LOCUS       sequence-246167-        9369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Repressor plasmid for WTC 846. Encodes TetR and TetR-Tup1, His3MX 
            selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9369)
  AUTHORS   Azizoglu A, Brent R, Rudolf F
  TITLE     A precisely adjustable, variation-suppressed eukaryotic 
            transcriptional controller to enable genetic discovery.
  JOURNAL   Elife. 2021 Aug 3;10. pii: 69549. doi: 10.7554/eLife.69549.
  PUBMED    34342575
REFERENCE   2  (bases 1 to 9369)
  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"; journalName: "Elife."; 
            date: "2021-08-3"; volume: "10. pii: 69549. doi"; pages: " 
            10.7554/eLife.69549"
COMMENT     Sequence Label: FRP2371
FEATURES             Location/Qualifiers
     source          1..9369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             814..834
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      877..1066
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1152..1170)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1191..1207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1191..1207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1204..1226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1215..1231
                     /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     complement(1465..1482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1636..2224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1716..1735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2403..3194,3195..3263))
                     /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     3026..3045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3264..3368)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3666..4316
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQK
                     GEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQA
                     MGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLYS
                     FSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     primer_bind     4655..4677
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4669..4686
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4670..4686
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4693..4712
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4693..4711
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5623..5642)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             6168..6188
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8500..8749
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
ORIGIN
        1 aaaaaaaagg ttgaaaccag ttccctgaaa ttattcccct atctatcatt gatagatata
       61 aatatctatc attgatagag taattctgta aatctatttc ttaaacttct taaattctac
      121 ttttatagtt agtctttttt ttagttttaa aacaccaaga acttagtttc gaataaacac
      181 acataaacaa acaaaatgtc caggttggac aagtccaagg ttattaactc tgccttggag
      241 ttgttgaacg aagttggtat tgaaggtttg accactagaa agttggctca aaaattgggt
      301 gttgaacaac ctaccttgta ctggcatgtt aagaacaaaa gagctttgtt ggacgctttg
      361 gccattgaaa tgttggatag acatcatacc catttctgtc cattggaagg tgaatcatgg
      421 caagatttct tgagaaacaa cgccaagtct ttcagatgtg ctttgttgtc tcatagagat
      481 ggtgctaaag ttcacttggg tactagacca actgaaaagc aatacgaaac tttggaaaac
      541 cagttggctt tcttgtgtca acaaggtttc tctttagaaa acgccttgta tgctttgtct
      601 gctgttggtc attttacctt gggttgtgtt ttggaagatc aagaacatca agtcgccaaa
      661 gaagaaaggg aaactccaac tactgattct atgccaccat tattgagaca agccattgag
      721 ttgtttgatc atcaaggtgc tgaaccagct tttttgtttg gtttggaatt gatcatctgc
      781 ggtttggaga aacaattgaa gtgtgaatct gcccctaaga agaagagaaa ggtttgaatt
      841 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
      901 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
      961 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     1021 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgggc
     1081 cgtatttaca gataacgatg tcgttattat atatatatat atatattgtc acttgtaccc
     1141 agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg
     1201 tttcctgtgt gaaattgtta tccgctcaca acagctgaca ccgattattt aaagctgcag
     1261 catacgatat atatacatgt gtatatatgt atacctatga atgtcagtaa gtatgtatac
     1321 gaacagtatg atactgaaga tgacaaggta atgcatcatt ctatacgtgt cattctgaac
