LOCUS       sequence-494926-        8346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insertion of the pGRE2 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 8346)
  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 8346)
  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: "Nat 
            Commun."; date: "2020-06-23"; volume: "11(1):3171. doi"; pages: " 
            10.1038/s41467-020-16943-w"
COMMENT     Sequence Label: pVW294
FEATURES             Location/Qualifiers
     source          1..8346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        370..713
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             714..1412
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1416..1613
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     2650..2666
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2650..2666
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4373..4392)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4374..4392)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4399..4416)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4399..4415)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4408..4430)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4553..4855
                     /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     4734..4753
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5204..5998
                     /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             5204..5998
                     /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(5258..5277)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5868..5887
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6248..7039,7040..7108))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6871..6890
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7232..7820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7721..7740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7974..7991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8108..8129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8144..8149,8150..8167,8168..8174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8182..8198
                     /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     8187..8209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8206..8222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8206..8222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8243..8261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cacttcggtt ttaatgcgtc aatccgattg gctccgataa gcttttgcac atttttcaag
       61 tcatatgtca cgacgaacgt gatgcccgca aaacgtaaaa aaaagaagca ataaattgcc
      121 caaaaaaaaa atataaactg cttttcttag tcattcataa gttctgatgg tcttccttat
      181 cttattgtat ttttcgtcct attgtttcat ttcttaccaa attaatcaat tcttatatct
      241 tacttgataa cacaccaaac taatcgtctc cacatcatag gaagatagga aattgctatc
      301 tcagtcctat actacgcaga cctcgagtct gtttagcttg cctcgtcccc gccgggtcac
      361 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
      421 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
      481 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
      541 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
      601 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
      661 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatggcag
      721 aaccagccca aaaaaagcaa aaacaaactg ttcaggagcg caaggcgttt atctcccgta
      781 tcactaatga aactaaaatt caaatcgcta tttcgctgaa tggtggttat attcaaataa
      841 aagattcgat tcttcctgca aagaaggatg acgatgtagc ttcccaagct actcagtcac
      901 aggtcatcga tattcacaca ggtgttggct ttttggatca tatgatccat gcgttggcaa
      961 aacactctgg ttggtctctt attgttgaat gtattggtga cctgcacatt gacgatcacc
     1021 atactaccga agattgcggt atcgcattag ggcaagcgtt caaagaagca atgggtgctg
     1081 tccgtggtgt aaaaagattc ggtactgggt tcgcaccatt ggatgaggcg ctatcacgtg
     1141 ccgtagtcga tttatctagt agaccatttg ctgtaatcga ccttggattg aagagagaga
     1201 tgattggtga tttatccact gaaatgattc cacacttttt ggaaagtttc gcggaggcgg
     1261 ccagaattac tttgcatgtt gattgtctga gaggtttcaa cgatcaccac agaagtgaga
     1321 gtgcgttcaa ggctttggct gttgccataa gagaagctat ttctagcaat ggcaccaatg
     1381 acgttccctc aaccaaaggt gttttgatgt gaagtactga caataaaaag attcttgttt
     1441 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     1501 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     1561 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     1621 ttcgatacta acgccgccat ccagagctca agcatttctg caaaaccatg taaatatttg
     1681 attattagcg ggtaaatatg taatagttat gcactttaat tcttttgttc atgtagaaga
     1741 caatacaacg catttcttgg ctaaccatct tacagccttg agggtaaagg gcgggcaaac
     1801 gaacaaaagt acaaaatttc catccttgtt tgataatcat ttcctcggac cagataagaa
     1861 tgagcagccg aaataaacgg taacgtgatt aaaaccgcta gaagagtgtt tttgtaagga
     1921 tgaaaacctc agtgagaaag ttgacaatat aaacattatc agcttttctc gccctctttt
     1981 gcacttcctg caacattact tgttctgcag cttgtttctc accaataaca gaacatatcc
     2041 agtataatgc atacaccata ttcacaccct atttcatatc attaacctct tcatttttag
     2101 atgccagcga ttactttgac ttctaaccaa gaccgcgaaa aaaggttacc gcgtgaacta
     2161 tttcatattt gcgattttaa gtacaataaa tattatcatt attatattat gtttgcatgt
     2221 aggttctaca aatacattgt tgtacgctat agtttccttc caaaactaaa aagagttcgt
     2281 aacaaaataa tcttcaatat ttcatagtac cttattgata tcgatcctgc gacaccttct
     2341 catttcaaca attggccctc acctcttttg tgcaaaaaac gtcgccattg ataaaataag
     2401 caagaagcat ataattggaa tgtccattac gtaaatgaaa aaaaatcatt ttacatatta
