LOCUS       sequence-493898-        8341 bp    DNA     circular SYN 15-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 8341)
  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 8341)
  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: pVW293
FEATURES             Location/Qualifiers
     source          1..8341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        391..734
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             735..1433
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1437..1634
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4389..4408)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4390..4408)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4415..4432)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4415..4431)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4424..4446)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4569..4871
                     /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     4750..4769
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5220..6014
                     /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             5220..6014
                     /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(5274..5293)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5884..5903
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6264..7055,7056..7124))
                     /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     6887..6906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7248..7836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7737..7756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7990..8007
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8124..8145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8160..8165,8166..8183,8184..8190)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8198..8214
                     /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     8203..8225
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8222..8238
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8222..8238
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8259..8277
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcgcaaatt tggtcctaat acacttcggt tttaatgcgt caatccgatt ggctccgata
       61 agcttttgca catttttcaa gtcatatgtc acgacgaacg tgatgcccgc aaaacgtaaa
      121 aaaaagaagc aataaattgc ccaaaaaaaa aatataaact gcttttctta gtcattcata
      181 agttctgatg gtcttcctta tcttattgta tttttcgtcc tattgtttca tttcttacca
      241 aattaatcaa ttcttatatc ttacttgata acacaccaaa ctaatcgtct ccacatcata
      301 ggaagatagg aaattgctat ctcagtccta tactacgcag acctcgagtc tgtttagctt
      361 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
      421 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
      481 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
      541 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
      601 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
      661 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
      721 gaacataaac aaccatggca gaaccagccc aaaaaaagca aaaacaaact gttcaggagc
      781 gcaaggcgtt tatctcccgt atcactaatg aaactaaaat tcaaatcgct atttcgctga
      841 atggtggtta tattcaaata aaagattcga ttcttcctgc aaagaaggat gacgatgtag
      901 cttcccaagc tactcagtca caggtcatcg atattcacac aggtgttggc tttttggatc
      961 atatgatcca tgcgttggca aaacactctg gttggtctct tattgttgaa tgtattggtg
     1021 acctgcacat tgacgatcac catactaccg aagattgcgg tatcgcatta gggcaagcgt
     1081 tcaaagaagc aatgggtgct gtccgtggtg taaaaagatt cggtactggg ttcgcaccat
     1141 tggatgaggc gctatcacgt gccgtagtcg atttatctag tagaccattt gctgtaatcg
     1201 accttggatt gaagagagag atgattggtg atttatccac tgaaatgatt ccacactttt
     1261 tggaaagttt cgcggaggcg gccagaatta ctttgcatgt tgattgtctg agaggtttca
     1321 acgatcacca cagaagtgag agtgcgttca aggctttggc tgttgccata agagaagcta
     1381 tttctagcaa tggcaccaat gacgttccct caaccaaagg tgttttgatg tgaagtactg
     1441 acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag
     1501 ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct
     1561 gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
     1621 tggtcgctat actgctgtcg attcgatact aacgccgcca tccagagctc tgatatagga
     1681 cttctctctc tttttttttt ttttttgtcc aggtggagtg cgatttgttc gttatatgcc
     1741 tcattcctta atttttatca gtcagaacgt tatttcgcgc tgcaagttcc ttacttttcc
     1801 atagagttcc acaggatgta aagaaaaata atgataaaaa aaagcagaac atattaccct
     1861 tcttttggag cgtacttcca tctgttaccc gctcatccaa atgcacattc tgttaccctc
     1921 ttgttcggta tttttcggag ttctcctttt ctccttcttt tcctcttgat acacagaaaa
     1981 gtgggagaag gtagggggag tcagagaatg aagaaaaaaa ggggacgcgc aaatccaagt
     2041 gaaaatctcc gggagcgggc ggatcccact aacggcccag ccgaaaatgg aaaaaaaggg
     2101 tcggtgatgt gtgggtgcca gctggcggta gcaatgacga cgtgttgacg ggcccttggc
     2161 tcttgggaca aggactagaa gccaaaagcc agaggcggta aaaatagcaa gactagaata
     2221 ttgctggcat ctgttaaggg gatatgttgc aacttgcagg gggcggcaca aaataacata
     2281 gaaacgtagt aaagagggga aaaggaaaag gaaaaggaaa aggaaggaaa aaaacccatt
     2341 gacgtagaaa ttgaaagaag gaaaggtata cgcaagcatt aatacaaccc acaaacacag
     2401 accagaagca ctctagacgg agagtaacta gatctacagc ccctggaaaa tcgtttggtc
     2461 aactttgagg ttccggtcgt ccccctcttg atctgaaagg tctttctcta aatctatatt
     2521 aaaacgtata aataggacgg tgaattgcgt tctacttcct caattgcgtt tgatcttatt
     2581 taatctctct ctaatatata gaaaaaaaaa ccatctgatt attcgataat ctcaaacaaa
