LOCUS       Exported                9970 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Repressor plasmid for WTC 846. Encodes TetR and TetR-Tup1, Ura3 
            selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    FRP2365
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9970)
  TITLE     The Well-Tempered Controller 846: Plasmids for tight and precise 
            control of gene expression in S.cerevisiae
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9970)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9970
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(954..973)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             1499..1519
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      3831..4080
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     protein_bind    4259..4276
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    4432..4448
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             5514..5534
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5577..5766
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(5852..5870)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5891..5907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5891..5907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5904..5926)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5915..5931
                     /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(6241..6258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6412..7000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6492..6511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7179..8039)
                     /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     7802..7821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8040..8144)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      8509..8706
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(9525..9868)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     9956..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9970..17
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gagaaacaga
       61 ataatatata aacctgtata atataacctt gaagactata tttcttttcg cggccgccta
      121 tagggcgaat tggagctcca ccgcggtggc tctagaagtc gaacaagaag caggcaaagt
      181 ttagagcact gcccctccgc actcaaaaaa gaaaaaacta ggaggaaaat aaaattctca
      241 accacacaaa cacataaaca catacaaata caaatacaag cttatttact tgacatcgcg
      301 cgatcttcca ctattcagcg ccgtccgccc tctctcgtgt tttttgttta cgcgacaact
      361 atgcgaaatc cggagcaacg ggcaaccgtt tggggaaaga ccacacccac gcgcgatcgc
      421 catggcaacg aggtcgcaca cgccccacac ccagacctcc ctgcgagcgg gcatgggtac
      481 aatgtccccg ttgccacaga gaccacttcg tagcacagcg cagagcgtag cgtgttgttg
      541 ctgctgacaa aagaaaattt ttcttagcaa agcaaaggag gggaagcacg ggcagatagc
      601 accgtaccat acccttggaa actcgaaatg aacgaagcag gaaatgagag aatgagagtt
      661 ttgtaggtat atatagcggt agtgtttgcg cgttaccatc atcttctgga tctatctatt
      721 gttcttttcc tcatcacttt cccctttttc gctcttcttc ttgtctttta tttctttctt
      781 ttttttaatt gttccctcga ttggctatct accaaagaat ccaaacttaa tacacgtatt
      841 tatttgtcca attaccggat cccccgggct gcaggaattc atgtccagat tagataaaag
      901 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac
      961 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa
     1021 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt
     1081 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag
     1141 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga
     1201 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact
     1261 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga
     1321 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc
     1381 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt
     1441 attcggcctt gaattgatca tttgcggatt agaaaaacaa cttaaatgtg aaagtgcccc
     1501 caagaaaaag agaaaggtgg cagaagccgc cgcaaaggaa gcggcagcaa aagaggctgc
     1561 ggccaaggaa gctgctgcca aggcgctaga agcagaggcg gccgcaaagg aggcggccgc
     1621 gaaggaagcg gctgcgaagg aggcagcggc aaaggcgatg actgccagcg tttcgaatac
     1681 gcagaataag ctgaatgagc ttctcgatgc catcagacag gagtttctcc aagtctcaca
     1741 agaggcaaat acctaccgtc ttcaaaacca aaaggattac gatttcaaaa tgaaccagca
     1801 gctggctgag atgcagcaga taagaaacac cgtctacgaa ctggaactaa ctcacaggaa
     1861 aatgaaggac gcgtacgaag aagagatcaa gcacttgaaa ctagggctgg agcaaagaga
     1921 ccatcaaatt gcatctttga ccgtccagca acagcggcaa cagcaacagc agcaacaggt
     1981 ccagcagcat ttacaacagc aacagcagca gctagccgct gcatctgcat ctgttccagt
     2041 tgcgcaacaa ccaccggcta ctacttcggc caccgccact ccagcagcaa acacaactac
     2101 tggttcgcca tcggccttcc cagtacaagc tagccgtcct aatctggttg gctcacagtt
