LOCUS       sequence-269855-        8459 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Insertion of the pHSP12 promoter and 24 PP7 stem loops in the GLT1 
            locus using HIS marker in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8459)
  AUTHORS   Wosika V, Pelet S
  TITLE     Single-particle imaging of stress-promoters induction reveals the 
            interplay between MAPK signaling, chromatin and transcription 
            factors.
  JOURNAL   Nat Commun. 2020 Jun 23;11(1):3171. doi: 10.1038/s41467-020-16943-w.
  PUBMED    32576833
REFERENCE   2  (bases 1 to 8459)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-06-23"; volume: "11(1):3171. doi"; pages: " 
            10.1038/s41467-020-16943-w"
COMMENT     Sequence Label: pVW293
FEATURES             Location/Qualifiers
     source          1..8459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        416..759
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             760..1458
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1462..1659
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4534..4553)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4535..4553)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4560..4577)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4560..4576)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4569..4591)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4714..5016
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4895..4914
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5365..6159
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5365..6159
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5419..5438)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6029..6048
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(6705..7083)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     7030..7049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7391..7979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7880..7899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8133..8150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8267..8288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8303..8308,8309..8326,8327..8333)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8341..8357
                     /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)."
     primer_bind     8346..8368
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8365..8381
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8365..8381
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8402..8420
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaattaaac cgttcaaccc tatacagcgc aaatttggtc ctaatacact tcggttttaa
       61 tgcgtcaatc cgattggctc cgataagctt ttgcacattt ttcaagtcat atgtcacgac
      121 gaacgtgatg cccgcaaaac gtaaaaaaaa gaagcaataa attgcccaaa aaaaaaatat
      181 aaactgcttt tcttagtcat tcataagttc tgatggtctt ccttatctta ttgtattttt
      241 cgtcctattg tttcatttct taccaaatta atcaattctt atatcttact tgataacaca
      301 ccaaactaat cgtctccaca tcataggaag ataggaaatt gctatctcag tcctatacta
      361 cgcagacctc gagtctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat
      421 ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga
      481 ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt
      541 tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg
      601 aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat
      661 aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct
      721 aggatacagt tctcacatca catccgaaca taaacaacca tggcagaacc agcccaaaaa
      781 aagcaaaaac aaactgttca ggagcgcaag gcgtttatct cccgtatcac taatgaaact
      841 aaaattcaaa tcgctatttc gctgaatggt ggttatattc aaataaaaga ttcgattctt
      901 cctgcaaaga aggatgacga tgtagcttcc caagctactc agtcacaggt catcgatatt
      961 cacacaggtg ttggcttttt ggatcatatg atccatgcgt tggcaaaaca ctctggttgg
     1021 tctcttattg ttgaatgtat tggtgacctg cacattgacg atcaccatac taccgaagat
     1081 tgcggtatcg cattagggca agcgttcaaa gaagcaatgg gtgctgtccg tggtgtaaaa
     1141 agattcggta ctgggttcgc accattggat gaggcgctat cacgtgccgt agtcgattta
     1201 tctagtagac catttgctgt aatcgacctt ggattgaaga gagagatgat tggtgattta
     1261 tccactgaaa tgattccaca ctttttggaa agtttcgcgg aggcggccag aattactttg
     1321 catgttgatt gtctgagagg tttcaacgat caccacagaa gtgagagtgc gttcaaggct
     1381 ttggctgttg ccataagaga agctatttct agcaatggca ccaatgacgt tccctcaacc
     1441 aaaggtgttt tgatgtgaag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     1501 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     1561 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     1621 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     1681 cgccatccag agctctgata taggacttct ctctcttttt tttttttttt gtccaggtgg
     1741 agtgcgattt gttcgttata tgcctcattc cttaattttt atcagtcaga acgttatttc
     1801 gcgctgcaag ttccttactt ttccatagag ttccacagga tgtaaagaaa aataatgata
     1861 aaaaaaagca gaacatatta cccttctttt ggagcgtact tccatctgtt acccgctcat
     1921 ccaaatgcac attctgttac cctcttgttc ggtatttttc ggagttctcc ttttctcctt
     1981 cttttcctct tgatacacag aaaagtggga gaaggtaggg ggagtcagag aatgaagaaa
     2041 aaaaggggac gcgcaaatcc aagtgaaaat ctccgggagc gggcggatcc cactaacggc
     2101 ccagccgaaa atggaaaaaa agggtcggtg atgtgtgggt gccagctggc ggtagcaatg
     2161 acgacgtgtt gacgggccct tggctcttgg gacaaggact agaagccaaa agccagaggc
     2221 ggtaaaaata gcaagactag aatattgctg gcatctgtta aggggatatg ttgcaacttg
     2281 cagggggcgg cacaaaataa catagaaacg tagtaaagag gggaaaagga aaaggaaaag
     2341 gaaaaggaag gaaaaaaacc cattgacgta gaaattgaaa gaaggaaagg tatacgcaag
     2401 cattaataca acccacaaac acagaccaga agcactctag acggagagta actagatcta
     2461 cagcccctgg aaaatcgttt ggtcaacttt gaggttccgg tcgtccccct cttgatctga
     2521 aaggtctttc tctaaatcta tattaaaacg tataaatagg acggtgaatt gcgttctact
     2581 tcctcaattg cgtttgatct tatttaatct ctctctaata tatagaaaaa aaaaccatct
     2641 gattattcga taatctcaaa caaacaactc aaaacaaaaa aaactaaata caacaccgcg
     2701 gggatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca
     2761 ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca
     2821 ttagatccta aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc
