LOCUS       sequence-314925-        6848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, incW (pSa) origin, LuxR 
            activator, PLuxB promoter, mRFP, Gentamicin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6848)
  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 6848)
  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: pWLxR5
FEATURES             Location/Qualifiers
     source          1..6848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    28..48
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      49..106
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(130..147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             951..1922
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein of plasmid pSa"
                     /label=RepA
                     /translation="MPKNNKAPGHRINEIIKTSLALEMEDAREAGLVGYMARCLVQATM
                     PHTDPKTSYFERTNGIVTLSIMGKPSIGLPYGSMPRTLLAWICTEAVRTKDPVLNLGRS
                     QSEFLQRLGMHTDGRYTATLRNQAQRLFSSMISLAGEQGNDFGIENVVIAKRAFLFWNP
                     KRPEDRALWDSTLTLTGDFFEEVTRSPVPIRIDYLHALRQSPLAMDIYTWLTYRVFLLR
                     AKGRPFVQIPWVALQAQFGSSYGSRARNSPELDDKARERAERAALASFKYNFKKRLREV
                     LIVYPEASDCIEDDGECLRIKSTRLHVTRAPGKGARIGPPPT"
     rep_origin      1959..2394
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     primer_bind     3970..3987
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3971..3987
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        4965..5014
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             5059..5070
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5077..5754
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(6031..6036,6037..6053,6054..6059)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6248..6781
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ggtggcactt ttcggggaga catcgcttac tagtagcggc cgctgcagtc cggcaaaaaa
       61 gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat
      121 gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      181 agctcactca aaggcggtaa tcaaaccaac gttttatcta taccgctaca gggtatttaa
      241 ttcctattta atctgcgcta gaatgaggca tgtttaaccg aatctgacgt tttccctgca
      301 aatgccaaaa tactatgcct atctccgggt ttcgcgtgac ggccaagacc cggaaaacca
      361 aaaatacggt ttgctcgaat acgcgaacgc caaaggcttc gcgccgctac agatcgagga
      421 agaaattgcc agcagagcaa aggactggcg caagcgcaag ctcggagcaa tcatcgaaaa
      481 ggccgagcgt agcgacgtgc tactgacgcc ggagattacg cgcattgccg gttccgccct
      541 cgccgccttg gaaattctca aagcggcgag cgagcgcggc ctaatcgtcc atgtgaccaa
      601 acagaagatc atcatggacg gcagcctaca aagcgacatc atggcaaccg tgcttggctt
      661 ggctgcacag atcgagcggc atttcattca ggcacgtacc accgaggcgc tacaagtcgc
      721 cagagagcgc ggcaagacgc tcgggcgacc caagggcagc aaatcgagcg ccttgaagct
      781 ggacagccgt attgatgaag tacaggcata cgtgaacctt ggcttgccgc aaagtcgcgc
      841 agccgagttg ttaggcgtca gccctcacac cttgcgcctg ttcatcaaac gccggaacat
      901 caaacccaca aacactagac caaccatcac catgccgggg agggaacaac atgcctaaga
      961 acaacaaagc ccccggccat cgtatcaacg agatcatcaa gacgagcctc gcgctcgaaa
     1021 tggaggatgc ccgcgaagct ggcttagtcg gctacatggc ccgttgcctt gtgcaagcga
     1081 ccatgcccca caccgacccc aagaccagct actttgagcg caccaatggc atcgtcacct
     1141 tgtcgatcat gggcaagccg agcatcggcc tgccctacgg ttctatgccg cgcaccttgc
     1201 ttgcttggat atgcaccgag gccgtgcgaa cgaaagaccc cgtgttgaac cttggccggt
