LOCUS       sequence-311913-        7324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RK2 and pUC origin, AraC 
            activator/repressor, PBAD promoter, mRFP, Gentamicin Resistance 
            Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7324)
  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 7324)
  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: pKAR5
FEATURES             Location/Qualifiers
     source          1..7324
                     /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     4350..4366
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4421..5299)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     5375..5394
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        5460..5509
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             5554..5565
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5572..6249
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(6526..6531,6532..6548,6549..6554)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6724..7257
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     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 gccagtgaaa ataggggttc
     4381 cgcgcacatt tccccgaaaa gtgccacctg catcgattta ttatgacaac ttgacggcta
     4441 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     4501 ttttaaatac ccgcgagaag tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     4561 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     4621 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     4681 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     4741 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     4801 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     4861 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     4921 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     4981 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     5041 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     5101 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     5161 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     5221 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     5281 cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc
     5341 cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga
     5401 ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgagc
     5461 tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg acgaaacagc ctctacaaat
     5521 aattttgttt aagaactggg aagagcgcag attaaagagg agaaatacta gatggcttcc
     5581 tccgaagacg ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac
     5641 ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc
     5701 gctaaactga aagttaccaa aggtggtccg ctgccgttcg cttgggacat cctgtccccg
     5761 cagttccagt acggttccaa agcttacgtt aaacacccgg ctgacatccc ggactacctg
     5821 aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga agacggtggt
     5881 gttgttaccg ttacccagga ctcctccctg caagacggtg agttcatcta caaagttaaa
     5941 ctgcgtggta ccaacttccc gtccgacggt ccggttatgc agaaaaaaac catgggttgg
     6001 gaagcttcca ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg
     6061 cgtctgaaac tgaaagacgg tggtcactac gacgctgaag ttaaaaccac ctacatggct
     6121 aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga catcacctcc
     6181 cacaacgaag actacaccat cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc
     6241 ggtgcttaat aacgctgata gtgctagtgt agatcgcgct ttccagtcgg gaaacctgtc
     6301 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6361 catgcataaa aactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     6421 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     6481 gcccatggac gcacaccgtg gaaacggatg aaggcacgaa cccagttgac ataagcctgt
     6541 tcggttcgta aactgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
     6601 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     6661 tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     6721 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
     6781 ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg gccctgacca agtcaaatcc
     6841 atgcgggctg ctcttgatct tttcggtcgt gagttcggag acgtagccac ctactcccaa
     6901 catcagccgg actccgatta cctcgggaac ttgctccgta gtaagacatt catcgcgctt
     6961 gctgccttcg accaagaagc ggttgttggc gctctcgcgg cttacgttct gcccaagttt
     7021 gagcagccgc gtagtgagat ctatatctat gatctcgcag tctccggaga gcaccggagg
     7081 cagggcattg ccaccgcgct catcaatctc ctcaagcatg aggccaacgc gcttggtgct
     7141 tatgtgatct acgtgcaagc agattacggt gacgatcccg cagtggctct ctatacaaag
     7201 ttgggcatac gggaagaagt gatgcacttt gatatcgacc caagtaccgc cacctaacaa
     7261 ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt tcggtaaatt gtcacaacgc
     7321 cgca
//
