LOCUS       sequence-246160-        9970 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Repressor plasmid for WTC 846. Encodes TetR and TetR-Tup1, Ura3 
            selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9970)
  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 9970)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: FRP2365
FEATURES             Location/Qualifiers
     source          1..9970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(340..537)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(1356..1699)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     1787..1809
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1801..1818
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1802..1818
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1825..1844
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1825..1843
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2755..2774)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             3300..3320
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5632..5881
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             7315..7335
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7378..7567
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(7653..7671)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7692..7708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7692..7708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7705..7727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7716..7732
                     /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(8042..8059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8213..8801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8293..8312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8980..9771,9772..9840))
                     /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     9603..9622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9841..9945)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaact ctggcgcgcc actacttttt ggaactcttg ttgttctttg gagttcaatg
      121 cgtccatctt tacagtcctg tcttattgtt cttgatttgt gccccgtaaa atactgttac
      181 ttggttctgg cgaggtattg gatagttcct ttttataaag gccatgaagg tttttctttc
      241 caattttttt tttttcgtca ttatagaaat cattacgacc gagattcccg gcatatgcgt
      301 tttcgacact ggatggcggc gttagtatcg aatcgacagc agtatagcga ccagcattca
      361 catacgattg acgcatgata ttactttctg cgcacttaac ttcgcatctg ggcagatgat
      421 gtcgaggcga aaaaaaatat aaatcacgct aacatttgat taaaatagaa caactacaat
      481 ataaaaaaac tatacaaatg acaagttctt gaaaacaaga atctttttat tgtcagtact
      541 gattataatt ggccagtctt tttcaaataa gcattccaac cagcatctct atacctttta
      601 ccttcaatat ctggatctct tcctttacca aacaatcctc taccaacaat gataatatca
      661 gttccagtgc taacaacttc atcaacagtt ctatattgtt gtcctaatcc atcaccttta
      721 tcatctaatc caactccagg tgtcataata agccaatcaa atccttcttc ttgtccaccc
      781 atatcacgtt gggcaataaa tccaataaca aattccttat cggatttagc aatttcaaca
      841 gttttttgag aatattctcc ataagctaat gatcccactg atgataattc agctaacatc
      901 aataaccctc ttggctcttg gttggtggtg gtttctttag ctccctgttt taatccttca
      961 actactccat tcccagtgac accatgagca ttagtaatat ctgcccaact actaatttta
     1021 taaactccac caatatattg tttcttcacg gtattaccaa tatcagcaaa ttttctatct
     1081 tcaaaaatca taaattgatg tttacgtgaa agttctaata atggttcaat agtggattca
     1141 taggaaaaat cattgattat atcaatatga gtcttgatta agcatacata aggacccaat
     1201 ttatcaatta attcaagaaa ttccttagtg gtatcaacat caattgatgc acataaattg
     1261 gttttcttca gttccattaa tcgaaataat cgttgtgcta ctggtgaggc atgagtttct
     1321 gctctctcac tataggtctt agtgttgact gtcatggttg tttatgttcg gatgtgatgt
     1381 gagaactgta tcctagcaag attttaaaag gaagtatatg aaagaagaac ctcagtggca
     1441 aatcctaacc ttttatattt ctctacaggg gcgcggcgtg gggacaattc aacgcgtctg
     1501 tgaggggagc gtttccctgc tcgcaggtcc gcagcgagga gccgtaattt ttgcttcgcg
     1561 ccgtgcggcc atcaaaatgt atggatgcaa atgattatac atggggatgt atgggctaaa
     1621 tgtacgggcg acagtcacat catgcccctg agctgcgcac gtcaagactg tcaaggaggg
     1681 tattctgggc ctccatgtcg ctggccgggt gacccggcgg ggacgaggca agctaaaccg
     1741 atcggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     1801 tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt ggagaaacag
     1861 aataatatat aaacctgtat aatataacct tgaagactat atttcttttc gcggccgcct
     1921 atagggcgaa ttggagctcc accgcggtgg ctctagaagt cgaacaagaa gcaggcaaag