     1381 gaggcgcgcc tctccctgca ggcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1441 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1501 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1561 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2041 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2101 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2221 tcaagaagat cctttgatct tttgtttaaa cctacggggt ctgacgctca gtggaacgaa
     2281 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2341 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2401 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2461 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2521 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2581 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2641 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2701 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2761 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2821 gcggttagct ccttcggtcc tccgatggtt gtcagaagta agttggccgc agtgttatca
     2881 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2941 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3001 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3061 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3121 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3181 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3241 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3301 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3361 gttccgcgca catttccccg aaaagtgcca cctgacgtcg aaagaacgcg gcattagcaa
     3421 catttgcgaa gcgaacgatt ttgcgaactg ctattaatta actctggcgc gcctcctcta
     3481 gtacactcta tattttttta tgcctcggta atgattttca tttttttttt tccacctagc
     3541 ggatgactct ttttttttct tagcgattgg cattatcaca taatgaatta tacattatat
     3601 aaagtaatgt gatttcttcg aagaatatac taaaaaatga gcaggcaaga taaacgaagg
     3661 cacatatgag gagggctttt gtagaaagaa atacgaacga aacgaaaatc agcgttgcca
     3721 tcgctttgga caaagctccc ttacctgaag agtcgaattt tattgatgaa cttataactt
     3781 ccaagcatgc aaaccaaaag ggagaacaag taatccaagt agacacggga attggattct
     3841 tggatcacat gtatcatgca ctggctaaac atgcaggctg gagcttacga ctttactcaa
     3901 gaggtgattt aatcatcgat gatcatcaca ctgcagaaga tactgctatt gcacttggta
     3961 ttgcattcaa gcaggctatg ggtaactttg ccggtgttaa aagatttgga catgcttatt
     4021 gtccacttga cgaagctctt tccagaagcg tagttgactt gtcgggacgg ccctatgctg
     4081 ttatcgattt gggattaaag cgtgaaaagg ttggggaatt gtcctgtgaa atgatccctc
     4141 acttactata ttccttttcg gtagctgctg gaattacttt gcatgttacc tgcttatatg
     4201 gtagtaatga ccatcatcgt gctgaaagcg cttttaaatc tctggctgtt gccatgcgcg
     4261 cggctacttc tcttactgga agttctgaag tcccaagcac gaagggagtg ttgtaaagat
     4321 gaattggatt atgtcaggaa aagaacgaca attttgcatc caaattgtct aaattttaga
     4381 gttgcttgaa aacaatagaa ccttacttgc tttataatta cgttaattag aagcgttatc
     4441 tcgtgaagga atatagtacg tagccgtata aattgaattg aatgttcagc ttatagaata
     4501 gagacacttt gctgttcaat gcgtcgtcac ttaccatact cactttatta tacgacttta
     4561 agtataaact ccgcggttat ggtaaaatta atgatgcaca aacgtccgat cggatgtgct
     4621 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     4681 gccagtgaat tgtaatacga ctcactatag ggcgaattgg agaaacagaa taatatataa
     4741 acctgtataa tataaccttg aagactatat ttcttttcgc ggccgcctat agggcgaatt
     4801 ggagctccac cgcggtggct ctagaagtcg aacaagaagc aggcaaagtt tagagcactg
     4861 cccctccgca ctcaaaaaag aaaaaactag gaggaaaata aaattctcaa ccacacaaac
     4921 acataaacac atacaaatac aaatacaagc ttatttactt gacatcgcgc gatcttccac
     4981 tattcagcgc cgtccgccct ctctcgtgtt ttttgtttac gcgacaacta tgcgaaatcc
     5041 ggagcaacgg gcaaccgttt ggggaaagac cacacccacg cgcgatcgcc atggcaacga
     5101 ggtcgcacac gccccacacc cagacctccc tgcgagcggg catgggtaca atgtccccgt
     5161 tgccacagag accacttcgt agcacagcgc agagcgtagc gtgttgttgc tgctgacaaa
     5221 agaaaatttt tcttagcaaa gcaaaggagg ggaagcacgg gcagatagca ccgtaccata
     5281 cccttggaaa ctcgaaatga acgaagcagg aaatgagaga atgagagttt tgtaggtata
     5341 tatagcggta gtgtttgcgc gttaccatca tcttctggat ctatctattg ttcttttcct
     5401 catcactttc ccctttttcg ctcttcttct tgtcttttat ttctttcttt tttttaattg
     5461 ttccctcgat tggctatcta ccaaagaatc caaacttaat acacgtattt atttgtccaa
     5521 ttaccggatc ccccgggctg caggaattca tgtccagatt agataaaagt aaagtgatta
