LOCUS       sequence-189765-        7819 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 7819)
  AUTHORS   Du Z, Crow ET, Kang HS, Li L
  TITLE     Distinct subregions of Swi1 manifest striking differences in prion 
            transmission and SWI/SNF function.
  JOURNAL   Mol Cell Biol. 2010 Oct;30(19):4644-55. doi: 10.1128/MCB.00225-10. 
            Epub 2010 Aug 2.
  PUBMED    20679490
REFERENCE   2  (bases 1 to 7819)
  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"; doi: 
            "10.1128/MCB.00225-10"; journalName: "Mol Cell Biol"; date: 
            "2010-10"; volume: "30"; issue: "19"; pages: "4644-55"
COMMENT     Sequence Label: p416TEF-NQYFP
FEATURES             Location/Qualifiers
     source          1..7819
                     /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      2212..2221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2218..2220,2221..2223,2224..2937)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2263..2284)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2524..2543)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2871..2892
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2950..3197
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2966..2984)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3215..3234)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3216..3234)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3244..3261)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3244..3260)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3253..3275)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3401..3856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3488..3507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3698..3719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3987..4790)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4791..5006)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5030..5049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5149..5171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5209..5227)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5268..5771
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5808..5912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5913..5981,5982..6773)
                     /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(6131..6150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6944..7532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7433..7452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7686..7703
                     /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 ttctctgaaa gagctgcaat gtttgccgct ttgcagcaaa agcagcagca acgttttcag
     1621 gctctacagc agcagcagca gcagcaacaa aatcaacaac agcaaaatca acaaccacaa
     1681 cagcaacaac agcaacaaca gaatcccaaa tttttgcaga gtcaacggca gcaacaacaa
     1741 agatctattt tacaaagtct gaacccggca ttacaagaga aaatatctac tgaattgaat
     1801 aacaaacaat atgaactttt tatgaaatct ttgattgaaa actgtaaaaa aagaaatatg
     1861 ccgttacaat caataccaga aatcggcaac agaaaaataa atctttttta tctttatatg
     1921 ttggttcaaa aattcggtgg tgcagatcaa gtaacaagga cccaacagtg gtctatggtg
     1981 gcccaaaggc tgcaaatttc cgattaccaa caattagaat caatctattt tagaatttta
     2041 ttaccttacg aaagacacat gatttcccaa gaaggtataa aggaaactca agcgaaaaga
     2101 atattcttgc aacagttctt acaagaacta ttgaagaaag ttcaacaaca gcagcaggcc
     2161 gctgcattgg ccaatgccaa caataacatt aatacccggg atccaccggt cgccaccatg
     2221 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2281 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2341 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2401 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     2461 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2521 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2581 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2641 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2701 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2761 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2821 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2881 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     2941 ggcctcgagt catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     3001 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     3061 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     3121 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     3181 cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     3241 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3301 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3361 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
     3421 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3481 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3541 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3601 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3661 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3721 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3781 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3841 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3901 accgcatagg gtaataactg atataattaa attgaagctc taatttgtga gtttagtata
     3961 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct
     4021 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat
     4081 agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta
     4141 tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac
     4201 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa
     4261 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag
     4321 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct
     4381 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg
     4441 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca
     4501 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt
     4561 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt
     4621 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca
     4681 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca
     4741 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt
     4801 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
     4861 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
     4921 tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa
     4981 gaataaaaaa aaaatgatga attgaaaagg tggtatggtg cactctcagt acaatctgct
     5041 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     5101 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5161 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5221 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacggat cgcttgcctg
     5281 taacttacac gcgcctcgta tcttttaatg atggaataat ttgggaattt actctgtgtt
     5341 tatttatttt tatgttttgt atttggattt tagaaagtaa ataaagaagg tagaagagtt
     5401 acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt
     5461 acatatatat ttattagaca agaaaagcag attaaataga tatacattcg attaacgata
     5521 agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc tacacagaca agatgaaaca
     5581 attcggcatt aatacctgag agcaggaaga gcaagataaa aggtagtatt tgttggcgat
     5641 ccccctagag tcttttacat cttcggaaaa caaaaactat tttttcttta atttcttttt
     5701 ttactttcta tttttaattt atatatttat attaaaaaat ttaaattata attattttta
     5761 tagcacgtga tgaaaaggac ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5821 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5881 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5941 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6001 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6061 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6121 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6181 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6241 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6301 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6361 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6421 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6481 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6541 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6601 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6661 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6721 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6781 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6841 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6901 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6961 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7021 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7081 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7141 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7201 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7261 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7321 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7381 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7441 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7501 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7561 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7621 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7681 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7741 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     7801 gcagtgagcg caacgcaat
//
