LOCUS       sequence-189333-        7275 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 7275)
  AUTHORS   Du Z, Goncharoff DK, Cheng X, Li L
  TITLE     Analysis of [SWI+ ] formation and propagation events.
  JOURNAL   Mol Microbiol. 2017 Apr;104(1):105-124. doi: 10.1111/mmi.13616. Epub
            2017 Jan 26.
  PUBMED    28035761
REFERENCE   2  (bases 1 to 7275)
  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: "10.1111/mmi.13616";
            journalName: "Mol Microbiol"; date: "2017-04"; volume: "104"; issue:
            "1"; pages: "105-124"
COMMENT     Sequence Label: p413TEF-NYFP
FEATURES             Location/Qualifiers
     source          1..7275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    28..49
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(64..69,70..87,88..94)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    102..118
                     /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     107..129
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     126..142
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     126..142
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     161..181
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        163..181
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        199..597
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     regulatory      1612..1621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1618..1620,1621..1623,1624..2337)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1663..1684)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1924..1943)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2271..2292
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2350..2597
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2366..2384)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2615..2634)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2616..2634)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2644..2661)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2644..2660)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2653..2675)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2801..3256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2888..2907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3098..3119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3516..4178)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(4179..4366)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(4513..4532)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4632..4654
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4692..4710)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4758..5261
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5291..5395
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5396..5464,5465..6256)
                     /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(5614..5633)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6427..7015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6916..6935
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7169..7186
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
       61 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      121 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
      181 ggaacaaaag ctggagctca tagcttcaaa atgtttctac tcctttttta ctcttccaga
      241 ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa
      301 tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga
      361 aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac
      421 gtttcttttt cttgaaaatt ttttttttga tttttttctc tttcgatgac ctcccattga
      481 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      541 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      601 tctagaacta gtatggattt ctttaatttg aataataata ataataataa taatactact
      661 actactacta ctactaccaa taacaataat actaataata ataatactaa taataataat
      721 aatcctgcta ataataccaa taacaacaat agcacaggcc atagtagtaa tactaataac
      781 aatactaata acaacaatac gaataccgga gccagtggtg tagacgattt tcagaatttt
      841 ttcgacccaa aaccttttga ccagaatttg gattccaaca ataacaacag taatagtaat
      901 aacaatgata acaataacag taacacggta gcctccagca cgaactttac ttctccaaca
      961 gcagtcgtta ataatgccgc tcctgctaat gttaccggtg gaaaagctgc taatttcatt
     1021 cagaaccagt ctccccaatt caattctcca tatgactcaa acaattccaa cacaaattta
     1081 aacagcctgt ctcctcaggc tattttagcc aagaattcaa tcattgattc ttccaattta
     1141 cctcttcaag ctcaacaaca gctatatggc ggtaacaata ataataatag caccggaatt
     1201 gcgaacgata atgtaataac accccatttt atcaccaatg ttcaatccat cagtcaaaat
     1261 tcctcatctt ctactccgaa cacaaactct aactctactc cgaatgcaaa tcaacaattt
     1321 ttgccattca ataacagtgc ttccaataat ggtaatttga cgtctaacca gcttatttct
     1381 aattacgctg cttcaaattc aatggacaga tcatcctctg caagcaatga atttgttccg
     1441 aacacaagtg acaacaacaa caacagtaat aaccataata tgcgtaataa cagcaataat
     1501 aaaacgagca ataacaataa tgtcactgcc gtgcctgctg caactcccgc taataccaat
     1561 aattcaactt caaacgcaaa tacagtgttc tctgcccggg atccaccggt cgccaccatg
     1621 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1681 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1741 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1801 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     1861 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1921 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1981 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2041 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2101 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2161 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2221 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2281 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     2341 ggcctcgagt catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     2401 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     2461 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     2521 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     2581 cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     2641 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     2701 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     2761 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
     2821 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2881 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2941 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3001 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3061 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3121 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3181 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3241 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3301 accgcataga tccgtcgagt tcaagagaaa aaaaaagaaa aagcaaaaag aaaaaaggaa
     3361 agcgcgcctc gttcagaatg acacgtatag aatgatgcat taccttgtca tcttcagtat
     3421 catactgttc gtatacatac ttactgacat tcataggtat acatatatac acatgtatat
     3481 atatcgtatg ctgcagcttt aaataatcgg tgtcactaca taagaacacc tttggtggag
     3541 ggaacatcgt tggtaccatt gggcgaggtg gcttctctta tggcaaccgc aagagccttg
     3601 aacgcactct cactacggtg atgatcattc ttgcctcgca gacaatcaac gtggagggta
     3661 attctgctag cctctgcaaa gctttcaaga aaatgcggga tcatctcgca agagagatct
     3721 cctactttct ccctttgcaa accaagttcg acaactgcgt acggcctgtt cgaaagatct
     3781 accaccgctc tggaaagtgc ctcatccaaa ggcgcaaatc ctgatccaaa cctttttact
     3841 ccacgcacgg cccctagggc ctctttaaaa gcttgaccga gagcaatccc gcagtcttca
     3901 gtggtgtgat ggtcgtctat gtgtaagtca ccaatgcact caacgattag cgaccagccg
     3961 gaatgcttgg ccagagcatg tatcatatgg tccagaaacc ctatacctgt gtggacgtta
     4021 atcacttgcg attgtgtggc ctgttctgct actgcttctg cctctttttc tgggaagatc
     4081 gagtgctcta tcgctagggg accacccttt aaagagatcg caatctgaat cttggtttca
     4141 tttgtaatac gctttactag ggctttctgc tctgtcatct ttgccttcgt ttatcttgcc
     4201 tgctcatttt ttagtatatt cttcgaagaa atcacattac tttatataat gtataattca
     4261 ttatgtgata atgccaatcg ctaagaaaaa aaaagagtca tccgctaggt ggaaaaaaaa
     4321 aaatgaaaat cattaccgag gcataaaaaa atatagagtg tactagagga ggccaagagt
     4381 aatagaaaaa gaaaattgcg ggaaaggact gtgttatgac ttccctgact aatgccgtgt
     4441 tcaaacgata cctggcagtg actcctagcg ctcaccaagc tcttaaaacg ggaatttatg
     4501 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4561 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4621 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4681 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4741 ttcttagggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat
     4801 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
     4861 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
     4921 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
     4981 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
     5041 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
     5101 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
     5161 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
     5221 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtcaggtg gcacttttcg
     5281 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5341 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5401 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5461 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5521 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5581 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5641 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5701 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5761 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5821 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5881 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5941 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6001 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6061 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6121 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6181 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6241 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6301 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6361 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6541 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6781 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6841 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6901 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6961 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7021 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7081 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7141 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     7201 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7261 caggtttccc gactg
//
