LOCUS       sequence-255863-        8942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ""ZF only"" yTRAP control to control for yTRAP reporter fluorescence
            changes not dependent on sensed protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8942)
  AUTHORS   Newby GA, Kiriakov S, Hallacli E, Kayatekin C, Tsvetkov P, Mancuso 
            CP, Bonner JM, Hesse WR, Chakrabortee S, Manogaran AL, Liebman SW, 
            Lindquist S, Khalil AS
  TITLE     A Genetic Tool to Track Protein Aggregates and Control Prion 
            Inheritance.
  JOURNAL   Cell. 2017 Nov 2;171(4):966-979.e18. doi: 
            10.1016/j.cell.2017.09.041. Epub 2017 Oct 19.
  PUBMED    29056345
REFERENCE   2  (bases 1 to 8942)
  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: 
            "10.1016/j.cell.2017.09.041"; journalName: "Cell"; date: "2017-11-2-
            2"; volume: "171"; issue: "4"; pages: "966-979.e18"
COMMENT     Sequence Label: pSK267
FEATURES             Location/Qualifiers
     source          1..8942
                     /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."
     protein_bind    77..93
                     /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     82..104
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     101..117
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     101..117
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1609..1629
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2233..2322
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             2329..2418
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     regulatory      3555..3564
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      4031..4218
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4086..4110)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            4293..5424
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     CDS             4643..5215
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      5227..5424
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     5987..6003
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5987..6003
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6017..6036)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6018..6036)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6043..6060)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6043..6059)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6052..6074)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6359..6378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6491..6512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6666..6685)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6785..6807
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6845..6863)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6931..7035
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7036..7104,7105..7896)
                     /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(7254..7273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8067..8655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8556..8575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8809..8826
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccgcggct
       61 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      121 gattacgcca ctagtccgag gcctcgagat ccgatatcgc cgtggcggcc gcaattatcc
      181 tgggcacgag tgaaacaaag ctaaaacctt tatttagcat ggccattgaa tgtaacaatt
      241 atatatatcg caagcacaaa aaatcaagga gagagaacta ccactttgtt catgtgtaca
      301 atgttcatta tctccataag caaaaaaaaa aaaatagaaa acatatgcta taaggttgat
      361 attctcacga gtaagcggca cttgctactt attgacattg cagatttttg gctacagaaa
      421 tagtatatta gagattataa ttgctaatca aatcaaaata taaaattagt aaaccaaacc
      481 atttataccc ttccttagta gttatggatt gttttttaat gatatttctg caaaccaaag
      541 aaagattgtt atccagatag aatttagttt tgatattcat ttttttgttg aagattgaac
      601 gccatatctg ggcctcataa ttcaaaagac ggtgccatta tcggtagcgt ttcgcattgt
      661 actggatttc agaaatttca cagttgatga atcgaaaaga atggtctcat tgcaacacgt
      721 aaggttaaga tgtccctttt taccattata ggcaataaat gaatcataaa acgaccgtat
      781 actggtgaaa tagtagggag aacgagtacc tgtagtaaaa agtataaatc atagttaatc
      841 gggcaatgtc cctcgatcaa ggagtattgt gtcatgttcg agacaaacgc caacattttt
      901 gtttcttttg gacaaatgtt gtttgcattt atgatccgtt atattttgat ctaatgtaga
      961 gttgcacgta gttcttactg gcaaagaaat cgatgcatac caaaaaagaa taaaggtgat
     1021 atttgatctt taccgtttag ttccaacgta aaattgtgcc tttggactta aaatggcgtc