     2641 caactcaaaa caaaaaaaac taaatacaac accgcgggga tcctaaggta cctaattgcc
     2701 tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa accagcagag
     2761 catatgggct cgctggctgc agtattcccg ggttcattag atcctaaggt acctaattgc
     2821 ctagaaagga gcagacgata tggcgtcgct ccctgcaggt cgactctaga aaccagcaga
     2881 gcatatgggc tcgctggctg cagtattccc gggttcatta gatcctaagg tacctaattg
     2941 cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag aaaccagcag
     3001 agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag gtacctaatt
     3061 gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta gaaaccagca
     3121 gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa ggtacctaat
     3181 tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct agaaaccagc
     3241 agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta aggtacctaa
     3301 ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc tagaaaccag
     3361 cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct aaggtaccta
     3421 attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact ctagaaacca
     3481 gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc taaggtacct
     3541 aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac tctagaaacc
     3601 agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc ctaaggtacc
     3661 taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac
     3721 cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat cctaaggtac
     3781 ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa
     3841 ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga tcctaaggta
     3901 cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa
     3961 accagcagag catatgggct cgctggctgc agtattcccg ggttcattag atccatgtcc
     4021 caggatagaa ttttgttaga tagatctatg ccagtgttga aatcagacaa tttcgatcca
     4081 ttggaagaag cttacgaagg tgggacaatt caaaactata acgatgaaca ccatcttcat
     4141 aaatcttggg caaatgtgat tccggacaaa cgaggacttt acgaccctga ttatgaacat
     4201 gacgcttgtg gtgtcggttt cgtagcaaat aagcatggtg aacagtctca caagattgtt
     4261 actgacgcta gatatctttt agtgaatatg acacatcgtg gtgccgtctc atctgatggg
     4321 aacggtgacg gtgccggtat tctgctaggt atcccaaggg cgaattcgcg gccgctaaat
     4381 tcaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
     4441 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     4501 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctat
     4561 acgtacggca gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg
     4621 atgtacagag tgatattatt gacacgccgg ggcgacggat ggtgatcccc ctggccagtg
     4681 cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg
     4741 aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag
     4801 aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct
     4861 ggggaatata aatgtcaggc atgagattat caaaaaggat cttcacctag atccttttca
     4921 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta
     4981 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat
     5041 ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg
     5101 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa
     5161 ggatctgatg gcgcagggga tcaagctctg atcaagagac aggatgagga tcgtttcgca
     5221 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5281 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5341 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5401 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5461 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5521 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5581 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5641 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5701 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     5761 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5821 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5881 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5941 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6001 acgagttctt ctgaattatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     6061 atctgtgcgg tatttcacac cgcatcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6121 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag attatcaaaa
     6181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7141 tattattgaa gcatttatca gggttattgt ctcatgacca aaatccctta acgtgagttt
     7201 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7261 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7321 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7381 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7441 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7501 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7561 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7621 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7681 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7741 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7801 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7861 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7921 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7981 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     8041 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     8101 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     8161 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     8221 acaggaaaca gctatgacca tgattacgcc aagctcagaa ttaaccctca ctaaagggac
     8281 tagtcctgca ggtttaaacg aattcgccct tatatattaa ttaaaccgtt caaccctata
     8341 c
//