     2461 cgtaatggaa tacggaattt tctcgcggaa gtagatcttc cgtggaaaaa aaggaaaaag
     2521 tccgatcaat attgaaaaag gcatccttag tttctcaact atataaggag gaaaagtcca
     2581 cctgtgtagc gttgatataa cgtattcgat tttcaaacga aaagatagca gtatcacacg
     2641 cccttaaatt ctagaactag tggatcctaa ggtacctaat tgcctagaaa ggagcagacg
     2701 atatggcgtc gctccctgca ggtcgactct agaaaccagc agagcatatg ggctcgctgg
     2761 ctgcagtatt cccgggttca ttagatccta aggtacctaa ttgcctagaa aggagcagac
     2821 gatatggcgt cgctccctgc aggtcgactc tagaaaccag cagagcatat gggctcgctg
     2881 gctgcagtat tcccgggttc attagatcct aaggtaccta attgcctaga aaggagcaga
     2941 cgatatggcg tcgctccctg caggtcgact ctagaaacca gcagagcata tgggctcgct
     3001 ggctgcagta ttcccgggtt cattagatcc taaggtacct aattgcctag aaaggagcag
     3061 acgatatggc gtcgctccct gcaggtcgac tctagaaacc agcagagcat atgggctcgc
     3121 tggctgcagt attcccgggt tcattagatc ctaaggtacc taattgccta gaaaggagca
     3181 gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac cagcagagca tatgggctcg
     3241 ctggctgcag tattcccggg ttcattagat cctaaggtac ctaattgcct agaaaggagc
     3301 agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa ccagcagagc atatgggctc
     3361 gctggctgca gtattcccgg gttcattaga tcctaaggta cctaattgcc tagaaaggag
     3421 cagacgatat ggcgtcgctc cctgcaggtc gactctagaa accagcagag catatgggct
     3481 cgctggctgc agtattcccg ggttcattag atcctaaggt acctaattgc ctagaaagga
     3541 gcagacgata tggcgtcgct ccctgcaggt cgactctaga aaccagcaga gcatatgggc
     3601 tcgctggctg cagtattccc gggttcatta gatcctaagg tacctaattg cctagaaagg
     3661 agcagacgat atggcgtcgc tccctgcagg tcgactctag aaaccagcag agcatatggg
     3721 ctcgctggct gcagtattcc cgggttcatt agatcctaag gtacctaatt gcctagaaag
     3781 gagcagacga tatggcgtcg ctccctgcag gtcgactcta gaaaccagca gagcatatgg
     3841 gctcgctggc tgcagtattc ccgggttcat tagatcctaa ggtacctaat tgcctagaaa
     3901 ggagcagacg atatggcgtc gctccctgca ggtcgactct agaaaccagc agagcatatg
     3961 ggctcgctgg ctgcagtatt cccgggttca ttagatccat gtcccaggat agaattttgt
     4021 tagatagatc tatgccagtg ttgaaatcag acaatttcga tccattggaa gaagcttacg
     4081 aaggtgggac aattcaaaac tataacgatg aacaccatct tcataaatct tgggcaaatg
     4141 tgattccgga caaacgagga ctttacgacc ctgattatga acatgacgct tgtggtgtcg
     4201 gtttcgtagc aaataagcat ggtgaacagt ctcacaagat tgttactgac gctagatatc
     4261 ttttagtgaa tatgacacat cgtggtgccg tctcatctga tgggaacggt gacggtgccg
     4321 gtattctgct aggtatccca agggcgaatt cgcggccgct aaattcaatt cgccctatag
     4381 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     4441 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     4501 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctatacgtac ggcagtttaa
     4561 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     4621 tattgacacg ccggggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     4681 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     4741 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     4801 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     4861 aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga aagccagtcc
     4921 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
     4981 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
     5041 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
     5101 tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag
     5161 gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5221 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5281 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5341 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     5401 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5461 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5521 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5581 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5641 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5701 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     5761 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5821 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5881 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5941 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat
     6001 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6061 acaccgcatc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6121 tctaaataca ttcaaatatg tatccgctca tgagattatc aaaaaggatc ttcacctaga
     6181 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6241 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6301 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6361 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6421 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6481 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6541 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6601 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6661 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6721 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6781 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6841 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6901 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6961 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7021 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7081 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7141 atcagggtta ttgtctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     8221 accatgatta cgccaagctc agaattaacc ctcactaaag ggactagtcc tgcaggttta
     8281 aacgaattcg cccttatata ttaattaaac cgttcaaccc tatacagcgc aaatttggtc
     8341 ctaata
//