     1981 tttagagcac tgcccctccg cactcaaaaa agaaaaaact aggaggaaaa taaaattctc
     2041 aaccacacaa acacataaac acatacaaat acaaatacaa gcttatttac ttgacatcgc
     2101 gcgatcttcc actattcagc gccgtccgcc ctctctcgtg ttttttgttt acgcgacaac
     2161 tatgcgaaat ccggagcaac gggcaaccgt ttggggaaag accacaccca cgcgcgatcg
     2221 ccatggcaac gaggtcgcac acgccccaca cccagacctc cctgcgagcg ggcatgggta
     2281 caatgtcccc gttgccacag agaccacttc gtagcacagc gcagagcgta gcgtgttgtt
     2341 gctgctgaca aaagaaaatt tttcttagca aagcaaagga ggggaagcac gggcagatag
     2401 caccgtacca tacccttgga aactcgaaat gaacgaagca ggaaatgaga gaatgagagt
     2461 tttgtaggta tatatagcgg tagtgtttgc gcgttaccat catcttctgg atctatctat
     2521 tgttcttttc ctcatcactt tccccttttt cgctcttctt cttgtctttt atttctttct
     2581 tttttttaat tgttccctcg attggctatc taccaaagaa tccaaactta atacacgtat
     2641 ttatttgtcc aattaccgga tcccccgggc tgcaggaatt catgtccaga ttagataaaa
     2701 gtaaagtgat taacagcgca ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa
     2761 cccgtaaact cgcccagaag ctaggtgtag agcagcctac attgtattgg catgtaaaaa
     2821 ataagcgggc tttgctcgac gccttagcca ttgagatgtt agataggcac catactcact
     2881 tttgcccttt agaaggggaa agctggcaag attttttacg taataacgct aaaagtttta
     2941 gatgtgcttt actaagtcat cgcgatggag caaaagtaca tttaggtaca cggcctacag
     3001 aaaaacagta tgaaactctc gaaaatcaat tagccttttt atgccaacaa ggtttttcac
     3061 tagagaatgc attatatgca ctcagcgctg tggggcattt tactttaggt tgcgtattgg
     3121 aagatcaaga gcatcaagtc gctaaagaag aaagggaaac acctactact gatagtatgc
     3181 cgccattatt acgacaagct atcgaattat ttgatcacca aggtgcagag ccagccttct
     3241 tattcggcct tgaattgatc atttgcggat tagaaaaaca acttaaatgt gaaagtgccc
     3301 ccaagaaaaa gagaaaggtg gcagaagccg ccgcaaagga agcggcagca aaagaggctg
     3361 cggccaagga agctgctgcc aaggcgctag aagcagaggc ggccgcaaag gaggcggccg
     3421 cgaaggaagc ggctgcgaag gaggcagcgg caaaggcgat gactgccagc gtttcgaata
     3481 cgcagaataa gctgaatgag cttctcgatg ccatcagaca ggagtttctc caagtctcac
     3541 aagaggcaaa tacctaccgt cttcaaaacc aaaaggatta cgatttcaaa atgaaccagc
     3601 agctggctga gatgcagcag ataagaaaca ccgtctacga actggaacta actcacagga
     3661 aaatgaagga cgcgtacgaa gaagagatca agcacttgaa actagggctg gagcaaagag
     3721 accatcaaat tgcatctttg accgtccagc aacagcggca acagcaacag cagcaacagg
     3781 tccagcagca tttacaacag caacagcagc agctagccgc tgcatctgca tctgttccag
     3841 ttgcgcaaca accaccggct actacttcgg ccaccgccac tccagcagca aacacaacta
     3901 ctggttcgcc atcggccttc ccagtacaag ctagccgtcc taatctggtt ggctcacagt
     3961 tgcctaccac cactttgcct gtggtgtcct caaacgccca acaacaacta ccacaacagc
     4021 aactgcaaca gcagcaactt caacaacagc aaccacctcc ccaggtttcc gtggcaccat
     4081 tgagtaacac agccatcaac ggatctccta cttctaaaga gaccactact ttaccctctg
     4141 tcaaggcacc tgaatctacg ttgaaagaaa ctgaaccgga aaataataat acctcgaaga
     4201 taaatgacac cggatccgcc accacggcca ccactaccac cgcaactgaa actgaaatca
     4261 aacctaagga ggaagacgcc accccggcta gtttgcacca ggatcactac ttagtccctt
     4321 ataatcaaag agcaaaccac tctaaaccta tcccaccttt ccttttggat ctagattccc
     4381 agtctgttcc cgatgctctg aagaagcaaa caaatgatta ttatatttta tacaacccgg
     4441 cactaccaag agaaattgac gttgagttac acaaatcttt ggatcatact tcagttgttt
     4501 gttgcgtgaa gttcagtaac gatggtgaat acttagccac aggctgcaac aaaactactc
     4561 aagtgtatcg cgtttcagat ggttctctgg tggcccgtct atctgacgat tctgctgcca
     4621 ataaccatcg aaattcgatc actgaaaata acaccaccac gtccacggat aacaatacaa
     4681 tgacaaccac tactaccacc acaattacta ccacagcgat gacttcggca gcagaattgg
     4741 caaaagatgt ggaaaacctg aacacttcgt cttccccatc atccgacttg tatatccgtt
     4801 cagtgtgttt ttctccagat gggaaatttt tggcaacagg tgctgaagac agactgatta
     4861 gaatttggga tattgaaaat agaaagattg ttatgattct tcaaggccac gaacaagata
     4921 tttattcatt ggactacttt ccctcaggtg acaaattagt ctccggttct ggtgaccgta
     4981 ccgttcgtat ttgggactta cgtacaggcc agtgttcatt gactttatcc attgaagatg
     5041 gtgttaccac cgtcgctgta tcaccaggtg atggtaaata catcgctgct ggttctctag
     5101 atcgtgctgt gagagtttgg gattccgaga ccggattctt ggtggaaaga ctagattcgg
     5161 aaaacgaatc cggtacaggc cacaaggact ctgtttatag cgttgtcttc actagagatg
     5221 gacaaagcgt tgtatccggc tcattagata gatctgttaa gctctggaat ttgcagaatg
     5281 caaacaacaa gagcgattcg aaaactccaa attccggcac ttgtgaagtt acgtatatcg
     5341 ggcataaaga ctttgtattg tccgtggcca ccacacaaaa tgatgagtac atcttgtccg
     5401 gttccaaaga tcgtggtgtc ctgttttggg ataagaaatc cggcaatccg ttattgatgt
