LOCUS       sequence-256132-        9625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reverse yTRAP, tetR-responsive Neon reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9625)
  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 9625)
  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: pSK221
FEATURES             Location/Qualifiers
     source          1..9625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3105..3123
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3126..3144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      3437..3446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5264..5283)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5265..5283)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5290..5307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5290..5306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5299..5321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5447..5902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5534..5553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5744..5765
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6258..6277)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6377..6399
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6437..6455)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6540..6644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6645..6713,6714..7505)
                     /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(6863..6882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7676..8264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8165..8184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8418..8435
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8552..8573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8588..8593,8594..8611,8612..8618)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8626..8642
                     /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     8631..8653
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8650..8666
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8650..8666
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8687..8705
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8993..9400
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
ORIGIN
        1 cgctcaaaag agcccggcac cggtaccggg caaaagcccc tagcaagcta aaatgattca
       61 actttcgtag taatttgttc aaccggtcac agtaaagtca atgactaact tttgaatcga
      121 ggatgtattt ccaattgaaa gcaatgtata tataaaccat tgtgttgaag cttgtcttac
      181 agttttcttg tatcctccca ttgttcatat agtacttaaa gtaccataat ccttaaagaa
      241 caaacttgaa agagaataca ctatacacaa ctaataaata aagaataatc atggctgtga
      301 ccaagacaat tgtagttcta ccaggtgacc atgttggtca agaaatcact gaagaggcca
      361 ttaaagtttt aaatgctatt caggaatgtc gtccagacaa ggtcaatttc aagtttgatc
      421 atcatttgat cggtggtgct gcaattgatg ccactggtgt tccattacca gacgaagctt
      481 tggaggcctc taagaaagct gatgccgtgc ttctgggtgc tgttggtggt ccaaaatggg
      541 gtactggcgc tgtcagacca gaacaaggtc ttttgaaaat ccgtaaggag ttgcaattgt
      601 atgctaatct aagaccatgt aattttgcat ctgattcctt actagatcta tcgccattga
      661 agcctgaaat tgcaagaggt acagatttcg tcgttgttag agaactagtg ggtggtattt
      721 atttcgggga gagaaaagaa gatgaaggtg atggtgtcgc ctgggatagc gaaaagtatt
      781 ctgtgcccga agttcaaaga atcactagaa tggctgcttt catggctctg caacacaacc
      841 cacctttacc tatttggtct ctagataaag ccaacgttct agcatcatca agattatgga
      901 gaaaaacagt ggaagagacg attaagaatg aattcccaca attgactgtt caacatcaat
      961 tgattgattc cgctgctatg atcttggtta agaacccaac tcatctaaat ggtattatta
     1021 tcacaaacaa tatgtttggt gatattatct ccgatgaagc ctccgtgatc ccaggttcgt
     1081 taggattact tccatctgcc tccttggcat ctcttccaga taagaatacg gcatttggtt
     1141 tatatgagcc atgtcatggc tctgctccag acttaccaaa gggaaaggtt aaccccgtag
     1201 caacaatcct atctgctgct atgatgttga agttgtctct ggacctgttc gaagaaggtg
     1261 aaattataga acaagcagtc aagaaagtgt tggattctgg tatcagaaca gcggatctaa
     1321 agggtaccaa ctctactact gaggtcggtg atgcagtagc gaaagctgtc agggaactat
     1381 tagcttagac ttagtagatg tacagttgca tacataagtc tgaacagaaa gaagtaaata
     1441 tcatatctag ttcggtttag aaatcagtga ctatattggt tttgaccaga tcgcaatcct
     1501 tgcaagcatt tagtaatgac tatcaaaccg gtgaatcatc tgataaggct atattagctt
     1561 gtgcattcgc atgtatcggg aaacgaacgg taccgggccc ataactcagc cctcgaggga
     1621 ggtcttgggg agcaaggcag gtgttacggc gataaagctt caagttacag aattcttact
     1681 tcagctgcag aagaaatgat ggtaaatgaa ataggaaatc aaggagcatg aaggcaaaag
     1741 acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt tgatatgatg tatttggctt
