LOCUS       sequence-333084-        6840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, pSa origin, VanRAM repressor, 
            PVanCC promoter, mRFP, Gentamicin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6840)
  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 6840)
  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: pWVR5
FEATURES             Location/Qualifiers
     source          1..6840
                     /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        4957..5006
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             5051..5062
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5069..5746
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(6023..6028,6029..6045,6046..6051)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6240..6773
                     /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 ggcgacgtgc 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 ggccagtgaa acagacgaag aatccatggt caatctgcac
     4021 gaattgacca tgctgcaccc agcggtgcgc ctgcgcttgc tgctgcttca aaaacttttg
     4081 cattacgaat tgctgccagt gcatgatcac gcataatacg ttctgcacct tcggcatcac
     4141 cacagctaac tgcatccaga actgcctgat gctgacgatg tgctgccagc agatgttcat
     4201 attcggcaga caggtccatc agatccaggg ccagtgcacc ggctgctgca aacggttcaa
     4261 aaccattacg tgccagtgcg ctttcaactg caccattacc tgctgcgcta accagggtat
     4321 catgaaatgc ctgattatat gcggcataac gatccagatc ttcaccattc aggcgaccgg
     4381 ctgcaaacag tgcttcacct tctgcaatca gtacaacaaa acgtgcatgg gtttctgagg
     4441 tcataccacg ttctgccaga cgacgtgctg caaaaccttc cagaacacca cgaacttcaa
     4501 ttgcatcacg aatctgatcg ctgctaacac cacgggctgc ataaccacgt gcacccagac
     4561 gaacaaccag accttcttgt tccagtgaac gcagtgcgat acgaaccggc atacggctaa
     4621 cacccagtgc tgctgcggtc ggaatttctg caatacgttc accacttttg atttcaccgc
     4681 ttgcaatcat tttacgcagt gccatcataa cacgctgacc cggtttaata cgaggcatgt
     4741 ccatgtctat cctccttatt tacgttcgtt agtttaagcg ggcgctatca tgccataccg
     4801 cgaaaggttt tgcgccattc gatggcgcgc cgccaattat tgaaggcctc cctaacgggg
     4861 ggcctttttt tgtttctggt ctcccgctta acgatcgttg gctgattgga tccaattgac
     4921 agctagctca gtcctaggta ccattggatc caatagctgt caccggatgt gctttccggt
     4981 ctgatgagtc cgtgaggacg aaacagcctc tacaaataat tttgtttaag aactgggaag
     5041 agcgcagatt aaagaggaga aatactagat ggcttcctcc gaagacgtta tcaaagagtt
     5101 catgcgtttc aaagttcgta tggaaggttc cgttaacggt cacgagttcg aaatcgaagg
     5161 tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct aaactgaaag ttaccaaagg
     5221 tggtccgctg ccgttcgctt gggacatcct gtccccgcag ttccagtacg gttccaaagc
     5281 ttacgttaaa cacccggctg acatcccgga ctacctgaaa ctgtccttcc cggaaggttt
     5341 caaatgggaa cgtgttatga acttcgaaga cggtggtgtt gttaccgtta cccaggactc
     5401 ctccctgcaa gacggtgagt tcatctacaa agttaaactg cgtggtacca acttcccgtc
     5461 cgacggtccg gttatgcaga aaaaaaccat gggttgggaa gcttccaccg aacgtatgta
     5521 cccggaagac ggtgctctga aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg
     5581 tcactacgac gctgaagtta aaaccaccta catggctaaa aaaccggttc agctgccggg
     5641 tgcttacaaa accgacatca aactggacat cacctcccac aacgaagact acaccatcgt
     5701 tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt gcttaataac gctgatagtg
     5761 ctagtgtaga tcgcgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5821 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcat gcataaaaac tgttgtaatt
     5881 cattaagcat tctgccgaca tggaatccat cacaaacggc atgatgaacc tgaatcgcca
     5941 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggacgca caccgtggaa
     6001 acggatgaag gcacgaaccc agttgacata agcctgttcg gttcgtaaac tgtaatgcaa
     6061 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     6121 cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta tgcctcgggc
     6181 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     6241 tgttacgcag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaggtg
     6301 gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc aaatccatgc
     6361 gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac tcccaacatc
     6421 agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc gcgcttgctg
     6481 ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc aagtttgagc
     6541 agccgcgtag tgagatctat atctatgatc tcgcagtctc cggagagcac cggaggcagg
     6601 gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt ggtgcttatg
     6661 tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat acaaagttgg
     6721 gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc taacaattcg
     6781 ttcaagccga gatcggcttc ccggccgcgg agttgttcgg taaattgtca caacgccgca
//
