LOCUS       sequence-311926-        7591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RK2 and pUC origin, LacI 
            repressor, PLlacO-1 promoter, mRFP, Gentamicin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7591)
  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 7591)
  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: pKLlR5
FEATURES             Location/Qualifiers
     source          1..7591
                     /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(4417..5499)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5461..5480
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5500..5577)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      complement(5641..5684)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5687..5708
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    5711..5727
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    5734..5750
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5802..5813
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5820..6497
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(6774..6779,6780..6796,6797..6802)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6991..7524
                     /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 gccagtgaga tcccggtgcc
     4381 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4441 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4501 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     4561 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     4621 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     4681 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     4741 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     4801 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     4861 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     4921 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     4981 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5041 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5101 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5161 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5221 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     5281 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     5341 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     5401 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     5461 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     5521 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     5581 gatctcgacg ctctccctta tgcgactcct gcattaggaa atggcgtatc acgaggcaga
     5641 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc
     5701 ttctagagaa ttgtgagcgg ataacaattg acattgtgag cggataacaa gatactgagc
     5761 acaccctgta ggaactggga agagcgcaac acatactaga gaaagaggag aaatactaga
     5821 tggcttcctc cgaagacgtt atcaaagagt tcatgcgttt caaagttcgt atggaaggtt
     5881 ccgttaacgg tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta
     5941 cccagaccgc taaactgaaa gttaccaaag gtggtccgct gccgttcgct tgggacatcc
     6001 tgtccccgca gttccagtac ggttccaaag cttacgttaa acacccggct gacatcccgg
     6061 actacctgaa actgtccttc ccggaaggtt tcaaatggga acgtgttatg aacttcgaag
     6121 acggtggtgt tgttaccgtt acccaggact cctccctgca agacggtgag ttcatctaca
     6181 aagttaaact gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca
     6241 tgggttggga agcttccacc gaacgtatgt acccggaaga cggtgctctg aaaggtgaaa
     6301 tcaaaatgcg tctgaaactg aaagacggtg gtcactacga cgctgaagtt aaaaccacct
     6361 acatggctaa aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc aaactggaca
     6421 tcacctccca caacgaagac tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc
     6481 actccaccgg tgcttaataa cgctgatagt gctagtgtag atcgcgcttt ccagtcggga
     6541 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6601 attgggcgca tgcataaaaa ctgttgtaat tcattaagca ttctgccgac atggaagcca
     6661 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     6721 taatatttgc ccatggacgc acaccgtgga aacggatgaa ggcacgaacc cagttgacat
     6781 aagcctgttc ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     6841 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     6901 actgtttttt tgtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     6961 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagcaacga tgttacgcag
     7021 cagggcagtc gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt
     7081 aggctcggcc ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag
     7141 ttcggagacg tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg
     7201 ctccgtagta agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct
     7261 ctcgcggctt acgttctgcc caagtttgag cagccgcgta gtgagatcta tatctatgat
     7321 ctcgcagtct ccggagagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc
     7381 aagcatgagg ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac
     7441 gatcccgcag tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat
     7501 atcgacccaa gtaccgccac ctaacaattc gttcaagccg agatcggctt cccggccgcg
     7561 gagttgttcg gtaaattgtc acaacgccgc a
//
