LOCUS       sequence-189755-        8316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Swi1 NQ-YFP to monitor Swi1 state or induce [SWI+]..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8316)
  AUTHORS   Hines JK, Li X, Du Z, Higurashi T, Li L, Craig EA
  TITLE     [SWI], the prion formed by the chromatin remodeling factor Swi1, is 
            highly sensitive to alterations in Hsp70 chaperone system activity.
  JOURNAL   PLoS Genet. 2011 Feb;7(2):e1001309. doi: 
            10.1371/journal.pgen.1001309. Epub 2011 Feb 17.
  PUBMED    21379326
REFERENCE   2  (bases 1 to 8316)
  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: "PLoS 
            Genet."; date: "2011-02"; volume: "7(2):e1001309. doi"; pages: " 
            10.1371/journal.pgen.1001309. Epub 2011 Feb 17"
COMMENT     Sequence Label: p415TEF-NYFP
FEATURES             Location/Qualifiers
     source          1..8316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     134..154
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        172..570
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     regulatory      1585..1594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1591..1593,1594..1596,1597..2310)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1636..1657)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1897..1916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2244..2265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2323..2570
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2339..2357)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2588..2607)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2589..2607)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2617..2634)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2617..2633)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2626..2648)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2774..3229
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2861..2880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3071..3092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3530..3937
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3938..5032
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(5526..5545)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5645..5667
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5705..5723)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5765..6268
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6305..6409
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6410..6478,6479..7270)
                     /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     complement(6628..6647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7441..8029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7930..7949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8183..8200
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tcatagcttc
      181 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
      241 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
      301 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc
      361 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
      421 tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat
      481 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat
      541 tagaaagaaa gcatagcaat ctaatctaag ttttctagaa ctagtatgga tttctttaat
      601 ttgaataata ataataataa taataatact actactacta ctactactac caataacaat
      661 aatactaata ataataatac taataataat aataatcctg ctaataatac caataacaac
      721 aatagcacag gccatagtag taatactaat aacaatacta ataacaacaa tacgaatacc
      781 ggagccagtg gtgtagacga ttttcagaat tttttcgacc caaaaccttt tgaccagaat
      841 ttggattcca acaataacaa cagtaatagt aataacaatg ataacaataa cagtaacacg
      901 gtagcctcca gcacgaactt tacttctcca acagcagtcg ttaataatgc cgctcctgct
      961 aatgttaccg gtggaaaagc tgctaatttc attcagaacc agtctcccca attcaattct
     1021 ccatatgact caaacaattc caacacaaat ttaaacagcc tgtctcctca ggctatttta
     1081 gccaagaatt caatcattga ttcttccaat ttacctcttc aagctcaaca acagctatat
     1141 ggcggtaaca ataataataa tagcaccgga attgcgaacg ataatgtaat aacaccccat
     1201 tttatcacca atgttcaatc catcagtcaa aattcctcat cttctactcc gaacacaaac
     1261 tctaactcta ctccgaatgc aaatcaacaa tttttgccat tcaataacag tgcttccaat
     1321 aatggtaatt tgacgtctaa ccagcttatt tctaattacg ctgcttcaaa ttcaatggac
     1381 agatcatcct ctgcaagcaa tgaatttgtt ccgaacacaa gtgacaacaa caacaacagt
     1441 aataaccata atatgcgtaa taacagcaat aataaaacga gcaataacaa taatgtcact
     1501 gccgtgcctg ctgcaactcc cgctaatacc aataattcaa cttcaaacgc aaatacagtg
     1561 ttctctgccc gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     1621 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1681 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1741 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag
     1801 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1861 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1921 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1981 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2041 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2101 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2161 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     2221 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2281 actctcggca tggacgagct gtacaagtaa agcggcctcg agtcatgtaa ttagttatgt
     2341 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     2401 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     2461 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     2521 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtacc
     2581 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     2641 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     2701 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     2761 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2821 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2881 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2941 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3001 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3061 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3121 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3181 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     3241 attttctcct tacgcatctg tgcggtattt cacaccgcat atcgaccggt cgaggagaac
     3301 ttctagtata tctacatacc taatattatt gccttattaa aaatggaatc ccaacaatta
     3361 catcaaaatc cacattctct tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt
     3421 caaaaacgtt atatttatag gataattata ctctatttct caacaagtaa ttggttgttt
     3481 ggccgagcgg tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa
     3541 tactcaggta tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc
     3601 aacataacga gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt
     3661 ttttgttaat ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga
     3721 aaaaggaaag gtgagagcgc cggaaccggc ttttcatata gaatagagaa gcgttcatga
     3781 ctaaatgctt gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt
     3841 tccaataggt ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc
     3901 aatatatata tatatatttc aaggatatac cattctaatg tctgccccta agaagatcgt
     3961 cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc gaagccatta aggttcttaa
     4021 agctatttct gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg
     4081 tgctgctatc gatgctacag gtgttccact tccagatgag gcgctggaag cctccaagaa
     4141 ggctgatgcc gttttgttag gtgctgtggg tggtcctaaa tggggtaccg gtagtgttag
     4201 acctgaacaa ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca acttaagacc
     4261 atgtaacttt gcatccgact ctcttttaga cttatctcca atcaagccac aatttgctaa
     4321 aggtactgac ttcgttgttg tcagagaatt agtgggaggt atttactttg gtaagagaaa
     4381 ggaagacgat ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc cagaagtgca
     4441 aagaatcaca agaatggccg ctttcatggc cctacaacat gagccaccat tgcctatttg
     4501 gtccttggat aaagctaatg ttttggcctc ttcaagatta tggagaaaaa ctgtggagga
     4561 aaccatcaag aacgaattcc ctacattgaa ggttcaacat caattgattg attctgccgc
     4621 catgatccta gttaagaacc caacccacct aaatggtatt ataatcacca gcaacatgtt
     4681 tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt tgttgccatc
     4741 tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg aaccatgcca
     4801 cggttctgct ccagatttgc caaagaataa ggtcaaccct atcgccacta tcttgtctgc
     4861 tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc
     4921 agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt ccaacagtac
     4981 caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccttgctt aaaaagattc
     5041 tcttttttta tgatatttgt acataaactt tataaatgaa attcataata gaaacgacac
     5101 gaaattacaa aatggaatat gttcataggg tagacgaaac tatatacgca atctacatac
     5161 atttatcaag aaggagaaaa aggaggatgt aaaggaatac aggtaagcaa attgatacta
     5221 atggctcaac gtgataagga aaaagaattg cactttaaca ttaatattga caaggaggag
     5281 ggcaccacac aaaaagttag gtgtaacaga aaatcatgaa actatgattc ctaatttata
     5341 tattggagga ttttctctaa aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct
     5401 gaccatttga tggagtttaa gtcaatacct tcttgaacca tttcccataa tggtgaaagt
     5461 tccctcaaga attttactct gtcagaaacg gccttaacga cgtagtcgat atggtgcact
     5521 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5581 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5641 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5701 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5761 gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg
     5821 ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata
     5881 aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc
     5941 aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat
     6001 acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac
     6061 acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg
     6121 tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt
     6181 ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta
     6241 aattataatt atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa
     6301 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6361 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6421 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6481 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6541 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6601 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6661 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6721 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6781 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6841 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6901 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6961 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7021 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7081 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7141 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7201 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7261 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7321 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7381 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7441 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7501 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7561 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7621 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7681 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7741 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7801 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7861 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7921 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7981 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8041 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8101 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8161 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     8221 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     8281 tcccgactgg aaagcgggca gtgagcgcaa cgcaat
//
