LOCUS       sequence-494939-        8339 bp    DNA     circular SYN 09-DEC-2025
DEFINITION  Insertion of the pCTT1 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 8339)
  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 8339)
  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: pSP266
FEATURES             Location/Qualifiers
     source          1..8339
                     /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     complement(4404..4423)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4405..4423)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4430..4447)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4430..4446)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4439..4461)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4584..4886
                     /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     4765..4784
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5235..6029
                     /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             5235..6029
                     /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(5289..5308)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5899..5918
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6279..7070,7071..7139))
                     /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     6902..6921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7263..7851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7752..7771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8005..8022
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8139..8160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8175..8180,8181..8198,8199..8205)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8213..8229
                     /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     8218..8240
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8237..8253
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8237..8253
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8274..8292
                     /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 agagctcttg ccaagtacat agaatccaca gtgcataggt ttatgagatg cttggaagat
     1741 gtacgatcgc ctgcactata ttagtatatt ttttcaggct ttacaaaacc agaaagaaat
     1801 tactgactgt aatacttaat ttccatgatt ttaatcgtat ggtccgtgag gaaagaggaa
     1861 ttttaggtaa aaaaaacctt tgtctatcaa aacataaaag aaaagaaaaa aattaaattg
     1921 aataagtcag ctttttagca tgaccacagt aataatagta atacgatatc agcatgagct
     1981 gctaaacatt aagaattttt gaattgtgtt cgctggtaag agaaaatagg gtagcattag
     2041 ttgcgagatg gaaaattaaa atgtgcgtca taaggtattt tcgacgttga agattagggg
     2101 ttgcaagtcc gagaactaaa ctgccttcac gacgtcaaac ttattgtatg gtgacaataa
     2161 aacgaattca agaagccaga gatgatgagg aaccgatgac gaggcacatg gggatagaac
     2221 ctccgttatc tccattccgc gttgagcagc tctcctgcgt gctttcacag agaccctaag
     2281 ccaagaccgc ggacggccct ctttttcaag gggatcaccg gtaaggggcc aagcggtgaa
     2341 attgcgtatt gtttcctcct ttcaatgatg agtacgtcgc cgatccagaa gggttattat
     2401 tgtcattgcc agtactattc gtatgctgaa aaagtatagt tcacttcacc ctctggctgc
     2461 aggctagcct agccgataat gcccccgact ccctcaacag gtagaatttg taaggattcg
     2521 acgtagcctg gacacattgt gcatttatcg tatcccctac tgctacacgt ataagaagtc
     2581 gttgctttac ctctattcca gaactcaatc ttgtcgttac ttgcccttat taaaaaaatc
     2641 cttctcttgt ctcatgccaa taagatcaat cagctcagct tcacaaacta gtggatccta
     2701 aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc
     2761 tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct
     2821 aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact
     2881 ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc
     2941 taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac
     3001 tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc
     3061 ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga
     3121 ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat
     3181 cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg
     3241 actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga
     3301 tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc
     3361 gactctagaa accagcagag catatgggct cgctggctgc agtattcccg ggttcattag
     3421 atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt
     3481 cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta
     3541 gatcctaagg tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg
     3601 tcgactctag aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt
     3661 agatcctaag gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag
     3721 gtcgactcta gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat
     3781 tagatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca
     3841 ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca
     3901 ttagatccta aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc
     3961 aggtcgactc tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc
     4021 attagatcca tgtcccagga tagaattttg ttagatagat ctatgccagt gttgaaatca
     4081 gacaatttcg atccattgga agaagcttac gaaggtggga caattcaaaa ctataacgat
     4141 gaacaccatc ttcataaatc ttgggcaaat gtgattccgg acaaacgagg actttacgac
     4201 cctgattatg aacatgacgc ttgtggtgtc ggtttcgtag caaataagca tggtgaacag
     4261 tctcacaaga ttgttactga cgctagatat cttttagtga atatgacaca tcgtggtgcc
     4321 gtctcatctg atgggaacgg tgacggtgcc ggtattctgc taggtatccc aagggcgaat
     4381 tcgcggccgc taaattcaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     4441 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4501 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4561 agttgcgcag cctatacgta cggcagttta aggtttacac ctataaaaga gagagccgtt
     4621 atcgtctgtt tgtggatgta cagagtgata ttattgacac gccggggcga cggatggtga
     4681 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     4741 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     4801 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     4861 ttaacctgat gttctgggga atataaatgt caggcatgag attatcaaaa aggatcttca
     4921 cctagatcct tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
     4981 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
     5041 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
     5101 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
     5161 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag ctctgatcaa gagacaggat
     5221 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5281 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5341 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5401 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5461 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5521 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5581 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5641 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5701 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5761 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5821 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5881 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5941 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6001 tctatcgcct tcttgacgag ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt
     6061 attttctcct tacgcatctg tgcggtattt cacaccgcat caggtggcac ttttcgggga
     6121 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6181 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6241 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6301 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6361 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6421 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6481 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6541 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6601 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6661 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6721 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6781 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6841 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6901 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6961 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7021 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7081 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7141 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gaccaaaatc
     7201 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7261 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7321 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7381 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7441 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7501 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7561 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7621 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7681 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7741 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7801 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7861 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7921 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7981 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     8041 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     8101 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     8161 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     8221 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cagaattaac
     8281 cctcactaaa gggactagtc ctgcaggttt aaacgaattc gcccttatat attaattaa
//