     1081 gtacgctgca ccgagtgcaa ccacacaaaa atcatacaac gaatggtata tgcttcattt
     1141 ctttgtttcg cattagctgc gctatttgac tcaaattatt attttttact aagacgacgc
     1201 gtcacagtgt tcgagtctgt gtcatttctt ttgtaattct cttaaaccac ttcataaagt
     1261 tgtgaagttc atagcaaaat tcttccgcaa aaagatgaat cttagttctc agcccaccaa
     1321 aagaggtaca tgctaagatc atacagaagt tattgtcact tcttaccttg ctcttaaatg
     1381 tacattacaa ccgggtatta tatcttacat catcgtataa tatgatcttt ctttatggag
     1441 aaaatttttt tttcactcga ccaaagctcc cattgcttct gaagagtgta gtgtatattg
     1501 gtacatcttc tcttgaaaga ctccattgta ctgtaacaaa aagcggtttc ttcatcgact
     1561 tgctcggaat aacatctata tctgcccact agcaacagct taatggctcc aaagaagaag
     1621 agaaaggttg gtattcatgg tgttccagtt gatggtggtt tacaagctcc accaactgat
     1681 gtttctttgg gtgatgaatt gcatttggat ggtgaagatg ttgctatggc tcatgctgat
     1741 gctttggatg attttgattt ggatatgttg ggtgacggtg attctccagg tccaggtttt
     1801 actccacatg attctgctcc atatggtgct ttagatatgg ctgactttga attcgaacaa
     1861 atgttcaccg atgccttggg tattgatgaa tatggtggtg gtgctagacc aggtgaaaga
     1921 ccatttcaat gtagaatctg catgagaaac ttctccagac aagatagatt ggatagacat
     1981 accagaactc atactggtga aaagccattc caatgcagaa tatgtatgag aaatttttcc
     2041 caaaaagaac acttggccgg tcatttgaga acacacacag gtgaaaaacc ttttcaatgc
     2101 agaatatgca tgagaaactt tagtagaaga gacaacttga acagacactt gaaaacccat
     2161 ttgagaggtt ctgccggcga attggctttg aaattggctg gtttggatat caattctggt
     2221 tctgctgcta gatacccata tgatgttcca gattatgctg gttaccctta cgatgttcct
     2281 gactacgcag gttcttatcc atacgatgta cctgattatg cctcaagata tccttatgac
     2341 gtccctgatt acgccggtta tccatatgac gtaccagact acgctggtag ttacccatac
     2401 gacgtcccag attacgcttc ttcataatga gggccctcat gtaatttctt gcaaacataa
     2461 gtaaatgcaa acacaataat accgatcata aagcattttc ttctatatta aaaaacaagg
     2521 tttaataaag ctgttatata tatatatata tatatagacg tataattagt ttagttcttt
     2581 ttgtaccata taccataaac aaggtaaact tcacctctca atatatctag aatttcataa
     2641 aaatatctag caaggtttca actccttcaa tcacgttttc atcactcaac cctatctcgg
     2701 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2761 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcctgat
     2821 gcggtatttt ctttaattaa agagtgagga ctcgaaaata ttaatagagt gaggactcga
     2881 aaatattaat agagtgagga ctcgaaaata ttaatagagt gaggactcga aaatattaat
     2941 atcgatagag tgaggactcg aaaatattaa tagagtgagg actcgaaaat attaatgaat
     3001 tcagagtgag gactcgaaaa tattaataga gtgaggactc gaaaatatta atggccggcc
     3061 cagatccgcc aggcgtgtat atatagcgtg gatggccagg caactttagt gctgacacat
     3121 acaggcatat atatatgtgt gcgacgacac atgatcatat ggcatgcatg tgctctgtat
     3181 gtatataaaa ctcttgtttt cttcttttct ctaaatattc tttccttata cattaggacc
     3241 tttgcagcat aaattactat acttctatag acacacaaac acaaatacac acactgttta
     3301 aacaaaaaat gggcgaggag gataacatgg cctctctccc agcgacacat gagttacaca
     3361 tctttggctc catcaacggt gtggactttg acatggtggg tcagggcacc ggcaatccaa
     3421 atgatggtta tgaggagtta aacctgaagt ccaccaaggg tgacctccag ttctccccct
     3481 ggattctggt ccctcatatc gggtatggct tccatcagta cctgccctac cctgacggga
     3541 tgtcgccttt ccaggccgcc atggtagatg gctccggata ccaagtccat cgcacaatgc
     3601 agtttgaaga tggtgcctcc cttactgtta actaccgcta cacctacgag ggaagccaca
     3661 tcaaaggaga ggcccaggtg aaggggactg gtttccctgc tgacggtcct gtgatgacca
     3721 actcgctgac cgctgcggac tggtgcaggt cgaagaagac ttaccccaac gacaaaacca
     3781 tcatcagtac ctttaagtgg agttacacca ctggaaatgg caagcgctac cggagcactg
     3841 cgcggaccac ctacaccttt gccaagccaa tggcggctaa ctatctgaag aaccagccga
     3901 tgtacgtgtt ccgtaagacg gagctcaagc actccaagac cgagctcaac ttcaaggagt
     3961 ggcaaaaggc ctttaccgat gtgatgggca tggacgagct gtacaagtaa ggcgcgccac
     4021 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     4081 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
     4141 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
     4201 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc
     4261 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
     4321 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
     4381 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
     4441 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
     4501 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
     4561 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
     4621 ccgaacatca attgaaacaa ccatgggtac cactcttgac gacacggctt accggtaccg
     4681 caccagtgtc ccgggggacg ccgaggccat cgaggcactg gatgggtcct tcaccaccga
