctttgt tttggcgcat gtcgttatcg caaaaccgct
     5401 gcacactttt gcgcgacatg ctctgatccc cctcatctgg gggggcctat ctgagggaat
     5461 ttccgatccg gctcgcctga accattctgc tttccacgaa cttgaaaacg ctagctgtca
     5521 ccggatgtgc tttccggtct gatgagtccg tgaggacgaa acagcctcta caaataattt
     5581 tgtttaagaa ctgggaagag cgcagattaa agaggagaaa tactagatgg cttcctccga
     5641 agacgttatc aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca
     5701 cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa
     5761 actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt
     5821 ccagtacggt tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact
     5881 gtccttcccg gaaggtttca aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt
     5941 taccgttacc caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg
     6001 tggtaccaac ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc
     6061 ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct
     6121 gaaactgaaa gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa
     6181 accggttcag ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa
     6241 cgaagactac accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc
     6301 ttaataacgc tgatagtgct agtgtagatc gcgctttcca gtcgggaaac ctgtcgtgcc
     6361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgcatgc
     6421 ataaaaactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     6481 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     6541 tggacgcaca ccgtggaaac ggatgaaggc acgaacccag ttgacataag cctgttcggt
     6601 tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     6661 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt
     6721 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     6781 gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc
     6841 ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg
     6901 accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag
     6961 ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga
     7021 cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg
     7081 ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg
     7141 gagagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca
     7201 acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg
     7261 ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta
     7321 ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta
     7381 aattgtcaca acgccgca
//