     1801 tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca tgctgactct gtggcggacc
     1861 cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc tgtgcccggc ggagtttttt
     1921 gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg agattggaga agcaataaga
     1981 atgccggttg gggttgcgat gatgacgacc acgacaactg gtgtcattat ttaagttgcc
     2041 gaaagaacct gagtgcattt gcaacatgag tatactagaa gaatgagcca agacttgcga
     2101 gacgcgagtt tgccggtggt gcgaacaata gagcgaccat gaccttgaag gtgagacgcg
     2161 cataaccgct agagtacttt gaagaggaaa cagcaatagg gttgctacca gtataaatag
     2221 acaggtacat acaacactgg aaatggttgt ctgtttgagt acgctttcaa ttcatttggg
     2281 tgtgcacttt attatgttac aatatggaag ggaactttac acttctccta tgcacatata
     2341 ttaattaaag tccaatgcta gtagagaagg ggggtaacac ccctccgcgc tcttttccga
     2401 tttttttcta aaccgtggaa tatttcggat atccttttgt tgtttccggg tgtacaatat
     2461 ggacttcctc ttttctggca accaaaccca tacatcggga ttcctataat accttcgttg
     2521 gtctccctaa catgtaggtg gcggagggga gatatacaat agaacagata ccagacaaga
     2581 cataatgggc taaacaagac tacaccaatt acactgcctc attgatggtg gtacataacg
     2641 aactaatact gtagccctag acttgatagc catcatcata tcgaagtttc actacccttt
     2701 ttccatttgc catctattga agtaataata ggcgcatgca acttcttttc tttttttttc
     2761 ttttctctct cccccgttgt tgtctcacca tatccgcaat gacaaaaaaa tgatggaaga
     2821 cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg
     2881 atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct
     2941 ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaaagt
     3001 ttgctgtctt gctatcaagt ataaatagac ctgcaattat taatcttttg tttcctcgtc
     3061 attgttctcg ttccctttct tccttgtttc tttttctgca caattcccta tcagtgatag
     3121 agaattccct atcagtgata gagagctagc ttcaagctat accaagcata caatcaactc
     3181 ccgggaaaaa atgggcgagg aggataacat ggcctctctc ccagcgacac atgagttaca
     3241 catctttggc tccatcaacg gtgtggactt tgacatggtg ggtcagggca ccggcaatcc
     3301 aaatgatggt tatgaggagt taaacctgaa gtccaccaag ggtgacctcc agttctcccc
     3361 ctggattctg gtccctcata tcgggtatgg cttccatcag tacctgccct accctgacgg
     3421 gatgtcgcct ttccaggccg ccatggtaga tggctccgga taccaagtcc atcgcacaat
     3481 gcagtttgaa gatggtgcct cccttactgt taactaccgc tacacctacg agggaagcca
     3541 catcaaagga gaggcccagg tgaaggggac tggtttccct gctgacggtc ctgtgatgac
     3601 caactcgctg accgctgcgg actggtgcag gtcgaagaag acttacccca acgacaaaac
     3661 catcatcagt acctttaagt ggagttacac cactggaaat ggcaagcgct accggagcac
     3721 tgcgcggacc acctacacct ttgccaagcc aatggcggct aactatctga agaaccagcc
     3781 gatgtacgtg ttccgtaaga cggagctcaa gcactccaag accgagctca acttcaagga
     3841 gtggcaaaag gcctttaccg atgtgatggg catggacgag ctgtacaagt aaggatccgt
     3901 cccaattact agtgtttatc catctagagt accctaccac ctgcaacatg cgtacgaaga
     3961 cagcggccgc gattcgaatc accgcggtgt cataacttgt gagctctaag caaatagcta
     4021 aattatatac gaattaatat tatgattaag tgtttacgtg agtgcgatat ttttattact
     4081 atcttataca gttgtatata ctctataaaa tgagttgtct attaattaac gcgatgaatg
     4141 ctttctgggt ttacctctcc aacaactcta gtttacttct caatacattc aattgtattt
     4201 gatttgtcaa tacttcatca ttaatcaatt ctatagtttt gtttttctcg tttatttcca
     4261 aatttaatgc atcaatttta ttattcaatt tgtcgttgat tttggttaat gattttatgg
     4321 tttgatctct ggcattgatt gtttgtgtta gtttttcatt attgataatt aaattattta
     4381 agttagttat caactcggtg ttttcaagtt tcaagttttc aatttcttta gagtttatta
     4441 gatttgtcaa agtttctgaa ttgcttgatt ggtcctgtag aagagtattt gttgttgtgg
     4501 ataattgatt caatttttga gacaattgct ggaaggcgtt gaaatatcta gcatcaatct
     4561 catggttttt ttcccgagag tctcgtagat tcaattgttt taatatatct tgggaccact
     4621 cttgatttga actcatggaa attaaactgg gtgttgtgtt gtggtgtaat gattgtaccc
     4681 cctttgctta taattgtgtg gccaccgaag tcggtgatgc tgtcgccgaa gaagttaaga
     4741 aaatccttgc ttaaaaagat tctctttttt tatgatattt gtacataaac tttataaatg
     4801 aaattcataa tagaaacgac acgaaattac aaaatggaat atgttcatag ggtagacgaa
     4861 actatatacg caatctacat acatttatca agaaggagaa aaaggaggat gtaaaggaat
     4921 acaggtaagc aaattgatac taatggctca acgtgataag gaaaaagaat tgcactttaa
     4981 cattaatatt gacaaggagg agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg
     5041 aaactatgat tcctaattta tatattggag gattttctct aaaaaaaaaa aaatacaaca
     5101 aataaaaaac actcaatgac ctgaccattt gatggagttt aagtcaatac ctccttgaac
     5161 catttcccat aatggtgaaa gttccctcaa gaattttact ctgtcagaaa cggccttaac
     5221 gacgtaggcc ggccatttaa atgtttaaac ggccacgctg gccccctata gtgagtcgta
