LOCUS       sequence-370759-        8969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing artificial trans-activator in Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8969)
  AUTHORS   Peng B, Bandari NC, Lu Z, Howard CB, Scott C, Trau M, Dumsday G, 
            Vickers CE
  TITLE     Engineering eukaryote-like regulatory circuits to expand artificial 
            control mechanisms for metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Commun Biol. 2022 Feb 16;5(1):135. doi: 10.1038/s42003-022-03070-z.
  PUBMED    35173283
REFERENCE   2  (bases 1 to 8969)
  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: "Commun 
            Biol."; date: "2022-02-16"; volume: "5(1):135. doi"; pages: " 
            10.1038/s42003-022-03070-z"
COMMENT     Sequence Label: pAFZifPZ432
FEATURES             Location/Qualifiers
     source          1..8969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             350..370
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3814..3833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3815..3833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3843..3860)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3843..3859)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3852..3874)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4000..4455
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4087..4106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4297..4318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4558..5232)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(5233..5513)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5534..5553)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5653..5675
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5713..5731)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5772..6275
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6312..6416
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6417..6485,6486..7277)
                     /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(6635..6654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7448..8036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7937..7956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8190..8207
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8324..8345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8360..8365,8366..8383,8384..8390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8398..8414
                     /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     8403..8425
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8422..8438
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8422..8438
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8457..8477
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8459..8477
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cccacctacg acaacaaccg ccactccgcg gatcatgggt actagaccat acgcttttaa
       61 gtgtccagtt tgtggtaaag ccttcagaca ttcttcatcc ttggttagac atcaaagaac
      121 tcatactggt caaaagccat acaagtgtaa gcaatgtgga aaggcttttg gttgtccatc
      181 taatttgaga aggcatggta gaacccatac aggtgaaaaa ccttatgaat gtcacgattg
      241 tggcaagtct ttcagacaat ctactcattt gactcagcat agaagaatcc acaccggtgg
      301 tggtggtact ccagctgctg cttctacttt ggaatctaga tgtaccccac ctaagaagaa
      361 aagaaaggtt gctagctctg ctgctcctgt ccaagacaaa gacactctgt ccaatgccga
      421 gcgtgcgaag aacgtcaacg ggttgcttca ggtgctcatg gacattaaca ctctgaacgg
      481 agggagctcc gacactgctg ataagataag aattcatgcc aaaaacttcg aggcagcttt
      541 gttcgcaaag agctcttcaa agaaagaata catggacagc atgaacgaaa aagttgctgt
      601 catgcgcaac acgtacaata cgaggaaaaa cgccgttact gctgctgccg ctaataacaa
      661 cattaaaccc gtggaacagc accatattaa caacttgaaa aattctggca acagcgccaa
      721 taatatgaat gtcaatatga atctaaaccc acagatgttc ttgaatcagc aggctcaggc
      781 aaggcaacag gttgcgcaac aattaagaaa tcaacaacaa caacaacaac agcagcagca
      841 gcaacagagg cgtcaattga ctcctcaaca acaacaatta gtgaaccaga tgaaagtggc
      901 acctattccc aaacaattac tgcaaagaat tcctaacatt ccacccaata tcaacacctg
      961 gcagcaggtc actgctttgg ctcaacaaaa gctattgaca cctcaggata tggaagctgc
     1021 gaaggaagtc tacaagattc accagcagtt gctattcaaa gcaaggctac agcaacaaca
     1081 agcacaggct caagcacaag ctaataacaa caacaacggc ctcccccaaa atggtaatat
     1141 taacaataac ataaatattc ctcaacagca gcaaatgcaa cctcccaatt caagtgcgaa
     1201 caacaaccct ttgcaacagc aatcatcaca aaataccgta ccaaacgtcc tcaaccaaat