     1261 cgcaatcgga atttctacaa aggctcggaa tgcacaccga tggccgttac acggccaccc
     1321 ttcgcaatca ggcgcaacgc ctgttttcat ccatgatttc gcttgccggc gagcaaggca
     1381 atgacttcgg cattgagaac gtcgtcattg ccaagcgcgc ttttctattc tggaatccca
     1441 agcggccaga agatcgggcg ctatgggata gcaccctcac cctcacaggc gatttcttcg
     1501 aggaagtcac ccgctcaccg gttcctatcc gaatcgacta cctgcatgcc ttgcggcagt
     1561 ctccgcttgc gatggacatt tacacgtggc tgacctatcg cgtgttcctg ttgcgggcca
     1621 agggccgccc cttcgtgcaa atcccttggg tcgccctgca agcgcaattc ggctcatcct
     1681 atggcagccg cgcacgcaac tcgcccgaac tggacgataa ggcccgagag cgggcagagc
     1741 gggcagcact cgccagcttc aaatacaact tcaaaaagcg cctacgcgaa gtgttgattg
     1801 tctatcccga ggcaagcgac tgcatcgaag atgacggcga atgcctgcgc atcaaatcca
     1861 cacgcctgca tgtcacccgc gcacccggca agggcgctcg catcggcccc cctccgactt
     1921 gaccaggcca acgctacgct tggcttggtc aagccttccc atccaacagc ccgccgtcga
     1981 gcgggctttt ttatccccgg aagcctgtgg atagagggta gttatccacg tgaaaccgct
     2041 aatgccccgc aaagccttga ttcacggggc tttccggccc gctccaaaaa ctatccacgt
     2101 gaaatcgcta atcagggtac gtgaaatcgc taatcggagt acgtgaaatc gctaataagg
     2161 tcacgtgaaa tcgctaatca aaaaggcacg tgagaacgct aatagccctt tcagatcaac
     2221 agcttgcaaa cacccctcgc tccggcaagt agttacagca agtagtatgt tcaattagct
     2281 tttcaattat gaatatatat atcaattatt ggtcgccctt ggcttgtgga caatgcgcta
     2341 cgcgcaccgg ctccgcccgt ggacaaccgc aagcggttgc ccaccgtcga gcgccagcgc
     2401 ctttgcccac aacccggcgg ccgcaacaga tcgttttata aatttttttt tttgaaaaag
     2461 aaaaagcccg aaaggcggca acctctcggg cttctggatt tccgatcaac gcaggagtcg
     2521 ttcggaaagt agctgttcca gaattatagg cgcagagaca ccagattcca agatggctct
     2581 gttaaattgt tgtagtatgt agtatcatac aacatactac agtacagagg cccgcaagaa
     2641 tggcaatcac taaacaagac atttggcgag cagccgacga actggacgcc gaaggcatcc
     2701 ggcccacttt ggccgccgtg cgcaagaaac tcggaagcgg tagcttcaca accatttccg
     2761 atgcaatggc tgaatggaaa aaccgcaaga ccgccaccct gccctcatca gacccattgc
     2821 cggttgcagt caacgagcat cttgccgagc ttggcaatgc gctatgggct atcgccctgg
     2881 cgcacgccaa cgcccggttt gacgaagatc ggaaacagat cgaggccgac aaagcggcca
     2941 tcagccagca gcttgccgaa gcaatcgaac tagccgacac cttcacccgc gaaaacgacc
     3001 agctccgcga acgagtgaat cagctcgaac ctatggaacg cgagcgcgac aagctggccg
     3061 accagcttgc cgaagtgaaa cgccgcagcg gcgaagaact aaaccgctgc atggaaaagc
     3121 tcacccaacg cgataacgag gctatcgagg cccgcaaaca ggccaaggag gccatcgagc
     3181 gcgccgccag tctgcaaggt caggtggaag ccctcaaaga gcaggtcgcc aatctcacag
     3241 ccgtcttgaa aacaggaggc aaacaatgaa aagcgccctt gccgcccttc gcgcggtcgc
     3301 ggccgctgtc gtcctaatcg tcagtgtgcc cgcttgggcc gacttccggg gtgaagtcgt
     3361 ccgaatcctt gacggtgaca ctatcgacgt tttggtgaac cgtcagacca tccgcgtgag
     3421 attggccgat attgacgcac cggaaagcgg ccaagccttc ggctcccgtg ctcgccaacg
     3481 gctcgccgac ttgacctttc gccaagaggt tcaagtgacc gaaaaagagg ttgatcggta
     3541 tggccgcact cttggggtcg tttacgcgcc gttgcaatac cccggcggcc aaacacaact
     3601 caccaacatc aatgcgatca tggttcaaga aggcatggcc tgggcttacc gttattacgg
     3661 caaaccaacc gacgcgcaga tgtacgagta tgaaaaagag gcccgccgcc aacggctcgg
     3721 cctttggtca gacccgaatg ctcaggagcc ttggaaatgg cgtcgcgcct cgaaaaatgc
     3781 cacgaactga caccgggcac gccccttgtt cgacgcgccg caggcacgtc gaatttaccg
     3841 ccgggacgcc cctcgtcccg acacttccag atcgccatag cgcacagcgc cgattatcaa
     3901 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagtg
     3961 tcacgacgtt gtaaaacgac ggccagtgat caatttttaa agtatgggca atcaattgct
