LOCUS       sequence-370741-        8903 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 8903)
  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 8903)
  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: pAF9D91
FEATURES             Location/Qualifiers
     source          1..8903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             66..86
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3101..3120)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3102..3120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3130..3147)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3130..3146)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3139..3161)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3287..3742
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3374..3393)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3584..3605
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3845..4519)
                     /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(4520..4800)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4821..4840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4940..4962
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5000..5018)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5059..5562
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5599..5703
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5704..5772,5773..6564)
                     /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(5922..5941)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6735..7323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7224..7243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7477..7494
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7611..7632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7647..7652,7653..7670,7671..7677)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7685..7701
                     /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     7690..7712
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7709..7725
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7709..7725
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7744..7764
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7746..7764
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctggctccgg taccggtggt ggtactccag ctgctgcttc tactttggaa tctagatgta
       61 ccccacctaa gaagaaaaga aaggttgcta gcagaacgtc tctcactgaa cagtacgctc
      121 cttctagtat acaacatcag cagcagcacc cgcaaagtat tagtcctata gcaaacacgc
      181 agtatggtga tatccctgtt agaccggaaa ttgatttaga tcctagcatt gtgcctgtgg
      241 tccctgaacc cactgagccc atcgaaaaca atatatcgct taatgaggaa gtcactttct
      301 tcgaaaaggc caagaggtat atcggcaata aacatttata cactgagttt ttgaaaattt
      361 taaatttgta ctctcaagat atacttgatc ttgacgattt agtggaaaag gtagatttct
      421 acttgggttc caataaagaa ctatttacgt ggttcaaaaa ctttgttggc taccaagaaa
      481 aaaccaaatg tatcgagaat attgttcatg aaaaacatag attggattta gatttatgtg
      541 aggcatttgg cccaagttac aagaggctac caaaaagtga cactttcatg ccatgctcag
      601 gtagggatga tatgtgttgg gaagtcttga acgatgaatg ggttggacat cctgtatggg
      661 cttccgaaga ttcgggattt attgctcatc gtaaaaacca gtatgaggaa acactattca
      721 agatcgaaga ggaaagacat gagtatgatt tttacattga atcaaattta agaactattc
      781 aatgtttgga aacaattgta aataagatcg agaacatgac tgagaatgaa aaagccaatt
      841 ttaaactgcc tccaggtctt ggtcatactt caatgactat ttataaaaaa gtgataagga
      901 aagtttatga taaggaaagg gggttcgaga ttattgatgc tttgcatgag caccctgcag
      961 tgacagcccc agtcgttctg aaaagattaa agcaaaaaga cgaagaatgg agaagagctc
     1021 aacgtgaatg gaataaagtt tggagggagt tagaacagaa ggtttttttc aagtcattag
     1081 atcatttagg cttaacattt aaacaggctg acaagaaatt attaactaca aagcagttga
     1141 tatcagagat tagcagcatc aaagttgatc aaacaaacaa aaaaattcac tggttaactc
     1201 ctaaaccaaa gagccagtta gattttgatt tccctgataa aaacattttc tatgatatct
     1261 tgtgtttggc tgacactttt ataacccata ccacagccta ttctaatccc gataaagaaa
     1321 gattgaaaga tttactaaaa tacttcatct ctttgttttt ttctatttct ttggaaaaaa
     1381 tcgaagaatc gttgtactcc cataagcaaa acgtgtcaga atctagcgga tctgacgatg
     1441 gcagttctat tgcatcaagg aagaggccct atcaacaaga aatgagttta cttgatattt