     4741 caccgtcttc cgcgtcaccg ccaccgggga cggcttcacc ctgcgggagg tgccggtgga
     4801 cccgcccctg accaaggtgt tccccgacga cgaatcggac gacgaatcgg acgacgggga
     4861 ggacggcgac ccggactccc ggacgttcgt cgcgtacggg gacgacggcg acctggcggg
     4921 cttcgtggtc gtctcgtact ccggctggaa ccgccggctg accgtcgagg acatcgaggt
     4981 cgccccggag caccgggggc acggggtcgg gcgcgcgttg atggggctcg cgacggagtt
     5041 cgcccgcgag cggggcgccg ggcacctctg gctggaggtc accaacgtca acgcaccggc
     5101 gatccacgcg taccggcgga tggggttcac cctctgcggc ctggacaccg ccctgtacga
     5161 cggcaccgcc tcggacggcg agcaggcgct ctacatgagc atgccctgcc cctaaagatc
     5221 ttcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     5281 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     5341 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     5401 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg
     5461 aaaacgagct cgaattcctg ggggaacaac ttcacagaat gttttgtcat attgtcgaag
     5521 tggtcacaaa acaagagaag ttccgccaat tataaaaagg gaacccgtat atttcagctt
     5581 cacggatgat ttccagggtg agagtactgt atatgggctt acgatagaag gccataaaaa
     5641 tttcttgctt ggcaacaaaa tagaagtgaa atcatgtcga ggctgctgtg tgggagaaca
     5701 gcataaaata tcacaaaaaa agaatctaaa acactgtgtt gcttgtccca gaaagggaat
     5761 caagtatttt tataaagatt ggagtggtaa aaatcgagta tgtgctagat gctatggaag
     5821 atacaaattc agcggtcatc actgtataaa ttgcaagtat gtaccagaag cacgtgaagt
     5881 gaaaaaggca aaagacaaag gcgaaaaatt gggcattacg cccgaaggtt tgccagttaa
     5941 aggaccagag tgtataaaat gtggcggaat cttacagtgc ggccgctcta gaactagtgg
     6001 atcgatcccc aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca
     6061 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     6121 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     6181 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     6241 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     6301 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     6361 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6421 aatcggggtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6481 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     6541 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     6601 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     6661 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     6721 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6781 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     6841 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6901 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6961 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     7021 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7081 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7141 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     7201 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7261 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7321 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7381 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7441 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7501 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7561 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7621 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7681 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7741 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7801 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7861 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7921 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7981 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     8041 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     8101 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     8161 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     8221 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     8281 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     8341 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8401 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8461 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8521 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8581 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8641 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8701 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     8761 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     8821 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     8881 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     8941 at
//
