LOCUS       sequence-314926-        7203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, incW (pSa) 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 7203)
  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 7203)
  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: pWLlR5
FEATURES             Location/Qualifiers
     source          1..7203
                     /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"
     CDS             complement(4038..5120)
                     /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     5082..5101
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5121..5198)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      complement(5262..5305)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5308..5329
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    5332..5348
                     /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    5355..5371
                     /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             5414..5425
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5432..6109
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(6386..6391,6392..6408,6409..6414)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6603..7136
                     /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 ggccagtgag atcccggtgc ctaatgagtg agctaactta
     4021 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4081 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     4141 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     4201 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     4261 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     4321 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     4381 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     4441 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     4501 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     4561 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4621 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4681 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     4741 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     4801 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     4861 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     4921 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     4981 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     5041 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     5101 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     5161 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     5221 atgcgactcc tgcattagga aatggcgtat cacgaggcag aatttcagat aaaaaaaatc
     5281 cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagaga attgtgagcg
     5341 gataacaatt gacattgtga gcggataaca agatactgag cacaccctgt aggaactggg
     5401 aagagcgcag attaaagagg agaaatacta gatggcttcc tccgaagacg ttatcaaaga
     5461 gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga
     5521 aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga aagttaccaa
     5581 aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt acggttccaa
     5641 agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct tcccggaagg
     5701 tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg ttacccagga
     5761 ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta ccaacttccc
     5821 gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca ccgaacgtat
     5881 gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg
     5941 tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc
     6001 gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag actacaccat
     6061 cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat aacgctgata
     6121 gtgctagtgt agatcgcgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6181 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg catgcataaa aactgttgta
     6241 attcattaag cattctgccg acatggaatc catcacaaac ggcatgatga acctgaatcg
     6301 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggac gcacaccgtg
     6361 gaaacggatg aaggcacgaa cccagttgac ataagcctgt tcggttcgta aactgtaatg
     6421 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     6481 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttgtacagt ctatgcctcg
     6541 ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa
     6601 cgatgttacg cagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttag
     6661 gtggctcaag tatgggcatc attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca
     6721 tgcgggctgc tcttgatctt ttcggtcgtg agttcggaga cgtagccacc tactcccaac
     6781 atcagccgga ctccgattac ctcgggaact tgctccgtag taagacattc atcgcgcttg
     6841 ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc ttacgttctg cccaagtttg
     6901 agcagccgcg tagtgagatc tatatctatg atctcgcagt ctccggagag caccggaggc
     6961 agggcattgc caccgcgctc atcaatctcc tcaagcatga ggccaacgcg cttggtgctt
     7021 atgtgatcta cgtgcaagca gattacggtg acgatcccgc agtggctctc tatacaaagt
     7081 tgggcatacg ggaagaagtg atgcactttg atatcgaccc aagtaccgcc acctaacaat
     7141 tcgttcaagc cgagatcggc ttcccggccg cggagttgtt cggtaaattg tcacaacgcc
     7201 gca
//
