LOCUS       sequence-319617-        8492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Aga1p in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8492)
  AUTHORS   Potts KA, Stieglitz JT, Lei M, Van Deventer JA
  TITLE     Reporter system architecture affects measurements of noncanonical 
            amino acid incorporation efficiency and fidelity.
  JOURNAL   Mol Syst Des Eng. 2020 Feb 1;5(2):573-588. doi: 10.1039/c9me00107g. 
            Epub 2020 Jan 23.
  PUBMED    33791108
REFERENCE   2  (bases 1 to 8492)
  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"; doi: 
            "10.1039/c9me00107g"; journalName: "Mol Syst Des Eng"; date: 
            "2020-02-1- 1"; volume: "5"; issue: "2"; pages: "573-588"
COMMENT     Sequence Label: pCTCON2-Aga1p
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             768..779
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             780..806
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1059..1088
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1421..1440)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1422..1440)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1447..1464)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1447..1463)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1456..1478)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1604..2059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1691..1710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1901..1922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2162..2836)
                     /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(2837..3117)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3138..3157)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3257..3279
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3317..3335)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3376..3879
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3916..4020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4021..4089,4090..4881)
                     /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(4239..4258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5052..5640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5541..5560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5794..5811
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5928..5949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5964..5969,5970..5987,5988..5994)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6002..6018
                     /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     6007..6029
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6026..6042
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6026..6042
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6063..6081
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(8289..8492,1..449)
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
ORIGIN
        1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
      361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaacc ccggatcgaa ttccctactt catacatttt
      481 caattaagat gcagttactt cgctgttttt caatattttc tgttattgct tcagttttag
      541 cacaggaact gacaactata tgcgagcaaa tcccctcacc aactttagaa tcgacgccgt
      601 actctttgtc aacgactact attttggcca acgggaaggc aatgcaagga gtttttgaat
      661 attacaaatc agtaacgttt gtcagtaatt gcggttctca cccctcaaca actagcaaag
      721 gcagccccat aaacacacag tatgttttta aggacaatag ctcgacgatt gaaggtagat
      781 acccatacga cgttccagac tacgctctgc aggctagtgg tggaggaggc tctggtggag
      841 gcggtagcgg aggcggaggg tcggctagct gcggtggcgg cggtactagc aaaatttctc
      901 attttttgaa aatggaatct ttgaatttta ttagagctca tactccatat attaatattt
      961 acaattgtga accagctaat ccatctgaaa aaaattctcc atctactcaa tattgttatt
     1021 ctattcaatc ttcccaggtc gactgcgggg gcggatccga acaaaagctt atttctgaag
     1081 aggacttgta atagctcgag atctgataac aacagtgtag atgtaacaaa atcgactttg
     1141 ttcccactgt acttttagct cgtacaaaat acaatatact tttcatttct ccgtaaacaa
     1201 catgttttcc catgtaatat ccttttctat ttttcgttcc gttaccaact ttacacatac
     1261 tttatatagc tattcacttc tatacactaa aaaactaaga caattttaat tttgctgcct
     1321 gccatatttc aatttgttat aaattcctat aatttatcct attagtagct aaaaaaagat
     1381 gaatgtgaat cgaatcctaa gagaattgag ctccaattcg ccctatagtg agtcgtatta
     1441 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1501 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     1561 cgatcgcctt tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     1621 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1681 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1741 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1801 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1861 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1921 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1981 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2041 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2101 cacaccgcat agatcggcaa gtgcacaaac aatacttaaa taaatactac tcagtaataa
     2161 cctatttctt agcatttttg acgaaatttg ctattttgtt agagtctttt acaccatttg
     2221 tctccacacc tccgcttaca tcaacaccaa taacgccatt taatctaagc gcatcaccaa
     2281 cattttctgg cgtcagtcca ccagctaaca taaaatgtaa gctttcgggg ctctcttgcc
     2341 ttccaaccca gtcagaaatc gagttccaat ccaaaagttc acctgtccca cctgcttctg
     2401 aatcaaacaa gggaataaac gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc
     2461 agtcttttgg aaatacgagt cttttaataa ctggcaaacc gaggaactct tggtattctt
     2521 gccacgactc atctccatgc agttggacga tatcaatgcc gtaatcattg accagagcca
     2581 aaacatcctc cttaggttga ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac
     2641 tatttttata tgcttttaca agacttgaaa ttttccttgc aataaccggg tcaattgttc
     2701 tctttctatt gggcacacat ataataccca gcaagtcagc atcggaatct agagcacatt
     2761 ctgcggcctc tgtgctctgc aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga
     2821 aattaataac agacatactc caagctgcct ttgtgtgctt aatcacgtat actcacgtgc
     2881 tcaatagtca ccaatgccct ccctcttggc cctctccttt tcttttttcg accgaattaa
     2941 ttcttaatcg gcaaaaaaag aaaagctccg gatcaagatt gtacgtaagg tgacaagcta
     3001 tttttcaata aagaatatct tccactactg ccatctggcg tcataactgc aaagtacaca
     3061 tatattacga tgctgttcta ttaaatgctt cctatattat atatatagta atgtcgtgat
     3121 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3181 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3241 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3301 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3361 atggtttctt agacggatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat
     3421 ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta
     3481 gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt
     3541 taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat
     3601 taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg
     3661 tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc
     3721 aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca
     3781 aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat atatttatat
     3841 taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc aggtggcact
     3901 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3961 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4021 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4081 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4141 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4201 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4261 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4321 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4381 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4441 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4501 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4561 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4621 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4681 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4741 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4801 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4861 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4921 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4981 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5161 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5401 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5641 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5701 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5761 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5821 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5881 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     5941 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6001 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     6061 gaaattaacc ctcactaaag ggaacaaaag ctgggtacct taactgaaaa ttacattgca
     6121 agcaactgcc atgatggcga agaccaactt tcctaatggc aaacgcgttt gcgatccact
     6181 gcccatatat tggctaattg ttaccattga tgaagtttgg tattgtatgg aggttgaagt
     6241 agtggaaact tctgtagtgg taacagatgt ttgtagagaa gagggagcag atgatgatga
     6301 gaggaaaata gaagataccg tcggagtcaa tgatgaagat tcaatagtcg attccacaga
     6361 ggtagatggt gtagagatag atgttgcgga aagtgatgat gtagaggcag atattgtagc
     6421 aatagctgtg gaagaagtgg gttgtaaaat ctctgatgtt gaagaaattt gaagtgtggt
     6481 aactggtgac tgttcgacac ttagtgatga tacagaactg aatagcaaaa tagtactgtc
     6541 agaggataat gtagcagaca aggaggagga tgaaaggacc gatgtcgata ataagtgtga
     6601 tgattgttct tgagattctg agtcaatgct tgatgtactc gtggcttcac ttacactagt
     6661 cgaaatggtt gaggtacctg aagatgttac atgatactta ccggtagcgt gcttattgaa
     6721 atcttcatcc gaagttgtca ttgaacttac ccaactcttt gtggatgtgg tctgctgtgt
     6781 tacagaactt gatgaagctg tttgcacgga agaatacgag tgtgacgttt cagcacatgt
     6841 ggagcatgca tatgaggatt caaccgttga ggtgcttaca gaagttgcga cgtttttagt
     6901 ggacattttt atagaagatg taatcaaact tgttgtctgc gagggcatgc aagactcgtg
     6961 agtgcagact gtagcgtctg ttgtaaccgt ttcatgcata ctggttaatg tgctagtctc
     7021 agattcagag ctataaggac accatgtcgt atacattgta gtgactccac taacagtggt
     7081 gaaatatgta gacatggaaa atgatgggat agtagtagaa attgaggatt tggtgatata
     7141 ttcagacgaa ctttgtgaac taacagttgt ttcaacagtt gaagaagtca tagaaggagt
     7201 tgcaacattt gacgaagtcg aagggacgga gtaaacaggt gtggatgggc tgtataatga
     7261 cacagacgtt gatgaagatg ctatagagga gccaagggat gaagttgaat cagtaaaagt
     7321 agagctaata gatgttgaac ttggagaagt ggaagccaaa gttggagatg aagaagtcaa
     7381 acttggggat gtagatgttg aatatgaaga agtggaagtg gagcttgcag aagtagattt
     7441 agaacttgga gatgtagatg ttgaacttga tgaggtagat gtagagcttg gagatgtaga
     7501 tgtcgaactt gatgaggtag atgtagaact ttgggatgta gatgtagaac ttgaagatgt
     7561 ggatgtcaaa cttgaagatg tggatgtcga acttggagat gtagatgtcg aacttgatga
     7621 ggtagatgtc gaacttgatg aggtagatgt cgaacttgat gaggtagatg tagagcttgg
     7681 agatgtagat gtagagcttg gagatgtaga tgtcgaactt agggaggtag tagttggatt
     7741 ggacgatgtt gtactcgaaa gtgtagaggt gacaggtgag attatagagg cacttgatgg
     7801 ttcaatggcg ctggatgata ccacggttgt agttcctact tcggataatg agctgatagc
     7861 ggttgtgtga caagtagggc aaatatatga agtgaacttg gaagtcacag ataaggtggt
     7921 ggttggcaac gtcgatgacg aaggacatgc ctcatgggag cagacttctg tagataatgt
     7981 caaagtacta aatctggata gggtagtagc gtacgatatt gaaggactta tttcggcagc
     8041 agatgaagtg gataccgtca atggacacca agtcgtatac aatgtcgtag tgccagcttg
     8101 aacgatagtg gatgtggaga ctagcgcggg tgaaactgta gttgtaacaa ccccatttgc
     8161 atcgtttgtc ttggttatcg tcactagaat cgtttctgga tcagatgcca aggcaatatt
     8221 agttaatccc aacaatattg tgaacaggta ggtaaaatga gcgaaagata atgtcattta
     8281 tattgaattt tcaaaaattc ttactttttt tttggatgga cgcaaagaag tttaataatc
     8341 atattacatg gcattaccac catatacata tccatatcta atcttactta tatgttgtgg
     8401 aaatgtaaag agccccatta tcttagccta aaaaaacctt ctctttggaa ctttcagtaa
     8461 tacgcttaac tgctcattgc tatattgaag ta
//
