LOCUS       sequence-13795-e        7447 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 7447)
  AUTHORS   Chu D, Barnes DJ, von der Haar T
  TITLE     The role of tRNA and ribosome competition in coupling the expression
            of different mRNAs in Saccharomyces cerevisiae.
  JOURNAL   Nucleic Acids Res. 2011 Aug;39(15):6705-14. doi: 10.1093/nar/gkr300.
            Epub 2011 May 9.
  PUBMED    21558172
REFERENCE   2  (bases 1 to 7447)
  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: 
            "10.1093/nar/gkr300"; journalName: "Nucleic Acids Res"; date: 
            "2011-08"; volume: "39"; issue: "15"; pages: "6705-14"
COMMENT     Sequence Label: pTH490-tQSRTL
FEATURES             Location/Qualifiers
     source          1..7447
                     /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"
     promoter        complement(4673..4691)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4712..4728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4712..4728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4725..4747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4736..4752
                     /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(4760..4766,4767..4784,4785..4790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4943..4960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5114..5702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5873..6664,6665..6733))
                     /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     6496..6515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6734..6838)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6875..7378
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7420..7438
                     /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 cggccgctcc ggagtaattg acttgatttt gcaaagcttc
     3181 acttaaatgg cgtatcttcc ttttataaat tgcgtttgct ttattccctt acccaacaga
     3241 tgaattggta ctctggccga gtggtctaag gcgtcaggtc gaggtcctga tctcttcgga
     3301 ggcgcgggtt caaaccccgc gggtatcaat atttttttga atctacgcac ttcaataaga
     3361 attaggctgt taggatagca ttttttttct atgtaccaaa agtattcata cgtggctcct
     3421 tgttaccccg gccgctctag aactagtacg cgtgtgattg atcttgtact ggttgtaaat
     3481 gtgctaagtc tgtgactgca gtgaatagga ttgttttaat caacgatgtt atttaaaaaa
     3541 aattaaacga caccagcagg atttgaacca gcgcgggcag agcccaagag atagccgaac
     3601 tttttattcc attcgagtct ctcgccttaa ccactcggcc atagtgccat ttcgatttga
     3661 aattatggtt taaagcggta ggttttatac atgtttctag gcgttttttc acagatccaa
     3721 gacagtagag cagcttggcg taggtaggac tagtggatcc tcatttttaa ggtcattttt
     3781 gctaaatgta tcatcttcaa aatagataaa tctgtaaagt atagcggagc cacaaattta
     3841 gcaggtataa aaattactca gcccttttgg ccaagtggta aggcatcgca ctcgtaatgc
     3901 ggggatcgtg ggttcaattc ccacagaggg catttgaaat tttattttct attttttaat
     3961 ttttatatat aatatatata acgcgacata gctgttatta cctactagta atatcctctc
     4021 ttttttcaaa caggatcccc cgggctgcag gaattcgata tcaagcttaa gatccattgc
     4081 ttgcgatatt gttaaaatag ggaccatctt tcttaaaaag gttccataaa accggaagtt
     4141 ttagtgtaca ctaacaacag aagaaaaagg tcctatagtg tagtggttat cactttcggt
     4201 tctgatccga acaaccccag ttcgaatccg ggtgggacct tcccttcatt tttcgtattt
     4261 tacgaaattt aaaccatcat tttttttttt tgaaccaatg cgagaaaaaa ataaaaagag
     4321 aaaggtaaga aaaagaaacc aagcttatcg ataccgtcga catagacact gtgctggcat
     4381 cgtagcctcc gttctttcta tgcactgact cggaaatatg ctttttcctt aaaaattaga
     4441 atagtgcgct aaaaaattaa aagctcctcc cgggactcga acccggatca cagccaccgg
     4501 aagaatgcat gctaaccatt gcactagagg agcataataa attatcagag atagtaaatt
     4561 cctgctcttt attcatctga cctagtattt attccgaaga caaaactcgg cgactttgat
     4621 gtcgtacttt caggtcgacc tcgagggggg gcccggtacc cagcttttgt tccctttagt
     4681 gagggttaat tccgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4741 atccgctcac aattccacac aacataggag ccggaagcat aaagtgtaaa gcctggggtg
     4801 cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4861 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4921 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4981 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5041 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5101 cgttgctggc gtttttccat aggctcggcc cccctgacga gcatcacaaa aatcgacgct
     5161 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttc ccccctggaa
     5221 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5281 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     5341 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5401 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5461 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5521 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5581 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5641 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5701 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5761 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5821 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5881 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5941 gactgcccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6001 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6061 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6181 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgaaaaaaa gcggttagct
     6301 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6361 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6481 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6541 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6721 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6841 catttccccg aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa
     6901 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag
     6961 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac
     7021 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct
     7081 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat
     7141 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg
     7201 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc
     7261 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt
     7321 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg
     7381 tcctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7441 tttcgtc
//