     5461 tgcaaggtca taggaattca gttatatctg tggctgtggc aaacgggtct ccgctgggtc
     5521 cagaatataa cgtttttgct actggtagcg gtgattgtaa agcaaggatt tggaagtata
     5581 aaaaaatagc gccaaattaa ctcgagacag gccccttttc ctttgtctat atcatgtaat
     5641 tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag
     5701 gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt
     5761 aagaacgtta tttatatttc aaatttttct tttttttctg tacaaacgcg tgtacgcatg
     5821 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
     5881 caaggttcgc agtttacact ctcatcgtcg ctctcatcat cgcttccgtt gttgttttcc
     5941 ttagtagcgt ctgctttcta gacagttcga gtttatcatt atcaatactg ccatttcaaa
     6001 gaatacgtaa ataattaata gtagtgattt tcctaacttt atttagtcaa aaaattagcc
     6061 tctatcattg atagagtgtc tgggtgatct atcattgata gagcatccac taattttaat
     6121 tctgctgtaa cccgtacatg cccaaaatag ggggcgggtt acacagaatc tatcattgat
     6181 agagtgtctg ggtgatctat cattgataga gcatccacta aatataatgg agctctatca
     6241 ttgatagagc atccactaaa aaaaagaatc ccagcaccaa aatattgttt tcttcaccaa
     6301 ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc acaaacaggc
     6361 aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat gatgacacaa
     6421 ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta ccttctgctc
     6481 tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa attattcccc
     6541 tatctatcat tgatagatat aaatatctat cattgataga gtaattctgt aaatctattt
     6601 cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta aaacaccaag
     6661 aacttagttt cgaataaaca cacataaaca aacaaaatgt ccaggttgga caagtccaag
     6721 gttattaact ctgccttgga gttgttgaac gaagttggta ttgaaggttt gaccactaga
     6781 aagttggctc aaaaattggg tgttgaacaa cctaccttgt actggcatgt taagaacaaa
     6841 agagctttgt tggacgcttt ggccattgaa atgttggata gacatcatac ccatttctgt
     6901 ccattggaag gtgaatcatg gcaagatttc ttgagaaaca acgccaagtc tttcagatgt
     6961 gctttgttgt ctcatagaga tggtgctaaa gttcacttgg gtactagacc aactgaaaag
     7021 caatacgaaa ctttggaaaa ccagttggct ttcttgtgtc aacaaggttt ctctttagaa
     7081 aacgccttgt atgctttgtc tgctgttggt cattttacct tgggttgtgt tttggaagat
     7141 caagaacatc aagtcgccaa agaagaaagg gaaactccaa ctactgattc tatgccacca
     7201 ttattgagac aagccattga gttgtttgat catcaaggtg ctgaaccagc ttttttgttt
     7261 ggtttggaat tgatcatctg cggtttggag aaacaattga agtgtgaatc tgcccctaag
     7321 aagaagagaa aggtttgaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     7381 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     7441 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     7501 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     7561 ctcgaaggct ttaatttggg ccgtatttac agataacgat gtcgttatta tatatatata
     7621 tatatattgt cacttgtacc cagcttttgt tccctttagt gagggttaat ttcgagcttg
     7681 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aacagctgga
     7741 tgaattgaat agctttatgg accctgaaac cacagccaca ttaaccttct ttgatggtca
     7801 aaacttatcc ttcaccataa atatgcctcg caaaaaaggt aattaacata tatagaatta
     7861 cattatttat gaaatatcat cactatctct tagcatcttt aatccttttc tacatcagat
     7921 aacttcggtt tgttatcatc gtctgtattg tcatcaattg gcgcgcctct ccctgcaggc
     7981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8821 gtttaaacct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8881 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     8941 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     9001 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     9061 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     9121 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     9181 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     9241 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     9301 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     9361 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     9421 gatggttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     9481 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     9541 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     9601 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9661 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     9721 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     9781 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     9841 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     9901 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     9961 agtgccacct
//
