LOCUS       sequence-494935-        8020 bp    DNA     circular SYN 09-DEC-2025
DEFINITION  Insertion of the pALD3 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 8020)
  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 8020)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 9, 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: pVW295
FEATURES             Location/Qualifiers
     source          1..8020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        408..751
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             752..1450
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1454..1651
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     2362..2378
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2362..2378
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4085..4104)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4086..4104)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4111..4128)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4111..4127)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4120..4142)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4265..4567
                     /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     4446..4465
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             4916..5710
                     /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             4916..5710
                     /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(4970..4989)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5580..5599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(5960..6751,6752..6820))
                     /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     6583..6602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6944..7532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7433..7452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7686..7703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7820..7841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7856..7861,7862..7879,7880..7886)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7894..7910
                     /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     7899..7921
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7918..7934
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7918..7934
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7955..7973
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 accgttcaac cctatacagc gcaaatttgg tcctaataca cttcggtttt aatgcgtcaa
       61 tccgattggc tccgataagc ttttgcacat ttttcaagtc atatgtcacg acgaacgtga
      121 tgcccgcaaa acgtaaaaaa aagaagcaat aaattgccca aaaaaaaaat ataaactgct
      181 tttcttagtc attcataagt tctgatggtc ttccttatct tattgtattt ttcgtcctat
      241 tgtttcattt cttaccaaat taatcaattc ttatatctta cttgataaca caccaaacta
      301 atcgtctcca catcatagga agataggaaa ttgctatctc agtcctatac tacgcagacc
      361 tcgagtctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
      421 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
      481 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
      541 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
      601 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
      661 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
      721 gttctcacat cacatccgaa cataaacaac catggcagaa ccagcccaaa aaaagcaaaa
      781 acaaactgtt caggagcgca aggcgtttat ctcccgtatc actaatgaaa ctaaaattca
      841 aatcgctatt tcgctgaatg gtggttatat tcaaataaaa gattcgattc ttcctgcaaa
      901 gaaggatgac gatgtagctt cccaagctac tcagtcacag gtcatcgata ttcacacagg
      961 tgttggcttt ttggatcata tgatccatgc gttggcaaaa cactctggtt ggtctcttat
     1021 tgttgaatgt attggtgacc tgcacattga cgatcaccat actaccgaag attgcggtat
     1081 cgcattaggg caagcgttca aagaagcaat gggtgctgtc cgtggtgtaa aaagattcgg
     1141 tactgggttc gcaccattgg atgaggcgct atcacgtgcc gtagtcgatt tatctagtag
     1201 accatttgct gtaatcgacc ttggattgaa gagagagatg attggtgatt tatccactga
     1261 aatgattcca cactttttgg aaagtttcgc ggaggcggcc agaattactt tgcatgttga
     1321 ttgtctgaga ggtttcaacg atcaccacag aagtgagagt gcgttcaagg ctttggctgt
     1381 tgccataaga gaagctattt ctagcaatgg caccaatgac gttccctcaa ccaaaggtgt
     1441 tttgatgtga agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     1501 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     1561 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     1621 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     1681 agagctctta tgtaagtgcg taagtaccta aatagcttag cctgacgacg gatgaatatg
     1741 ttgacagtct agcaaacagt agtagtccac tactcgatat aaaactgtaa gcaccattat
     1801 ttcagcatgc ggaacaattg ttgttactag gttctagaaa tcccttaaga tggtaaaaaa
     1861 atagagctat aaactcagtg agatttattt ttgtgtcaag aaattatgct tgcgtgttag
     1921 ttttctctat tcccaccatt acgtagatct cccgtctttt ttttttcttt ttctgcatat
     1981 gacgtctgtt cttccttccc cccctggagt acaaggaaag tatcactatt agaactctac
     2041 agtacctttc ttccacgccc tttagtgcat ttagggattt tcattactgc taatccaaga
     2101 cttgaaccgg attgttgatc agcccccttc ttaagctgat tgcgaaactc cctaagggtt
     2161 aaggaattag taagtttagg gaggagacag ggagattgtt caaacgaata gcgtagattt
     2221 aactgatcat acctcagaaa ctaattgtat aaaatggcaa atctggatgg gaataaattt
     2281 gactcatgcc aggtaattat accttggact tgtatcatag agatatagat tatacacatt
     2341 tagaaaatag ccaaaagaaa atctagaact agtggatcct aaggtaccta attgcctaga
     2401 aaggagcaga cgatatggcg tcgctccctg caggtcgact ctagaaacca gcagagcata
     2461 tgggctcgct ggctgcagta ttcccgggtt cattagatcc taaggtacct aattgcctag
     2521 aaaggagcag acgatatggc gtcgctccct gcaggtcgac tctagaaacc agcagagcat
     2581 atgggctcgc tggctgcagt attcccgggt tcattagatc ctaaggtacc taattgccta
     2641 gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac cagcagagca
