LOCUS       sequence-42826-e        6614 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 6614)
  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 6614)
  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: pTH644-CENBEVY
FEATURES             Location/Qualifiers
     source          1..6614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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     complement(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(2824..3123)
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
     promoter        complement(3162..3558)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     promoter        3567..4212
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     4104..4127
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      4381..4568
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4436..4460)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4630..4649)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4631..4649)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4656..4673)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4656..4672)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4665..4687)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4817..5272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4904..4923)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5114..5135
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5401..6204)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6205..6425)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6445..6464)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6564..6586
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 ggcagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt
     2761 aaccctcact aaagggaaca aaagctgggt accgggcccc ccctcgaggg catgcgaagg
     2821 aaaatgagaa atatcgaggg agacgattca gaggagcagg acaaactata accgactgtt
     2881 tgttggagga tgccgtacat aacgaacact gctgaagcta ccatgtctac agtttagagg
     2941 aatgggtaca actcacaggc gagggatggt gttcactcgt gctagcaaac gcggtgggag
     3001 caaaaagtag aatattatct tttattcgtg aaacttcgaa cactgtcatc taaagatgct
     3061 atatactaat ataggcatac ttgataatga aaactataaa tcgtaaagac ataagagatc
     3121 cgcttattta gaagtgtcga attcgagctc ggtacccggg gagttgattg tatgcttggt
     3181 atagcttgaa atattgtgca gaaaaagaaa caaggaagaa agggaacgag aacaatgacg
     3241 aggaaacaaa agattaataa ttgcaggtct atttatactt gatagcaaag cggcaaactt
     3301 tttttatttc aaattcaagt aactggaagg aaggccgtat accgttgctc attagagagt
     3361 agtgtgcgtg aatgaaggaa ggaaaaagtt tcgtgtgttc gaagataccc ctcatcagct
     3421 ctggaacaac gacatctgtt ggtgctgtct ttgtcgttaa ttttttcctt tagtgtcttc
     3481 catcattttt tttgtcattg cggatatggt gagacaacaa cgggggagag agaaaagaaa
     3541 aaaaaagaaa agaatccggg agtttatcat tatcaatact gccatttcaa agaatacgta
     3601 aataattaat agtagtgatt ttcctaactt tatttagtca aaaaattagc cttttaattc
     3661 tgctgtaacc cgtacatgcc caaaataggg ggcgggttac acagaatata taacatcgta
     3721 ggtgtctggg tgaacagttt attcctggca tccactaaat ataatggagc ccgcttttta
     3781 agctggcatc cagaaaaaaa aagaatccca gcaccaaaat attgttttct tcaccaacca
     3841 tcagttcata ggtccattct cttagcgcaa ctacagagaa caggggcaca aacaggcaaa
     3901 aaacgggcac aacctcaatg gagtgatgca acctgcctgg agtaaatgat gacacaaggc
     3961 aattgaccca cgcatgtatc tatctcattt tcttacacct tctattacct tctgctctct
     4021 ctgatttgga aaaagctgaa aaaaaaggtt gaaaccagtt ccctgaaatt attcccctac
     4081 ttgactaata agtatataaa gaacggtagg tattgattgt aattctgtaa atctatttct
     4141 taaacttctt aaattctact tttatagtta gtcttttttt ttagttttaa aacaccaaga
     4201 acttagtttc gacggatcct ctagagtcga cctgcaggca tgcaagcttt ggacttcttc
     4261 gccagaggtt tggtcaagtc tccaatcaag gttgtcggct tgtctacctt gccagaaatt
     4321 tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa
     4381 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     4441 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttgttctt gagtaactct
     4501 ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc
     4561 tctaccggca tgcccgaaat tcccctaccc tatgaaaagc ggccgccacc gcggtggagc
     4621 tccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     4681 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccccttcgcc
     4741 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4801 aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4861 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4921 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4981 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5041 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5101 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5161 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5221 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcctgatgc
     5281 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cagggtaata actgatataa
     5341 ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt
     5401 ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc
     5461 accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc
     5521 agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt
     5581 gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc
     5641 catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac
     5701 agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat
     5761 caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat
     5821 acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac
     5881 acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa
     5941 gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga
     6001 aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc
     6061 aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg
     6121 cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg
     6181 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg
     6241 cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata
     6301 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc
     6361 aaaaaaattt caaagaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa
     6421 ttgaaaagcg tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6481 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6541 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6601 accgaaacgc gcga
//