     1261 taaccaaatc ttttctccag aggagcaacg cagcttatta caagaagcca tcgaaacctg
     1321 caagaatttt gaaaaaacac aattgggtag tacgatgacg gaacctgtca agcaaagttt
     1381 tattaggaaa tacattaacc aaaaggccct gagaaaaatc caagctttga gagatgttaa
     1441 gaacaacaat aacgctaaca acaacggctc gaaccttcag agagcccaaa atgtccctat
     1501 gaatatcatc caacaacaac aacaacagaa cacgaacaat aatgacacca ttgccacttc
     1561 tgctacacct aatgctgccg ctttctctca gcaacagaac gcaagttcta aattatatca
     1621 gatgcaacaa cagcaacaag ctcaagctca agctcaagct caagctcagg cacaggctca
     1681 agcacaagct caagcacagg cggcacaagc ggcgcaagcg caagcacaag cacaagcaca
     1741 agcacaagca caggcacagg cacaggcaca agcccaggcg caggcccaag cccaagccca
     1801 agcacaagca caagcacacg ctcagcacca gccctcccaa caaccacaac aagctcagca
     1861 acaacctaac ccactacatg ggttgacacc tactgcaaag gatgtcgaag taattaagca
     1921 attgtccttg gatgcttcta agaccaacct aaggcttacg gacgtaacaa attctttatc
     1981 caatgaagaa aaggaaaaaa ttaaaatgaa gttaaagcaa ggtcaaaagc tttttgttca
     2041 ggtgagtaat ttcgccccac aagtctacat catcacaaag aatgaaaact tcttgaagga
     2101 agtttttcag ttaagaatct ttgtaaaaga gatcctagaa aaatgtgccg agggtatatt
     2161 tgttgttaaa ttagacaccg ttgacaggtt aattattaag tatcaaaaat attgggaaag
     2221 tatgagaatt caaattttaa gaagacaagc tattttaaga caacaacagc aaatggcaaa
     2281 caacaatggg aacccaggca ctacttctac tggaaacaat aataatattg caactcagca
     2341 aaatatgcaa cagtcactac agcaaatgca gcatttacag caattgaaaa tgcagcagca
     2401 acaacaacag cagcaacaac aacagcagca acaacaacag cagcaacaac aacagcaaca
     2461 gcacatatat ccctcctcga ctcctggtgt ggctaattat tcggcaatgg ctaatgcacc
     2521 cggtaacaat atcccatata tgaaccataa aaatacctct agcatggatt ttttgaactc
     2581 tatggaaaat acaccaaaag ttcccgtatc cgctgcggca accccatcac tgaacaagac
     2641 gatcaacggt aaggtgaatg gcaggacaaa atctaatacg atacctgtta ccagcattcc
     2701 atcaacaaat aagaaactat caatttcaaa tgccgctagt caacaaccca ctcctaggtc
     2761 tgcatcaaat accgctaagt caaccccaaa tacaaatcct tctccactga aaacccaaac
     2821 taaaaatggc acaccaaacc ccaataatat gaagactgta cagtctccta tgggtgcaca
     2881 accatcatat aatagtgcca ttatagagaa tgcattcaga aaggaagaac tcttgttaaa
     2941 agatttggaa atcaggaagt tggagatatc ttctcgtttt aaacatcgcc aagaaatttt
     3001 caaagattct cctatggatt tgtttatgag tacgctgggt gattgcttag gtatcaaaga
     3061 tgaagagatg cttacgtcat gcactatccc aaaggctgtg gttgatcaca tcaacggctc
     3121 tggcaaaagg aagcctacaa aagcggctca aagggctcgc gatcaagatt ccattgacat
     3181 ttccataaaa gacaacaaat tggttatgaa gagtaaattc aataagagca ataggtcgta
     3241 ttcgatagcg ttgtccaatg tagctgctat attcaagggt atcggtggta actttaaaga
     3301 cttgtccact ttggttcatt catcatcgcc gtccacatct tctaatatgg atgtcggcaa
     3361 cccaagaaaa agaaaagcca gcgtattaga aataagcccg caagattcga tagcctcggt
     3421 gctatcacca gattcaaata taatgagtga ttctaaaaaa attaaagtag attctcctga
     3481 tgacccattc atgacaaaat caggagccac aactagtgaa aaacaagaag taacaaatga
     3541 agctccattt ttaacttctg ggactagttc agaacaattc aatgtatggg attggaataa
     3601 ttggacaagt gctacttaac ctgcaggata cccagttgag aagacgagct atttttctca
     3661 tgtttctttt gtttattcac ttttctatac tttttctttt ctattgaaat atatattata
     3721 tcttatgtat gtttttttgt attataaaca agcaaatttc gtcttgccat ttaccttctg
     3781 atattaatca catgtatatc gggccccaat tcgccctata gtgagtcgta ttacgcgcgc
     3841 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3901 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3961 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     4021 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4081 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4141 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4201 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4261 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4321 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4381 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4441 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4501 ccgcatagat cggcaagtgc acaaacaata cttaaataaa tactactcag taataaccta
     4561 tttcttagca tttttgacga aatttgctat tttgttagag tcttttacac catttgtctc
     4621 cacacctccg cttacatcaa caccaataac gccatttaat ctaagcgcat caccaacatt
     4681 ttctggcgtc agtccaccag ctaacataaa atgtaagctt tcggggctct cttgccttcc
     4741 aacccagtca gaaatcgagt tccaatccaa aagttcacct gtcccacctg cttctgaatc
     4801 aaacaaggga ataaacgaat gaggtttctg tgaagctgca ctgagtagta tgttgcagtc
     4861 ttttggaaat acgagtcttt taataactgg caaaccgagg aactcttggt attcttgcca