     2161 gcctaccacc actttgcctg tggtgtcctc aaacgcccaa caacaactac cacaacagca
     2221 actgcaacag cagcaacttc aacaacagca accacctccc caggtttccg tggcaccatt
     2281 gagtaacaca gccatcaacg gatctcctac ttctaaagag accactactt taccctctgt
     2341 caaggcacct gaatctacgt tgaaagaaac tgaaccggaa aataataata cctcgaagat
     2401 aaatgacacc ggatccgcca ccacggccac cactaccacc gcaactgaaa ctgaaatcaa
     2461 acctaaggag gaagacgcca ccccggctag tttgcaccag gatcactact tagtccctta
     2521 taatcaaaga gcaaaccact ctaaacctat cccacctttc cttttggatc tagattccca
     2581 gtctgttccc gatgctctga agaagcaaac aaatgattat tatattttat acaacccggc
     2641 actaccaaga gaaattgacg ttgagttaca caaatctttg gatcatactt cagttgtttg
     2701 ttgcgtgaag ttcagtaacg atggtgaata cttagccaca ggctgcaaca aaactactca
     2761 agtgtatcgc gtttcagatg gttctctggt ggcccgtcta tctgacgatt ctgctgccaa
     2821 taaccatcga aattcgatca ctgaaaataa caccaccacg tccacggata acaatacaat
     2881 gacaaccact actaccacca caattactac cacagcgatg acttcggcag cagaattggc
     2941 aaaagatgtg gaaaacctga acacttcgtc ttccccatca tccgacttgt atatccgttc
     3001 agtgtgtttt tctccagatg ggaaattttt ggcaacaggt gctgaagaca gactgattag
     3061 aatttgggat attgaaaata gaaagattgt tatgattctt caaggccacg aacaagatat
     3121 ttattcattg gactactttc cctcaggtga caaattagtc tccggttctg gtgaccgtac
     3181 cgttcgtatt tgggacttac gtacaggcca gtgttcattg actttatcca ttgaagatgg
     3241 tgttaccacc gtcgctgtat caccaggtga tggtaaatac atcgctgctg gttctctaga
     3301 tcgtgctgtg agagtttggg attccgagac cggattcttg gtggaaagac tagattcgga
     3361 aaacgaatcc ggtacaggcc acaaggactc tgtttatagc gttgtcttca ctagagatgg
     3421 acaaagcgtt gtatccggct cattagatag atctgttaag ctctggaatt tgcagaatgc
     3481 aaacaacaag agcgattcga aaactccaaa ttccggcact tgtgaagtta cgtatatcgg
     3541 gcataaagac tttgtattgt ccgtggccac cacacaaaat gatgagtaca tcttgtccgg
     3601 ttccaaagat cgtggtgtcc tgttttggga taagaaatcc ggcaatccgt tattgatgtt
     3661 gcaaggtcat aggaattcag ttatatctgt ggctgtggca aacgggtctc cgctgggtcc
     3721 agaatataac gtttttgcta ctggtagcgg tgattgtaaa gcaaggattt ggaagtataa
     3781 aaaaatagcg ccaaattaac tcgagacagg ccccttttcc tttgtctata tcatgtaatt
     3841 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg
     3901 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta
     3961 agaacgttat ttatatttca aatttttctt ttttttctgt acaaacgcgt gtacgcatgt
     4021 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     4081 aaggttcgca gtttacactc tcatcgtcgc tctcatcatc gcttccgttg ttgttttcct
     4141 tagtagcgtc tgctttctag acagttcgag tttatcatta tcaatactgc catttcaaag
     4201 aatacgtaaa taattaatag tagtgatttt cctaacttta tttagtcaaa aaattagcct
     4261 ctatcattga tagagtgtct gggtgatcta tcattgatag agcatccact aattttaatt
     4321 ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta cacagaatct atcattgata
     4381 gagtgtctgg gtgatctatc attgatagag catccactaa atataatgga gctctatcat
     4441 tgatagagca tccactaaaa aaaagaatcc cagcaccaaa atattgtttt cttcaccaac
     4501 catcagttca taggtccatt ctcttagcgc aactacagag aacaggggca caaacaggca
     4561 aaaaacgggc acaacctcaa tggagtgatg caacctgcct ggagtaaatg atgacacaag
     4621 gcaattgacc cacgcatgta tctatctcat tttcttacac cttctattac cttctgctct
     4681 ctctgatttg gaaaaagctg aaaaaaaagg ttgaaaccag ttccctgaaa ttattcccct
     4741 atctatcatt gatagatata aatatctatc attgatagag taattctgta aatctatttc
     4801 ttaaacttct taaattctac ttttatagtt agtctttttt ttagttttaa aacaccaaga
     4861 acttagtttc gaataaacac acataaacaa acaaaatgtc caggttggac aagtccaagg
     4921 ttattaactc tgccttggag ttgttgaacg aagttggtat tgaaggtttg accactagaa
     4981 agttggctca aaaattgggt gttgaacaac ctaccttgta ctggcatgtt aagaacaaaa
     5041 gagctttgtt ggacgctttg gccattgaaa tgttggatag acatcatacc catttctgtc
     5101 cattggaagg tgaatcatgg caagatttct tgagaaacaa cgccaagtct ttcagatgtg
     5161 ctttgttgtc tcatagagat ggtgctaaag ttcacttggg tactagacca actgaaaagc
     5221 aatacgaaac tttggaaaac cagttggctt tcttgtgtca acaaggtttc tctttagaaa
     5281 acgccttgta tgctttgtct gctgttggtc attttacctt gggttgtgtt ttggaagatc
     5341 aagaacatca agtcgccaaa gaagaaaggg aaactccaac tactgattct atgccaccat
     5401 tattgagaca agccattgag ttgtttgatc atcaaggtgc tgaaccagct tttttgtttg
     5461 gtttggaatt gatcatctgc ggtttggaga aacaattgaa gtgtgaatct gcccctaaga
     5521 agaagagaaa ggtttgaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc
     5581 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     5641 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     5701 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     5761 tcgaaggctt taatttgggc cgtatttaca gataacgatg tcgttattat atatatatat