     2881 aggtcgactc tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc
     2941 attagatcct aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg
     3001 caggtcgact ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt
     3061 cattagatcc taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct
     3121 gcaggtcgac tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt
     3181 tcattagatc ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc
     3241 tgcaggtcga ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg
     3301 ttcattagat cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc
     3361 ctgcaggtcg actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg
     3421 gttcattaga tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc
     3481 cctgcaggtc gactctagaa accagcagag catatgggct cgctggctgc agtattcccg
     3541 ggttcattag atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct
     3601 ccctgcaggt cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc
     3661 gggttcatta gatcctaagg tacctaattg cctagaaagg agcagacgat atggcgtcgc
     3721 tccctgcagg tcgactctag aaaccagcag agcatatggg ctcgctggct gcagtattcc
     3781 cgggttcatt agatcctaag gtacctaatt gcctagaaag gagcagacga tatggcgtcg
     3841 ctccctgcag gtcgactcta gaaaccagca gagcatatgg gctcgctggc tgcagtattc
     3901 ccgggttcat tagatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc
     3961 gctccctgca ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt
     4021 cccgggttca ttagatccta aggtacctaa ttgcctagaa aggagcagac gatatggcgt
     4081 cgctccctgc aggtcgactc tagaaaccag cagagcatat gggctcgctg gctgcagtat
     4141 tcccgggttc attagatcca tgtcccagga tagaattttg ttagatagat ctatgccagt
     4201 gttgaaatca gacaatttcg atccattgga agaagcttac gaaggtggga caattcaaaa
     4261 ctataacgat gaacaccatc ttcataaatc ttgggcaaat gtgattccgg acaaacgagg
     4321 actttacgac cctgattatg aacatgacgc ttgtggtgtc ggtttcgtag caaataagca
     4381 tggtgaacag tctcacaaga ttgttactga cgctagatat cttttagtga atatgacaca
     4441 tcgtggtgcc gtctcatctg atgggaacgg tgacggtgcc ggtattctgc taggtatccc
     4501 aagggcgaat tcgcggccgc taaattcaat tcgccctata gtgagtcgta ttacaattca
     4561 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4621 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4681 ccttcccaac agttgcgcag cctatacgta cggcagttta aggtttacac ctataaaaga
     4741 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gccggggcga
     4801 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     4861 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     4921 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     4981 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggcatgag attatcaaaa
     5041 aggatcttca cctagatcct tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc
     5101 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
     5161 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
     5221 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
     5281 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag ctctgatcaa
     5341 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5401 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5461 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5521 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     5581 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5641 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5701 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5761 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5821 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5881 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5941 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6001 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6061 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6121 cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttattaacgc ttacaatttc
     6181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat caggtggcac
     6241 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6301 gtatccgctc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6361 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6421 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6481 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6541 gataccgcga gaccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6601 aggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6661 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6721 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6781 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6841 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6901 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6961 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7021 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7081 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7141 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7201 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7261 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7321 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7381 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7441 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7501 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7561 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7621 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7681 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7741 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7801 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7861 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7921 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7981 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8041 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8101 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     8161 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     8221 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     8281 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     8341 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
     8401 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc ccttatata
//