     4921 cgactcatct ccatgcagtt ggacgatatc aatgccgtaa tcattgacca gagccaaaac
     4981 atcctcctta ggttgattac gaaacacgcc aaccaagtat ttcggagtgc ctgaactatt
     5041 tttatatgct tttacaagac ttgaaatttt ccttgcaata accgggtcaa ttgttctctt
     5101 tctattgggc acacatataa tacccagcaa gtcagcatcg gaatccaaag cacattctgc
     5161 ggcctctgtg ctctgcaagc cgcaaacttt caccaatgga ccagaactac ctgtgaaatt
     5221 aataacagac atactccaag ctgcctttgt gtgcttaatc acgtatactc acgtgctcaa
     5281 tagtcaccaa tgccctccct cttggccctc tccttttctt ttttcgaccg aattaattct
     5341 taatcggcaa aaaaagaaaa gctccggatc aagattgtac gtaaggtgac aagctatttt
     5401 tcaataaaga atatcttcca ctactgccat ctggcgtcat aactgcaaag tacacatata
     5461 ttacgatgct gttctattaa atgcttccta tattatatat atagtaatgt cgtgatctat
     5521 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     5581 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5641 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5701 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5761 tttcttagac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa
     5821 taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg attttagaaa
     5881 gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa
     5941 aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa
     6001 tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt
     6061 cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga
     6121 taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg aaaacaaaaa
     6181 ctattttttc tttaatttct ttttttactt tctattttta atttatatat ttatattaaa
     6241 aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt ggcacttttc
     6301 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6361 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6421 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6481 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6541 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6601 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6661 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6721 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6781 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6841 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6901 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6961 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7021 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7081 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7141 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7201 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7261 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7321 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7381 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7441 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7501 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7561 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7621 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7681 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7741 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7801 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7861 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7921 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7981 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8041 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8101 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8161 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8221 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     8281 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     8341 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     8401 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     8461 ttaaccctca ctaaagggaa caaaagctgg agctctacaa tgaaaacaac accaagctgc
     8521 caaaagctgt atattaaagt taaagctccc tcatttttag taaggacgct ttcttttccc
     8581 acaaaaggga cgaggaaaaa aagaaaaacc tggaaatata gtcacgtgac atgaataaac
     8641 aagacctagg gcaatatgga aaaagaagct aggataaaaa tacatttatg agggttgtaa
     8701 ggcaaagtgg ctcagcatta agtagggcta aaaagagtcg caaatataca tcgtaccgag
     8761 tgatgaacgt ggttttgaac accttgttct cttttgttct cgctccctac attcactata
     8821 tattagaaga aatttcacct caatggacaa ctcgagaaat gaatacagaa atatgttttt
     8881 tagcgaaatt ttcctttctt cttgtcttct tgttttattt aaacttccaa ggctttaact
     8941 cagtgtcaaa cataacaacc tcctcctcc
//