     5581 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     5641 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     5701 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     5761 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     5821 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     5881 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     5941 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     6001 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     6061 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     6121 aattgatcat ttgcggatta gaaaaacaac ttaaatgtga aagtgccccc aagaaaaaga
     6181 gaaaggtggc agaagccgcc gcaaaggaag cggcagcaaa agaggctgcg gccaaggaag
     6241 ctgctgccaa ggcgctagaa gcagaggcgg ccgcaaagga ggcggccgcg aaggaagcgg
     6301 ctgcgaagga ggcagcggca aaggcgatga ctgccagcgt ttcgaatacg cagaataagc
     6361 tgaatgagct tctcgatgcc atcagacagg agtttctcca agtctcacaa gaggcaaata
     6421 cctaccgtct tcaaaaccaa aaggattacg atttcaaaat gaaccagcag ctggctgaga
     6481 tgcagcagat aagaaacacc gtctacgaac tggaactaac tcacaggaaa atgaaggacg
     6541 cgtacgaaga agagatcaag cacttgaaac tagggctgga gcaaagagac catcaaattg
     6601 catctttgac cgtccagcaa cagcggcaac agcaacagca gcaacaggtc cagcagcatt
     6661 tacaacagca acagcagcag ctagccgctg catctgcatc tgttccagtt gcgcaacaac
     6721 caccggctac tacttcggcc accgccactc cagcagcaaa cacaactact ggttcgccat
     6781 cggccttccc agtacaagct agccgtccta atctggttgg ctcacagttg cctaccacca
     6841 ctttgcctgt ggtgtcctca aacgcccaac aacaactacc acaacagcaa ctgcaacagc
     6901 agcaacttca acaacagcaa ccacctcccc aggtttccgt ggcaccattg agtaacacag
     6961 ccatcaacgg atctcctact tctaaagaga ccactacttt accctctgtc aaggcacctg
     7021 aatctacgtt gaaagaaact gaaccggaaa ataataatac ctcgaagata aatgacaccg
     7081 gatccgccac cacggccacc actaccaccg caactgaaac tgaaatcaaa cctaaggagg
     7141 aagacgccac cccggctagt ttgcaccagg atcactactt agtcccttat aatcaaagag
     7201 caaaccactc taaacctatc ccacctttcc ttttggatct agattcccag tctgttcccg
     7261 atgctctgaa gaagcaaaca aatgattatt atattttata caacccggca ctaccaagag
     7321 aaattgacgt tgagttacac aaatctttgg atcatacttc agttgtttgt tgcgtgaagt
     7381 tcagtaacga tggtgaatac ttagccacag gctgcaacaa aactactcaa gtgtatcgcg
     7441 tttcagatgg ttctctggtg gcccgtctat ctgacgattc tgctgccaat aaccatcgaa
     7501 attcgatcac tgaaaataac accaccacgt ccacggataa caatacaatg acaaccacta
     7561 ctaccaccac aattactacc acagcgatga cttcggcagc agaattggca aaagatgtgg
     7621 aaaacctgaa cacttcgtct tccccatcat ccgacttgta tatccgttca gtgtgttttt
     7681 ctccagatgg gaaatttttg gcaacaggtg ctgaagacag actgattaga atttgggata
     7741 ttgaaaatag aaagattgtt atgattcttc aaggccacga acaagatatt tattcattgg
     7801 actactttcc ctcaggtgac aaattagtct ccggttctgg tgaccgtacc gttcgtattt
     7861 gggacttacg tacaggccag tgttcattga ctttatccat tgaagatggt gttaccaccg
     7921 tcgctgtatc accaggtgat ggtaaataca tcgctgctgg ttctctagat cgtgctgtga
     7981 gagtttggga ttccgagacc ggattcttgg tggaaagact agattcggaa aacgaatccg
     8041 gtacaggcca caaggactct gtttatagcg ttgtcttcac tagagatgga caaagcgttg
     8101 tatccggctc attagataga tctgttaagc tctggaattt gcagaatgca aacaacaaga
     8161 gcgattcgaa aactccaaat tccggcactt gtgaagttac gtatatcggg cataaagact
     8221 ttgtattgtc cgtggccacc acacaaaatg atgagtacat cttgtccggt tccaaagatc
     8281 gtggtgtcct gttttgggat aagaaatccg gcaatccgtt attgatgttg caaggtcata
     8341 ggaattcagt tatatctgtg gctgtggcaa acgggtctcc gctgggtcca gaatataacg
     8401 tttttgctac tggtagcggt gattgtaaag caaggatttg gaagtataaa aaaatagcgc
     8461 caaattaact cgagacaggc cccttttcct ttgtctatat catgtaatta gttatgtcac
     8521 gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac
     8581 ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt
     8641 tatatttcaa atttttcttt tttttctgta caaacgcgtg tacgcatgta acattatact
     8701 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca aggttcgcag
     8761 tttacactct catcgtcgct ctcatcatcg cttccgttgt tgttttcctt agtagcgtct
     8821 gctttctaga cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat
     8881 aattaatagt agtgattttc ctaactttat ttagtcaaaa aattagcctc tatcattgat
     8941 agagtgtctg ggtgatctat cattgataga gcatccacta attttaattc tgctgtaacc
     9001 cgtacatgcc caaaataggg ggcgggttac acagaatcta tcattgatag agtgtctggg
     9061 tgatctatca ttgatagagc atccactaaa tataatggag ctctatcatt gatagagcat
     9121 ccactaaaaa aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat
     9181 aggtccattc tcttagcgca actacagaga acaggggcac aaacaggcaa aaaacgggca
     9241 caacctcaat ggagtgatgc aacctgcctg gagtaaatga tgacacaagg caattgaccc
     9301 acgcatgtat ctatctcatt ttcttacacc ttctattacc ttctgctctc tctgatttgg
     9361 aaaaagctg
//
