LOCUS       sequence-189770-        7192 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 7192)
  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 7192)
  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-NYFP
FEATURES             Location/Qualifiers
     source          1..7192
                     /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"
     CDS             complement(3360..4163)
                     /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(4164..4379)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4403..4422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4522..4544
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4582..4600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4641..5144
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5181..5285
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5286..5354,5355..6146)
                     /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(5504..5523)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6317..6905
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6806..6825
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7059..7076
                     /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 agggtaataa ctgatataat
     3301 taaattgaag ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt
     3361 tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca
     3421 ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca
     3481 gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg
     3541 tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc
     3601 atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca
     3661 gtacccttag tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc
     3721 aaaaggcctc taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata
     3781 cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca
     3841 cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag
     3901 agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa
     3961 aaatcagtca agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca
     4021 actaactcca gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc
     4081 ttttcgtgca tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt
     4141 tccttatatg tagctttcga catgatttat cttcgtttcc tgcaggtttt tgttctgtgc
     4201 agttgggtta agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat
     4261 accaatctaa gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca
     4321 aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaaa
     4381 aggtggtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4441 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4501 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4561 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4621 taataatggt ttcttagacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta
     4681 atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga
     4741 ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa
     4801 aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag
     4861 cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc
     4921 gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga
     4981 agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga
     5041 aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt
     5101 tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg
     5161 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5281 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5341 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5401 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5461 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5521 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5581 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5641 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5701 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5761 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5821 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5881 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5941 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6001 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6061 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6121 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6181 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6241 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6301 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6361 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6421 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6481 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6541 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6601 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6661 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6721 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6781 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6841 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6901 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6961 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7021 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7081 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7141 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca at
//