     2701 tatgggctcg ctggctgcag tattcccggg ttcattagat cctaaggtac ctaattgcct
     2761 agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa ccagcagagc
     2821 atatgggctc gctggctgca gtattcccgg gttcattaga tcctaaggta cctaattgcc
     2881 tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa accagcagag
     2941 catatgggct cgctggctgc agtattcccg ggttcattag atcctaaggt acctaattgc
     3001 ctagaaagga gcagacgata tggcgtcgct ccctgcaggt cgactctaga aaccagcaga
     3061 gcatatgggc tcgctggctg cagtattccc gggttcatta gatcctaagg tacctaattg
     3121 cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag aaaccagcag
     3181 agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag gtacctaatt
     3241 gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta gaaaccagca
     3301 gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa ggtacctaat
     3361 tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct agaaaccagc
     3421 agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta aggtacctaa
     3481 ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc tagaaaccag
     3541 cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct aaggtaccta
     3601 attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact ctagaaacca
     3661 gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc atgtcccagg
     3721 atagaatttt gttagataga tctatgccag tgttgaaatc agacaatttc gatccattgg
     3781 aagaagctta cgaaggtggg acaattcaaa actataacga tgaacaccat cttcataaat
     3841 cttgggcaaa tgtgattccg gacaaacgag gactttacga ccctgattat gaacatgacg
     3901 cttgtggtgt cggtttcgta gcaaataagc atggtgaaca gtctcacaag attgttactg
     3961 acgctagata tcttttagtg aatatgacac atcgtggtgc cgtctcatct gatgggaacg
     4021 gtgacggtgc cggtattctg ctaggtatcc caagggcgaa ttcgcggccg ctaaattcaa
     4081 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
     4141 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4201 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctatacgt
     4261 acggcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     4321 acagagtgat attattgaca cgccggggcg acggatggtg atccccctgg ccagtgcacg
     4381 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     4441 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     4501 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     4561 aatataaatg tcaggcatga gattatcaaa aaggatcttc acctagatcc ttttcacgta
     4621 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
     4681 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
     4741 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
     4801 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     4861 ctgatggcgc aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat
     4921 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4981 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5041 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5101 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5161 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5221 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5281 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5341 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5401 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5461 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5521 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5581 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5641 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5701 gttcttctga attattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     5761 gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5821 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagatta tcaaaaagga
     5881 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5941 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6001 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6061 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6121 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6181 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6241 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6301 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6361 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6421 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6481 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6541 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6601 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6661 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6721 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6781 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6841 attgaagcat ttatcagggt tattgtctca tgaccaaaat cccttaacgt gagttttcgt
     6901 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6961 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7021 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7081 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7141 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7201 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7261 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7321 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7381 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7441 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7501 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7561 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7621 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7681 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7741 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     7801 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     7861 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     7921 gaaacagcta tgaccatgat tacgccaagc tcagaattaa ccctcactaa agggactagt
     7981 cctgcaggtt taaacgaatt cgcccttata tattaattaa
//
