LOCUS       sequence-13741-e        8960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8960)
  AUTHORS   Merritt GH, Naemi WR, Mugnier P, Webb HM, Tuite MF, von der Haar T
  TITLE     Decoding accuracy in eRF1 mutants and its correlation with 
            pleiotropic quantitative traits in yeast.
  JOURNAL   Nucleic Acids Res. 2010 Sep . 38(16):5479-92.
  PUBMED    20444877
REFERENCE   2  (bases 1 to 8960)
  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: "Nucleic 
            Acids Res. 2010 Sep . 38(16):5479-92."
COMMENT     Sequence Label: pTH353-SUP45wt
FEATURES             Location/Qualifiers
     source          1..8960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(663..1757)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2611..2632)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2823..2842
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3059..3081
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3073..3090
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3074..3090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3097..3116
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3097..3115
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6140..6156)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6141..6157)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6186..6204)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6225..6241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6225..6241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6238..6260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6249..6265
                     /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)."
     promoter        complement(join(6273..6279,6280..6297,6298..6303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6456..6473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6627..7215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7386..8177,8178..8246))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8009..8028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8247..8351)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8388..8891
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8933..8951
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
      241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
      301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
      361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
      421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
      481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
      541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
      601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
      661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
      721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
      781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
      841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
      901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
      961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
     1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
     1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg
     1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
     1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
     1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     2341 atgtaattgt tccgattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
     2401 gaagttctcc tcgagggtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
     2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3001 cagctggcga aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     3061 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     3121 ttggagctcc accgcggtgg cggccgctct agacaatacg aaggaacgat cccgcgtcag
     3181 cagatgtcac aatttgatta tttcattgtc acatgctttt tgactcatcc ttccgcgaaa
     3241 ggagaacttt ttacaatgcc gccggctctg ttagcgtacc cttctacaag cagatagcag
     3301 aacaaacaca tgatatattc aaaaggtgca atgtcagaga acatatattg cgcccctgtc
     3361 ctgtagacat cagtcatttt tcgcgggact tgaatggcgc accattatcc tatacagaaa
     3421 acattaaatt tactgcaaaa ttttgggcaa acgcttggat agactatgaa tccgtcatga
     3481 agacactact tgtaaaatta tatatatcct tttttttcat gtgctagtaa gaattagcaa
     3541 ctactttcat tcatcttcga cgcaacttcg aggaaagcac ctttttactt gctgtagctc
     3601 tattctcttc cccaaccacc ttttccttta ttccaaaatt tttaaaactt tttctgttac
     3661 attatataat cttctgtctg aaatgtttgg atataacgcc tcttgatcca ctttgtatat
