LOCUS       sequence-269890-        8461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Repair DNA plasmid to insert PP7 stem loops in the endogenous STL1 
            locus after double strand break in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8461)
  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 8461)
  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: pSP569
FEATURES             Location/Qualifiers
     source          1..8461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        416..759
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             760..1458
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1462..1659
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     2496..2512
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2496..2512
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4536..4555)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4537..4555)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4562..4579)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4562..4578)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4571..4593)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4716..5018
                     /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     4897..4916
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5367..6161
                     /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             5367..6161
                     /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(5421..5440)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6031..6050
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(6707..7085)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     7032..7051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7393..7981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7882..7901
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8135..8152
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8269..8290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8305..8310,8311..8328,8329..8335)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8343..8359
                     /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     8348..8370
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8367..8383
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8367..8383
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8404..8422
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaattaaac cgttcaaccc tatacagcgc aaatttggtc ctaatacact tcggttttaa
       61 tgcgtcaatc cgattggctc cgataagctt ttgcacattt ttcaagtcat atgtcacgac
      121 gaacgtgatg cccgcaaaac gtaaaaaaaa gaagcaataa attgcccaaa aaaaaaatat
      181 aaactgcttt tcttagtcat tcataagttc tgatggtctt ccttatctta ttgtattttt
      241 cgtcctattg tttcatttct taccaaatta atcaattctt atatcttact tgataacaca
      301 ccaaactaat cgtctccaca tcataggaag ataggaaatt gctatctcag tcctatacta
      361 cgcagacctc gagtctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat
      421 ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga
      481 ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt
      541 tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg
      601 aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat
      661 aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct
      721 aggatacagt tctcacatca catccgaaca taaacaacca tggcagaacc agcccaaaaa
      781 aagcaaaaac aaactgttca ggagcgcaag gcgtttatct cccgtatcac taatgaaact
      841 aaaattcaaa tcgctatttc gctgaatggt ggttatattc aaataaaaga ttcgattctt
      901 cctgcaaaga aggatgacga tgtagcttcc caagctactc agtcacaggt catcgatatt
      961 cacacaggtg ttggcttttt ggatcatatg atccatgcgt tggcaaaaca ctctggttgg
     1021 tctcttattg ttgaatgtat tggtgacctg cacattgacg atcaccatac taccgaagat
     1081 tgcggtatcg cattagggca agcgttcaaa gaagcaatgg gtgctgtccg tggtgtaaaa
     1141 agattcggta ctgggttcgc accattggat gaggcgctat cacgtgccgt agtcgattta
     1201 tctagtagac catttgctgt aatcgacctt ggattgaaga gagagatgat tggtgattta
     1261 tccactgaaa tgattccaca ctttttggaa agtttcgcgg aggcggccag aattactttg
     1321 catgttgatt gtctgagagg tttcaacgat caccacagaa gtgagagtgc gttcaaggct
     1381 ttggctgttg ccataagaga agctatttct agcaatggca ccaatgacgt tccctcaacc
     1441 aaaggtgttt tgatgtgaag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     1501 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     1561 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     1621 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     1681 cgccatccag agctcgcaga acaccactac tgatactcat tgccaaggct aggaggcacc
     1741 atccgtttca ttttctttga ggtaagccaa tcatgaaata gtatacacat ccataacgga
     1801 cgtacggacg aaataagtgc cgttgtccca ctattccacc gcatttggcc catttggctc
     1861 actttgactc aacttgcgtc attttaactg atatgaaggg tccgactttg tcctttttcg
     1921 gccaccgcat accccacggc gatgcctccg ctacctgcat ttgagtaaca tctccgtttc
     1981 gcggggtatt cggcgctacg tcgcctgttc gagcggctct gttcgttgca tgaaactaaa
     2041 ataagcggaa agtgtccagc catccactac gtcagaaaga aataatggtt gtacactgtt
     2101 tctcggctat ataccgtttt tggttggtta atcctcgcca ggtgcagcta ttgcgcttgg
     2161 ctgcttcgcg atagtagtaa tctgagaaag tgcagatccc ggtaagggaa acacttttgg
     2221 ttcacctttg atagggcttt cattggggca ttcgtaacaa aaaggaagta gatagagaaa
     2281 ttgagaaagc ttaagtgaga tgttttagct tcaattttgt ccccttcaac gctgcttggc
     2341 cttagagggt cagaattgca gttcaggagt agtcacactc atagtatata aacaagccct
     2401 ttattgattt tgaataatta ttttgtatac gtgttctagc atacaagtta gaataaataa
     2461 aaaatagaaa aatagaacat agaaagtttt agacctctag aactagtgga tcctaaggta
     2521 cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa
     2581 accagcagag catatgggct cgctggctgc agtattcccg ggttcattag atcctaaggt
     2641 acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt cgactctaga
     2701 aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta gatcctaagg
     2761 tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag
     2821 aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag
     2881 gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta
     2941 gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa
     3001 ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct
     3061 agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta
     3121 aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc
     3181 tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct
     3241 aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact
     3301 ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc
     3361 taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac
     3421 tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc
     3481 ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga
     3541 ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat
     3601 cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg
     3661 actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga
     3721 tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc
     3781 gactctagaa accagcagag catatgggct cgctggctgc agtattcccg ggttcattag
     3841 atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt
     3901 cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta
     3961 gatccatgtc ccaggataga attttgttag atagatctga tatccgcatc tatgacgggc
     4021 ttctccctgt ttggatacga ccaagggttg atggcaagtc taattactgg taaacagttc
     4081 aactatgaat ttccagcaac caaagaaaat ggcgatcatg acagacacgc aactgtagtg
     4141 cagggcgcta caacctcctg ttatgaatta ggttgtttcg caggttctct attcgttatg
     4201 ttctgcggtg aaagaattgg tagaaaacca ttaatcctga tgggttccgt aataaccatc
     4261 attggtgccg ttatttctac atgcgcattt cgtggttact gggcattagg ccagtttatc
     4321 atcggaagag tcgtcaccgg tgttggaaca gggttgaata catctactat tcccgtttgg
     4381 caatcagaaa tgtcaaaagc tgaaaataga gggttgctgg tcaatttaga aggttccaca
     4441 attgcttttg gtactatgat tgcttattgg attgattttg ggttgtctta taccaacagt
     4501 tctgttcagt ggaggcggcc gctaaattca attcgcccta tagtgagtcg tattacaatt
     4561 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4621 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4681 gcccttccca acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa
     4741 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc
     4801 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac
     4861 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca
     4921 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca
     4981 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggcatg agattatcaa
     5041 aaaggatctt cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac
     5101 cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa
     5161 agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag
     5221 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag
     5281 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agctctgatc
     5341 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5401 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5461 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5521 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5581 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5641 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5701 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5761 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5821 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5881 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5941 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6001 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6061 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6121 agcgcatcgc cttctatcgc cttcttgacg agttcttctg aattattaac gcttacaatt
     6181 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atcaggtggc
     6241 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6301 atgtatccgc tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6541 atgataccgc gagaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6601 gaaggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6661 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6721 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6781 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6841 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6901 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6961 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7021 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7081 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7141 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7201 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7261 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7321 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7381 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7441 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7501 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7561 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7621 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7681 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7741 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7801 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7861 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7921 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7981 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8041 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     8101 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     8161 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     8221 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     8281 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     8341 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     8401 cagaattaac cctcactaaa gggactagtc ctgcaggttt aaacgaattc gcccttatat
     8461 a
//