     5821 atatattgtc acttgtaccc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     5881 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca acagctggat
     5941 gaattgaata gctttatgga ccctgaaacc acagccacat taaccttctt tgatggtcaa
     6001 aacttatcct tcaccataaa tatgcctcgc aaaaaaggta attaacatat atagaattac
     6061 attatttatg aaatatcatc actatctctt agcatcttta atccttttct acatcagata
     6121 acttcggttt gttatcatcg tctgtattgt catcaattgg cgcgcctctc cctgcaggca
     6181 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6241 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6301 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6361 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6421 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6481 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6541 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6601 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6661 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6721 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6781 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6841 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6901 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6961 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttg
     7021 tttaaaccta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7081 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7141 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7201 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7261 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7321 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7381 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7441 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7501 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7561 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7621 atggttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7681 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7741 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7801 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7861 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7921 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7981 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8041 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8101 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8161 gtgccacctg acgtcgaaag aacgcggcat tagcaacatt tgcgaagcga acgattttgc
     8221 gaactgctat taattaactc tggcgcgcca ctactttttg gaactcttgt tgttctttgg
     8281 agttcaatgc gtccatcttt acagtcctgt cttattgttc ttgatttgtg ccccgtaaaa
     8341 tactgttact tggttctggc gaggtattgg atagttcctt tttataaagg ccatgaaggt
     8401 ttttctttcc aatttttttt ttttcgtcat tatagaaatc attacgaccg agattcccgg
     8461 catatgcgtt ttcgacactg gatggcggcg ttagtatcga atcgacagca gtatagcgac
     8521 cagcattcac atacgattga cgcatgatat tactttctgc gcacttaact tcgcatctgg
     8581 gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta acatttgatt aaaatagaac
     8641 aactacaata taaaaaaact atacaaatga caagttcttg aaaacaagaa tctttttatt
     8701 gtcagtactg attataattg gccagtcttt ttcaaataag cattccaacc agcatctcta
     8761 taccttttac cttcaatatc tggatctctt cctttaccaa acaatcctct accaacaatg
     8821 ataatatcag ttccagtgct aacaacttca tcaacagttc tatattgttg tcctaatcca
     8881 tcacctttat catctaatcc aactccaggt gtcataataa gccaatcaaa tccttcttct
     8941 tgtccaccca tatcacgttg ggcaataaat ccaataacaa attccttatc ggatttagca
     9001 atttcaacag ttttttgaga atattctcca taagctaatg atcccactga tgataattca
     9061 gctaacatca ataaccctct tggctcttgg ttggtggtgg tttctttagc tccctgtttt
     9121 aatccttcaa ctactccatt cccagtgaca ccatgagcat tagtaatatc tgcccaacta
     9181 ctaattttat aaactccacc aatatattgt ttcttcacgg tattaccaat atcagcaaat
     9241 tttctatctt caaaaatcat aaattgatgt ttacgtgaaa gttctaataa tggttcaata
     9301 gtggattcat aggaaaaatc attgattata tcaatatgag tcttgattaa gcatacataa
     9361 ggacccaatt tatcaattaa ttcaagaaat tccttagtgg tatcaacatc aattgatgca
     9421 cataaattgg ttttcttcag ttccattaat cgaaataatc gttgtgctac tggtgaggca
     9481 tgagtttctg ctctctcact ataggtctta gtgttgactg tcatggttgt ttatgttcgg
     9541 atgtgatgtg agaactgtat cctagcaaga ttttaaaagg aagtatatga aagaagaacc
     9601 tcagtggcaa atcctaacct tttatatttc tctacagggg cgcggcgtgg ggacaattca
     9661 acgcgtctgt gaggggagcg tttccctgct cgcaggtccg cagcgaggag ccgtaatttt
     9721 tgcttcgcgc cgtgcggcca tcaaaatgta tggatgcaaa tgattataca tggggatgta
     9781 tgggctaaat gtacgggcga cagtcacatc atgcccctga gctgcgcacg tcaagactgt
     9841 caaggagggt attctgggcc tccatgtcgc tggccgggtg acccggcggg gacgaggcaa
     9901 gctaaaccga tcggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9961 tcacgacgtt
//