     1501 tacacaggag cagatatcaa aagctaaagc ggtctaatga tgaagatggc aaagttcccc
     1561 agctctctga accacccgaa gaagaaccta ataccattga ggaggaagag ctcatcgatg
     1621 aagaagctaa aaatccgtgg ctaactggga atttagtgga ggaagcaaac tctcagggta
     1681 ttatccaaaa tcgcagtatt tttaatctat tcgctaatac gaatatttac atatttttcc
     1741 gtcattggac aacgatttat gagcggcttt tggaaattaa gcaaatgaat gaaaaggtca
     1801 caaaggaaat caacacaagg tcgacagtta cttttgccaa agatctagat ttattatcga
     1861 gtcaactttc cgaaatggga ctagattttg ttggtgaaga cgcgtacaaa caggttttaa
     1921 gactgagtag aaggttgatt aatgggggtc ttgaacatca gtggtttgaa gagagtttgc
     1981 ggcaagctta caataacaaa gcgtttaaac tctatacaat tgataaagtc acccaatcgt
     2041 tggtaaagca tgctcatacc ttgatgactg acgctaaaac tgcagaaata atggctttgt
     2101 tcgttaaaga tagaaatgcc tccaccacga gtgcgaagga ccaaattatc tatcgcttgc
     2161 aggtgcgctc acatatgtcc aatacagaaa atatgtttag aatagagttt gataaaagaa
     2221 ctctgcatgt ttccattcaa tatattgcac ttgatgattt gacactaaag gaaccaaagg
     2281 cagacgaaga taaatggaaa tattatgtaa catcgtatgc tctgccacat ccaacagaag
     2341 gcattccgca cgagaaactg aagataccat ttttggaaag gctcatcgaa tttggacaag
     2401 atattgatgg aacagaggta gatgaggagt tttcccccga aggcatctct gtatcgacat
     2461 tgaaaattaa gatccagcct ataacctacc aactgcatat tgaaaacggg tcttacgatg
     2521 tttttacccg taaggctact aacaaatatc ctactattgc taatgataat actcaaaaag
     2581 gaatggtttc tcagaagaag gagttgatat caaaatttct agactgtgca gttggtttga
     2641 gaaataattt agatgaggct caaaaattaa gtatgcaaaa aaaatgggag aacctaaaag
     2701 acagcatagt aaagacgagt gctggcaacc agggaataga gagtgaaacc gaaaaaggta
     2761 aaattacgaa acaggagcag tcggataatt tggacagctc tactgcaagt gtactgcctg
     2821 catccattac tactgtacca caagatgata atatagaaac gactgggaat actgaatctt
     2881 cagacaaggg ggctaagatt caataacctg caggataccc agttgagaag acgagctatt
     2941 tttctcatgt ttcttttgtt tattcacttt tctatacttt ttcttttcta ttgaaatata
     3001 tattatatct tatgtatgtt tttttgtatt ataaacaagc aaatttcgtc ttgccattta
     3061 ccttctgata ttaatcacat gtatatcggg ccccaattcg ccctatagtg agtcgtatta
     3121 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3181 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3241 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     3301 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3361 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3421 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3481 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3541 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3601 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3661 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3721 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3781 tttcacaccg catagatcgg caagtgcaca aacaatactt aaataaatac tactcagtaa
     3841 taacctattt cttagcattt ttgacgaaat ttgctatttt gttagagtct tttacaccat
     3901 ttgtctccac acctccgctt acatcaacac caataacgcc atttaatcta agcgcatcac
     3961 caacattttc tggcgtcagt ccaccagcta acataaaatg taagctttcg gggctctctt
     4021 gccttccaac ccagtcagaa atcgagttcc aatccaaaag ttcacctgtc ccacctgctt
     4081 ctgaatcaaa caagggaata aacgaatgag gtttctgtga agctgcactg agtagtatgt
     4141 tgcagtcttt tggaaatacg agtcttttaa taactggcaa accgaggaac tcttggtatt
     4201 cttgccacga ctcatctcca tgcagttgga cgatatcaat gccgtaatca ttgaccagag
     4261 ccaaaacatc ctccttaggt tgattacgaa acacgccaac caagtatttc ggagtgcctg
     4321 aactattttt atatgctttt acaagacttg aaattttcct tgcaataacc gggtcaattg
     4381 ttctctttct attgggcaca catataatac ccagcaagtc agcatcggaa tccaaagcac
     4441 attctgcggc ctctgtgctc tgcaagccgc aaactttcac caatggacca gaactacctg
     4501 tgaaattaat aacagacata ctccaagctg cctttgtgtg cttaatcacg tatactcacg
     4561 tgctcaatag tcaccaatgc cctccctctt ggccctctcc ttttcttttt tcgaccgaat
     4621 taattcttaa tcggcaaaaa aagaaaagct ccggatcaag attgtacgta aggtgacaag
     4681 ctatttttca ataaagaata tcttccacta ctgccatctg gcgtcataac tgcaaagtac
     4741 acatatatta cgatgctgtt ctattaaatg cttcctatat tatatatata gtaatgtcgt
     4801 gatctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4861 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4921 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4981 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5041 ataatggttt cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat
     5101 gatggaataa tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt
     5161 ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag
     5221 gtttaaaaaa tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca
     5281 gattaaatag atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt
     5341 gtgtggtctt ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag
     5401 agcaagataa aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa
     5461 acaaaaacta ttttttcttt aatttctttt tttactttct atttttaatt tatatattta
     5521 tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc
     5581 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5641 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5701 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5761 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5821 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5881 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5941 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6001 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6061 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6121 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6181 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6241 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6301 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6361 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6421 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6481 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6541 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6601 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6661 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6721 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6781 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6841 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6901 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6961 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7021 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7081 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7141 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7201 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7261 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7321 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7381 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7441 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7501 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7561 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7621 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7681 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7741 cgcgcaatta accctcacta aagggaacaa aagctggagc tctacaatga aaacaacacc
     7801 aagctgccaa aagctgtata ttaaagttaa agctccctca tttttagtaa ggacgctttc
     7861 ttttcccaca aaagggacga ggaaaaaaag aaaaacctgg aaatatagtc acgtgacatg
     7921 aataaacaag acctagggca atatggaaaa agaagctagg ataaaaatac atttatgagg
     7981 gttgtaaggc aaagtggctc agcattaagt agggctaaaa agagtcgcaa atatacatcg
     8041 taccgagtga tgaacgtggt tttgaacacc ttgttctctt ttgttctcgc tccctacatt
     8101 cactatatat tagaagaaat ttcacctcaa tggacaactc gagaaatgaa tacagaaata
     8161 tgttttttag cgaaattttc ctttcttctt gtcttcttgt tttatttaaa cttccaaggc
     8221 tttaactcag tgtcaaacat aacaacctcc tcctccccca cctacgacaa caaccgccac
     8281 tccgcggatc atgtccagat tggataagtc caaggttatt aactctgcct tggaattgtt
     8341 gaacgaagtt ggtattgaag gtttgaccac tagaaagttg gctcaaaaat tgggtgttga
     8401 acaacctacc ttgtactggc atgttaagaa caaaagagct ttgttggacg ctttggccat
     8461 tgaaatgttg gatagacatc atacccattt ctgtccattg gaaggtgaat catggcaaga
     8521 tttcttgaga aacaacgcca agtctttcag atgtgctttg ttgtctcata gagatggtgc
     8581 taaagttcac ttgggtacta gaccaactga aaagcaatac gaaactttgg aaaaccagtt
     8641 ggctttcttg tgtcaacaag gtttctcttt agaaaacgcc ttgtatgctt tgtctgctgt
     8701 tggtcatttt accttgggtt gtgttttgga agatcaagaa catcaagtcg ccaaagaaga
     8761 aagggaaact ccaactactg attctatgcc accattattg agacaagcca ttgagttgtt
     8821 tgatcatcaa ggtgctgaac cagctttttt gtttggtttg gaattgatca tctgcggttt
     8881 ggagaaacag ttgaaatgtg aat
//
