LOCUS       sequence-189769-        7902 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 7902)
  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 7902)
  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-NQYFP
FEATURES             Location/Qualifiers
     source          1..7902
                     /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(4116..4778)
                     /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(4779..4966)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(5113..5132)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5232..5254
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5292..5310)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5358..5861
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5891..5995
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5996..6064,6065..6856)
                     /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(6214..6233)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7027..7615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7516..7535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7769..7786
                     /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 accgcataga tccgtcgagt tcaagagaaa aaaaaagaaa aagcaaaaag aaaaaaggaa
     3961 agcgcgcctc gttcagaatg acacgtatag aatgatgcat taccttgtca tcttcagtat
     4021 catactgttc gtatacatac ttactgacat tcataggtat acatatatac acatgtatat
     4081 atatcgtatg ctgcagcttt aaataatcgg tgtcactaca taagaacacc tttggtggag
     4141 ggaacatcgt tggtaccatt gggcgaggtg gcttctctta tggcaaccgc aagagccttg
     4201 aacgcactct cactacggtg atgatcattc ttgcctcgca gacaatcaac gtggagggta
     4261 attctgctag cctctgcaaa gctttcaaga aaatgcggga tcatctcgca agagagatct
     4321 cctactttct ccctttgcaa accaagttcg acaactgcgt acggcctgtt cgaaagatct
     4381 accaccgctc tggaaagtgc ctcatccaaa ggcgcaaatc ctgatccaaa cctttttact
     4441 ccacgcacgg cccctagggc ctctttaaaa gcttgaccga gagcaatccc gcagtcttca
     4501 gtggtgtgat ggtcgtctat gtgtaagtca ccaatgcact caacgattag cgaccagccg
     4561 gaatgcttgg ccagagcatg tatcatatgg tccagaaacc ctatacctgt gtggacgtta
     4621 atcacttgcg attgtgtggc ctgttctgct actgcttctg cctctttttc tgggaagatc
     4681 gagtgctcta tcgctagggg accacccttt aaagagatcg caatctgaat cttggtttca
     4741 tttgtaatac gctttactag ggctttctgc tctgtcatct ttgccttcgt ttatcttgcc
     4801 tgctcatttt ttagtatatt cttcgaagaa atcacattac tttatataat gtataattca
     4861 ttatgtgata atgccaatcg ctaagaaaaa aaaagagtca tccgctaggt ggaaaaaaaa
     4921 aaatgaaaat cattaccgag gcataaaaaa atatagagtg tactagagga ggccaagagt
     4981 aatagaaaaa gaaaattgcg ggaaaggact gtgttatgac ttccctgact aatgccgtgt
     5041 tcaaacgata cctggcagtg actcctagcg ctcaccaagc tcttaaaacg ggaatttatg
     5101 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5161 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5221 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5281 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5341 ttcttagggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat
     5401 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
     5461 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
     5521 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
     5581 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
     5641 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
     5701 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
     5761 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
     5821 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtcaggtg gcacttttcg
     5881 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5941 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6001 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6061 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6121 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6181 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6241 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6301 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6361 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6421 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6481 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6541 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6601 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6661 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6721 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6781 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6841 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6901 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6961 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7021 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7081 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7141 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7201 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7261 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7321 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7381 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7441 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7501 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7561 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7621 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7681 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7741 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     7801 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7861 caggtttccc gactggaaag cgggcagtga gcgcaacgca at
//
