LOCUS       sequence-494929-        8146 bp    DNA     circular SYN 09-DEC-2025
DEFINITION  Insertion of the pSTL1 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 8146)
  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 8146)
  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: pSP264
FEATURES             Location/Qualifiers
     source          1..8146
                     /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(4211..4230)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4212..4230)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4237..4254)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4237..4253)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4246..4268)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4391..4693
                     /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     4572..4591
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5042..5836
                     /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             5042..5836
                     /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(5096..5115)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5706..5725
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6086..6877,6878..6946))
                     /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     6709..6728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7070..7658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7559..7578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7812..7829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7946..7967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7982..7987,7988..8005,8006..8012)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8020..8036
                     /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     8025..8047
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8044..8060
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8044..8060
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8081..8099
                     /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 gatcctaagg tacctaattg
     2521 cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag aaaccagcag
     2581 agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag gtacctaatt
     2641 gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta gaaaccagca
     2701 gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa ggtacctaat
     2761 tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct agaaaccagc
     2821 agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta aggtacctaa
     2881 ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc tagaaaccag
     2941 cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct aaggtaccta
     3001 attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact ctagaaacca
     3061 gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc taaggtacct
     3121 aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac tctagaaacc
     3181 agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc ctaaggtacc
     3241 taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac
     3301 cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat cctaaggtac
     3361 ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa
     3421 ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga tcctaaggta
     3481 cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa
     3541 accagcagag catatgggct cgctggctgc agtattcccg ggttcattag atcctaaggt
     3601 acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt cgactctaga
     3661 aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta gatcctaagg
     3721 tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag
     3781 aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt agatccatgt
     3841 cccaggatag aattttgtta gatagatcta tgccagtgtt gaaatcagac aatttcgatc
     3901 cattggaaga agcttacgaa ggtgggacaa ttcaaaacta taacgatgaa caccatcttc
     3961 ataaatcttg ggcaaatgtg attccggaca aacgaggact ttacgaccct gattatgaac
     4021 atgacgcttg tggtgtcggt ttcgtagcaa ataagcatgg tgaacagtct cacaagattg
     4081 ttactgacgc tagatatctt ttagtgaata tgacacatcg tggtgccgtc tcatctgatg
     4141 ggaacggtga cggtgccggt attctgctag gtatcccaag ggcgaattcg cggccgctaa
     4201 attcaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     4261 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     4321 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4381 atacgtacgg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     4441 ggatgtacag agtgatatta ttgacacgcc ggggcgacgg atggtgatcc ccctggccag
     4501 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     4561 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     4621 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     4681 ctggggaata taaatgtcag gcatgagatt atcaaaaagg atcttcacct agatcctttt
     4741 cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc
     4801 tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac
     4861 atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg
     4921 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
     4981 aaggatctga tggcgcaggg gatcaagctc tgatcaagag acaggatgag gatcgtttcg
     5041 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5101 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5161 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5221 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5281 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5341 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5401 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5461 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5521 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     5581 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5641 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5701 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5761 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5821 tgacgagttc ttctgaatta ttaacgctta caatttcctg atgcggtatt ttctccttac
     5881 gcatctgtgc ggtatttcac accgcatcag gtggcacttt tcggggaaat gtgcgcggaa
     5941 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agattatcaa
     6001 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6061 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6121 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6181 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6241 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6301 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6361 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6421 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6481 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6541 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6601 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6661 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6721 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6781 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6841 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6901 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6961 aatattattg aagcatttat cagggttatt gtctcatgac caaaatccct taacgtgagt
     7021 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7081 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7141 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7201 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7261 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7321 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7381 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7441 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7501 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7561 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7621 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7681 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7741 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7801 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7861 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     7921 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     7981 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     8041 acacaggaaa cagctatgac catgattacg ccaagctcag aattaaccct cactaaaggg
     8101 actagtcctg caggtttaaa cgaattcgcc cttatatatt aattaa
//