     4021 cctgttaaaa ttgctttaga aatactttgg cagcggtttg ttgtattgag tttcatttgc
     4081 gcattggtta aatggaaagt gacagtacgc tcactgcaac ctaatatttt tgaaatatcc
     4141 caagagcttt ttccttcgca tgcccacgct aaacattctt tttctctttt ggttaaatcg
     4201 ttgtttgatt tattatttgc tatatttatt tttcgataat tatcaactag agaaggaaca
     4261 attaatggta tgttcataca cgcatgtaaa aataaactat ctatatagtt gtctttttct
     4321 gaatgtgcaa aactaagcat tccgaagcca ttgttagccg tatgaatagg gaaactaaac
     4381 ccagtgataa gacctgatgt tttcgcttct ttaattacat ttggagattt tttatttaca
     4441 gcattgtttt caaatatatt ccaattaatt ggtgaatgat tggagttaga ataatctact
     4501 ataggatcat attttatcaa attagcgtca tcataatatt gcctccattt tttagggtaa
     4561 ttatctagga ttgaaatatc agatttaacc atagaatgag gataaatgat cgcgagtaaa
     4621 taatattcac aatgtaccat tttagtcata tcagataagc attgattaat atcattattg
     4681 cttctacaag ctttaatttt attaattatt ctgtatgtgt cgtcggcatt tatgtttttc
     4741 attctgcccc tccgaaaggc ttaagtcctc gcgtagtacc tgggcgctat catgccatac
     4801 cgcgaaaggt tttgcgccat tcgatggcgc gccgcccaat tattgaaggc ctccctaacg
     4861 gggggccttt ttttgtttct ggtctcccgc ttaacgatcg ttggctgacc tgtaggatcg
     4921 tacaggttta cgcaagaaaa tggtttgtta cagtcgaata aaagctgtca ccggatgtgc
     4981 tttccggtct gatgagtccg tgaggacgaa acagcctcta caaataattt tgtttaagaa
     5041 ctgggaagag cgcagattaa agaggagaaa tactagatgg cttcctccga agacgttatc
     5101 aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa
     5161 atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt
     5221 accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt
     5281 tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg
     5341 gaaggtttca aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt taccgttacc
     5401 caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggtaccaac
     5461 ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa
     5521 cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa
     5581 gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag
     5641 ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaagactac
     5701 accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc ttaataacgc
     5761 tgatagtgct agtgtagatc gcgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5821 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgcatgc ataaaaactg
     5881 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     5941 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggacgcaca
     6001 ccgtggaaac ggatgaaggc acgaacccag ttgacataag cctgttcggt tcgtaaactg
     6061 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     6121 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt acagtctatg
     6181 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     6241 agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     6301 gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa
     6361 atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag ccacctactc
     6421 ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc
     6481 gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccaa
     6541 gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg gagagcaccg
     6601 gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg
     6661 tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac
     6721 aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta
     6781 acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta aattgtcaca
     6841 acgccgca
//
