LOCUS       sequence-494931-        8346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insertion of the pGPD1 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: pVW200
FEATURES             Location/Qualifiers
     source          1..8346
                     /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     2688..2704
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2688..2704
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4411..4430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4412..4430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4437..4454)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4437..4453)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4446..4468)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4591..4893
                     /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     4772..4791
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5242..6036
                     /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             5242..6036
                     /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(5296..5315)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5906..5925
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6286..7077,7078..7146))
                     /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     6909..6928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7270..7858
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8012..8029
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8146..8167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8182..8187,8188..8205,8206..8212)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8220..8236
                     /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     8225..8247
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8244..8260
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8244..8260
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8281..8299
                     /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 agagctcgaa gcccgaaaga gttatcgtta ctccgattat tttgtacagc tgatgggacc
     1741 ttgccgtctt catttttttt ttttcaccta tagagccggg cagagctgcc cggcttaact
     1801 aagggccgga aaaaaaacgg aaaaaagaaa gccaagcgtg tagacgtagt ataacagtat
     1861 atctgacacg cacgtgatga ccacgtaatc gcatcgcccc tcacctctca tctctcaccg
     1921 ctgactcagc ttcactaaaa aggaaaatat atactctttc ccaggcaagg tgacagcggt
     1981 ccccgtctcc tccacaaagg cctctcctgg ggtttgagca agtctaagtt tacgtagcat
     2041 aaaaattctc gcattgcgtc aaataataaa aaaagtaacc ccacttctac ttctacatcg
     2101 gaaaaacatt ccattcacat atcgtctttg gcctatcttg ttttgtcctc ggtagatcag
     2161 gtcagtacaa acgcaacacg aaagaacaaa aaaagaagaa aaacagaagg ccaagacagg
     2221 gtcaatgaga ctgttgtcct cctactgtcc ctatgtctct ggccgatcac gcgccattgt
     2281 ccctcagaaa caaatcaaac acccacaccc ctggcaccca aagtccccac ccacaccacc
     2341 aatacgtaaa cggggcgccc cctgcaaggc cctcctgcgc gcggcctccc gccttgcttc
     2401 tctcccctcc cttttctttt tccagttttc cctattttgt ccctttttcc gcacaacaag
     2461 tatcagaatg ggttcatcaa atctatccaa cctaattcgc acgtagactg gcttggtatt
     2521 ggcagtttcg cagttatata tatactacca tgagtgaaac tgttacgtta ccttaaattc
     2581 tttctccctt taattttctt ttatcttact ctcctacata agacatcaag aaacaattgt
     2641 atattgtaca ccccccctcc acaaacacaa atattgataa tataaagtct agaactagtg
     2701 gatcctaagg tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg
     2761 tcgactctag aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt
     2821 agatcctaag gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag
     2881 gtcgactcta gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat
     2941 tagatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca
     3001 ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca
     3061 ttagatccta aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc
     3121 aggtcgactc tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc
     3181 attagatcct aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg
     3241 caggtcgact ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt
     3301 cattagatcc taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct
     3361 gcaggtcgac tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt
     3421 tcattagatc ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc
     3481 tgcaggtcga ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg
     3541 ttcattagat cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc
     3601 ctgcaggtcg actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg
     3661 gttcattaga tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc
     3721 cctgcaggtc gactctagaa accagcagag catatgggct cgctggctgc agtattcccg
     3781 ggttcattag atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct
     3841 ccctgcaggt cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc
     3901 gggttcatta gatcctaagg tacctaattg cctagaaagg agcagacgat atggcgtcgc
     3961 tccctgcagg tcgactctag aaaccagcag agcatatggg ctcgctggct gcagtattcc
     4021 cgggttcatt agatccatgt cccaggatag aattttgtta gatagatcta tgccagtgtt
     4081 gaaatcagac aatttcgatc cattggaaga agcttacgaa ggtgggacaa ttcaaaacta
     4141 taacgatgaa caccatcttc ataaatcttg ggcaaatgtg attccggaca aacgaggact
     4201 ttacgaccct gattatgaac atgacgcttg tggtgtcggt ttcgtagcaa ataagcatgg
     4261 tgaacagtct cacaagattg ttactgacgc tagatatctt ttagtgaata tgacacatcg
     4321 tggtgccgtc tcatctgatg ggaacggtga cggtgccggt attctgctag gtatcccaag
     4381 ggcgaattcg cggccgctaa attcaattcg ccctatagtg agtcgtatta caattcactg
     4441 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4501 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4561 tcccaacagt tgcgcagcct atacgtacgg cagtttaagg tttacaccta taaaagagag
     4621 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc ggggcgacgg
     4681 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac
     4741 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     4801 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     4861 aacgccatta acctgatgtt ctggggaata taaatgtcag gcatgagatt atcaaaaagg
     4921 atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg
     4981 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag
     5041 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc
     5101 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     5161 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagctc tgatcaagag
     5221 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5281 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5341 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5401 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     5461 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5521 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5581 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5641 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5701 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5761 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5821 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5881 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5941 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6001 atcgccttct atcgccttct tgacgagttc ttctgaatta ttaacgctta caatttcctg
     6061 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt
     6121 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6181 tccgctcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgac
     7201 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7261 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7321 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7381 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7441 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7501 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7561 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7621 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7681 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7741 cacgagggag cttccagggg gaaacgcctg ctatctttat agtcctgtcg ggtttcgcca
     7801 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7861 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7921 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7981 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     8041 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     8101 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     8161 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     8221 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcag
     8281 aattaaccct cactaaaggg actagtcctg caggtttaaa cgaattcgcc cttatatatt
     8341 aattaa
//
