LOCUS       sequence-494933-        8078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insertion of the pSTL1 promoter and 24 MS2 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 8078)
  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 8078)
  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: pSP568
FEATURES             Location/Qualifiers
     source          1..8078
                     /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     2488..2504
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2488..2504
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4143..4162)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4144..4162)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4169..4186)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4169..4185)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4178..4200)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4323..4625
                     /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     4504..4523
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             4974..5768
                     /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             4974..5768
                     /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(5028..5047)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5638..5657
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6018..6809,6810..6878))
                     /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     6641..6660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7002..7590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7491..7510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7744..7761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7878..7899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7914..7919,7920..7937,7938..7944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7952..7968
                     /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     7957..7979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7976..7992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7976..7992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8013..8031
                     /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 agagctcgca gaacaccact actgatactc attgccaagg ctaggaggca ccatccgttt
     1741 cattttcttt gaggtaagcc aatcatgaaa tagtatacac atccataacg gacgtacgga
     1801 cgaaataagt gccgttgtcc cactattcca ccgcatttgg cccatttggc tcactttgac
     1861 tcaacttgcg tcattttaac tgatatgaag ggtccgactt tgtccttttt cggccaccgc
     1921 ataccccacg gcgatgcctc cgctacctgc atttgagtaa catctccgtt tcgcggggta
     1981 ttcggcgcta cgtcgcctgt tcgagcggct ctgttcgttg catgaaacta aaataagcgg
     2041 aaagtgtcca gccatccact acgtcagaaa gaaataatgg ttgtacactg tttctcggct
     2101 atataccgtt tttggttggt taatcctcgc caggtgcagc tattgcgctt ggctgcttcg
     2161 cgatagtagt aatctgagaa agtgcagatc ccggtaaggg aaacactttt ggttcacctt
     2221 tgatagggct ttcattgggg cattcgtaac aaaaaggaag tagatagaga aattgagaaa
     2281 gcttaagtga gatgttttag cttcaatttt gtccccttca acgctgcttg gccttagagg
     2341 gtcagaattg cagttcagga gtagtcacac tcatagtata taaacaagcc ctttattgat
     2401 tttgaataat tattttgtat acgtgttcta gcatacaagt tagaataaat aaaaaataga
     2461 aaaatagaac atagaaagtt ttagacctct agaactagtg gatcctacgg tacttattgc
     2521 caagaaagca cgagcatcag ccgtgcctcc aggtcgaatc ttcaaacgac gacgatcacg
     2581 cgtcgctcca gtattccagg gttcatcaga tcctacggta cttattgcca agaaagcacg
     2641 agcatcagcc gtgcctccag gtcgaatctt caaacgacga cgatcacgcg tcgctccagt
     2701 attccagggt tcatcagatc ctacggtact tattgccaag aaagcacgag catcagccgt
     2761 gcctccaggt cgaatcttca aacgacgacg atcacgcgtc gctccagtat tccagggttc
     2821 atcagatcct acggtactta ttgccaagaa agcacgagca tcagccgtgc ctccaggtcg
     2881 aatcttcaaa cgacgacgat cacgcgtcgc tccagtattc cagggttcat cagatcctac
     2941 ggtacttatt gccaagaaag cacgagcatc agccgtgcct ccaggtcgaa tcttcaaacg
     3001 acgacgatca cgcgtcgctc cagtattcca gggttcatca gatcctacgg tacttattgc
     3061 caagaaagca cgagcatcag ccgtgcctcc aggtcgaatc ttcaaacgac gacgatcacg
     3121 cgtcgctcca gtattccagg gttcatcaga tcctacggta cttattgcca agaaagcacg
     3181 agcatcagcc gtgcctccag gtcgaatctt caaacgacga cgatcacgcg tcgctccagt
     3241 attccagggt tcatcagatc ctacggtact tattgccaag aaagcacgag catcagccgt
     3301 gcctccaggt cgaatcttca aacgacgacg atcacgcgtc gctccagtat tccagggttc
     3361 atcagatcct acggtactta ttgccaagaa agcacgagca tcagccgtgc ctccaggtcg
     3421 aatcttcaaa cgacgacgat cacgcgtcgc tccagtattc cagggttcat cagatcctac
     3481 ggtacttatt gccaagaaag cacgagcatc agccgtgcct ccaggtcgaa tcttcaaacg
     3541 acgacgatca cgcgtcgctc cagtattcca gggttcatca gatcctacgg tacttattgc
     3601 caagaaagca cgagcatcag ccgtgcctcc aggtcgaatc ttcaaacgac gacgatcacg
     3661 cgtcgctcca gtattccagg gttcatcaga tcctacggta cttattgcca agaaagcacg
     3721 agcatcagcc gtgcctccag gtcgaatctt caaacgacga cgatcacgcg tcgctccagt
     3781 attccagggt tcatcagatc tatgccagtg ttgaaatcag acaatttcga tccattggaa
     3841 gaagcttacg aaggtgggac aattcaaaac tataacgatg aacaccatct tcataaatct
     3901 tgggcaaatg tgattccgga caaacgagga ctttacgacc ctgattatga acatgacgct
     3961 tgtggtgtcg gtttcgtagc aaataagcat ggtgaacagt ctcacaagat tgttactgac
     4021 gctagatatc ttttagtgaa tatgacacat cgtggtgccg tctcatctga tgggaacggt
     4081 gacggtgccg gtattctgct aggtatccca agggcgaatt cgcggccgct aaattcaatt
     4141 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     4201 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4261 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctatacgtac
     4321 ggcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     4381 agagtgatat tattgacacg ccggggcgac ggatggtgat ccccctggcc agtgcacgtc
     4441 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     4501 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     4561 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     4621 tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga
     4681 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     4741 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     4801 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     4861 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     4921 gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4981 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5041 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5101 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5161 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5221 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5281 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5341 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5401 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5461 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5521 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5581 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5641 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5701 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5761 tcttctgaat tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     5821 gcggtatttc acaccgcatc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5881 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagattatc aaaaaggatc
     5941 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6001 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6061 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6241 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6301 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6421 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6541 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6661 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6841 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6901 tgaagcattt atcagggtta ttgtctcatg accaaaatcc cttaacgtga gttttcgttc
     6961 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7021 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7081 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7141 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7201 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7261 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7321 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7381 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7441 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7501 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7561 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7621 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7681 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7741 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7801 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     7861 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     7921 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     7981 aacagctatg accatgatta cgccaagctc agaattaacc ctcactaaag ggactagtcc
     8041 tgcaggttta aacgaattcg cccttatata ttaattaa
//