     5281 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     5341 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     5401 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     5461 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5521 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5581 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5641 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5701 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5761 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5821 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5881 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5941 tttcacaccg catagggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     6001 agtattagga tctctaccct tggcgaaaag tcctctgcca acaatgatga tatctgatcc
     6061 accacttaca acttcgtcga cggttctgta ctgctgaccc aatatgcgta tatataccaa
     6121 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     6181 atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg
     6241 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     6301 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     6361 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     6421 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6481 atggtttctt agacgtcggc gcgccgagga cgtcaggtgg cacttttcgg ggaaatgtgc
     6541 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6601 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6661 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6721 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6781 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6841 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6901 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6961 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7021 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7081 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7141 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7201 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7261 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7321 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7381 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7441 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7501 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7561 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7621 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7681 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7741 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7801 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7861 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7921 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7981 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8041 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8101 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8161 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8221 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8281 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8341 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8401 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     8461 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     8521 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     8581 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     8641 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt aaccctcact
     8701 aaagggaaca aaagctgggc cggccattta aatgtttaaa cggccacgct ggccgaggag
     8761 aacttctagt atatctacat acctaatatt attgccttat taaaaatgga atcccaacga
     8821 ttacatcaaa atccacattc tcttcaaaat caattgtcct gtacttcctt gttcatgtgt
     8881 gttcaaaaac gttatattta taggataatt atactctatt tctcaacaag taattggttg
     8941 tttggccgag cggtctaagg cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg
     9001 gaatactcag gtatcgtaag atgcaagagt tcgaatctct tagcaaccat tatttttttc
     9061 ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg atgactggaa
     9121 attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg aaaaattggg
     9181 agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga gaagcgttca
     9241 tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg acctattgtt
     9301 ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct tttacatttc
     9361 agcaatatat atatatatat ttcaaggata taccattcta atgtctgccc ctaagttacc
     9421 aatagatagc ggtttaatat aaggtttcaa caagaaatat tttgtgaaaa tcaaaagtta
     9481 atagtttcac tgtcgaaaat atgaaagttt aagttttcag ttcatcccat tgtagttatc
     9541 aaatttctgt gtttcccgta tatgtgtgct ctgtattcat cgctatcttg aaactggtta
     9601 tgatcgttgc gcatgtgacc tgtat
//