     3721 gcgtgctatt tattttcaga tttataaaga gtatgagcgt catttacata aatagctgaa
     3781 gttattcatg gaaaatacga agagcacgta tgtgagccaa cagaacattt gacgtaagac
     3841 tctacaatgt gccaagaact ggacaagtag aggactgaga actttatttc aattcattgc
     3901 tcctttttgg tggcgctacc tttagcgaag gtcaatgatg aatgtgcaca tgctgtcgaa
     3961 accaaaaagc aaattctaac caacttcaaa atgacatagt catctgatat ttctactcat
     4021 tatagatagt atgggagcct tgaaacgaaa agtaagtaaa aagctggata tgagcagtat
     4081 gaggtagacc ttagctacat catttccccc aatagctgct gcaaatatct ggttaaattt
     4141 gtgattccat gaagaggata acagacttgt taaaaagcat cctgtcaaaa tctaattttt
     4201 gaagggcagt attcaattca taatttactt tagcttagat ccttccaatt tatacatggt
     4261 attataacca gatcataaac tacaatctgt cgctaccgca tgtacgaaga atacttaagt
     4321 cacttgctct ctcatcattt gtacattttt cagtaatacc gtttgatagc gccgtcttta
     4381 ttacccggat tattccgttg accctgaatg aaaaattttt tcagaaatcc agtgctaagc
     4441 gtcaaatcaa tgaaatacat cactgtattt ttaactgata tactgttggt gtggccttaa
     4501 cgacaccttt atttcttaat tcatttcggc ttgtctcctt attaagacta cagaaataga
     4561 caaaggaaat acttcaataa tggataacga ggttgaaaaa aatattgaga tctggaaggt
     4621 caagaagttg gtccaatctt tagaaaaagc tagaggtaat ggtacttcta tgatttcctt
     4681 agttattcct cctaagggtc aaattccact gtaccaaaaa atgttaacag atgaatatgg
     4741 tactgcctcg aatattaaat ctagggttaa tcgtctttcc gttttatctg ctatcacttc
     4801 cacccaacaa aagttgaagc tatataatac tttgcccaag aacggtttag ttttatattg
     4861 tggtgatatc atcactgaag atggtaaaga aaaaaaggtc acttttgaca tcgaacctta
     4921 caaacctatc aacacatcct tatatttgtg tgataacaaa tttcatacag aagttctttc
     4981 ggaattgctt caagctgacg acaagttcgg ttttatagtc atggacggtc aaggtacttt
     5041 gtttggttct gtgtccggta atacgagaac tgttttacat aaatttactg tcgatctgcc
     5101 aaaaaagcat ggtagaggtg gtcaatctgc gcttcgtttt gctcgtttaa gagaagaaaa
     5161 aagacataat tatgtgagaa aggtcgccga agttgctgtt caaaatttta ttactaatga
     5221 caaagtcaat gttaagggtt taattttagc tggttctgct gactttaaga ccgatttggc
     5281 taaatctgaa ttattcgatc caagactagc atgtaaggtt atttccatcg tggatgtttc
     5341 ttatggtggt gaaaacggtt tcaaccaggc tatcgaactt tctgccgaag cgttggccaa
     5401 tgtcaagtat gttcaagaaa agaaattatt ggaggcatat tttgacgaaa tttcccagga
     5461 cactggtaaa ttctgttatg gtatagatga tactttaaag gcattggatt taggtgcagt
     5521 cgaaaaatta attgttttcg aaaatttgga aactatcaga tatacattta aagatgccga
     5581 ggataatgag gttataaaat tcgctgaacc agaagccaag gacaagtcgt ttgctattga
     5641 caaagctacc ggccaagaaa tggacgttgt ctccgaagaa cctttaattg aatggctagc
     5701 agctaactac aaaaacttcg gtgctacctt ggaattcatc acagacaaat cttcagaagg
     5761 tgcccaattt gtcacaggtt ttggtggtat tggtgccatg ctgcgttaca aagttaattt
     5821 tgaacaacta gttgatgaat ctgaggatga atattatgac gaagatgaag gatccgacta
     5881 tgatttcatt taaataaata aaagggggag aaaaaaatcg aatcaaaaag aatttaatca
     5941 ctagatgcca gatttaaatt aaattcgctt ttaatttttt gtacaatata atatatactt
     6001 ggtaaacctt ttgctctata ttgagctaat tcctttgttg aaagtacata gtgggtttag
     6061 aggaccgtgt atattacgta gaaaatacag tgaaaggaga gtttctcttc aaaagcctcg
     6121 acggtatcga taagcttatc gataccgtcg acctcgaggg ggggcccggt acccagcttt
     6181 tgttcccttt agtgagggtt aattccgagc ttggcgtaat catggtcata gctgtttcct
     6241 gtgtgaaatt gttatccgct cacaattcca cacaacatag gagccggaag cataaagtgt
     6301 aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg ctcactgccc
     6361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcg gcccccctga cgagcatcac
     6661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6721 ttcccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     6841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7441 tccatagttg cctgactgcc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgaaaa
     7801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8341 ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc acgtgctata
     8401 aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata gaaagtaaaa
     8461 aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact ctagggggat
     8521 cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa tgccgaattg
     8581 tttcatcttg tctgtgtaga agaccacaca cgaaaatcct gtgattttac attttactta
     8641 tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat atatatgtaa
     8701 agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa
     8761 ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa
     8821 cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca
     8881 ggcaagcgat ccgtcctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8941 tatcacgagg ccctttcgtc
//
