LOCUS       sequence-56729-e        7119 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 7119)
  AUTHORS   Chu D, Kazana E, Bellanger N, Singh T, Tuite MF, von der Haar T
  TITLE     Translation elongation can control translation initiation on 
            eukaryotic mRNAs.
  JOURNAL   EMBO J. 2014 Jan 1;33(1):21-34. doi: 10.1002/embj.201385651. Epub 
            2013 Dec 19.
  PUBMED    24357599
REFERENCE   2  (bases 1 to 7119)
  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: 
            "10.1002/embj.201385651"; journalName: "EMBO J"; date: "2014-01-1- 
            1"; volume: "33"; issue: "1"; pages: "21-34"
COMMENT     Sequence Label: pTH738-CENBEVYslow
FEATURES             Location/Qualifiers
     source          1..7119
                     /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"
     protein_bind    2622..2643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     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(2823..3122)
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
     promoter        3564..4208
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     4101..4124
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      4883..5070
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4938..4962)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5135..5154)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5136..5154)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5161..5178)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5161..5177)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5170..5192)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5322..5777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5409..5428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5619..5640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5906..6709)
                     /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(6710..6930)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6950..6969)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7069..7091
                     /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 ttagctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt
     2761 aaccctcact aaagggaaca aaagctggta ccgggccccc cctcgagggc atgcgaagga
     2821 aaatgagaaa tatcgaggga gacgattcag aggagcagga caaactataa ccgactgttt
     2881 gttggaggat gccgtacata acgaacactg ctgaagctac catgtctaca gtttagagga
     2941 atgggtacaa ctcacaggcg agggatggtg ttcactcgtg ctagcaaacg cggtgggagc
     3001 aaaaagtaga atattatctt ttattcgtga aacttcgaac actgtcatct aaagatgcta
     3061 tatactaata taggcatact tgataatgaa aactataaat cgtaaagaca taagagatcc
     3121 gcttatttag agtgtcgaat tcgagctcgg tacccgggga gttgattgta gcttggtata
     3181 gcttgaaata ttgtgcagaa aaagaaacaa ggaagaaagg gaacgagaac aatgacgagg
     3241 aaacaaaaga ttaataattg caggtctatt tatacttgat agcaagacag caaacttttt
     3301 ttatttcaaa ttcaagtaac tggaaggaag gccgtatacc gttgctcatt agagagtagt
     3361 gtgcgtgaat gaaggaagga aaaagtttcg tgtgttcgaa gatacccctc atcagctctg
     3421 gaacaacgac atctgttggt gctgtctttg tcgttaattt tttcctttag tgtcttccat
     3481 catttttttt gtcattgcgg atatggtgag acaacaacgg gggagagaga aaagaaaaaa
     3541 aaagaaaaga atccgggagt ttatcattat caatactgcc atttcaaaga atacgtaaat
     3601 aattaatagt agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc
     3661 tgtaacccgt acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt
     3721 gtctgggtga acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc
     3781 tggcatccag aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca
     3841 gttcataggt ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa
     3901 cgggcacaac ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat
     3961 tgacccacgc atgtatctat ctcattttct tacaccttct attaccttct gctctctctg
     4021 atttggaaaa agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg
     4081 actaataagt atataaagaa cggtaggtat tgattgtaat tctgtaaatc tatttcttaa
     4141 acttcttaaa ttctactttt atagttagtc ttttttttag ttttaaaaca ccaagaactt
     4201 agtttcgacg gatctagttt cactagcgaa ttatacaact caccagccac acagctcact
     4261 catctacttc gcaatcaaaa caaaatattt tattttagtt cagtttatta agttattatc
     4321 agtatcgtat taaaaaatta aagatcattg aaaaatggct tgctaaaccg attatatttt
     4381 gtttttaaag tagattatta ttagaaaatt attaagagaa ttctgtgtta aatttattga
     4441 aagagaaaat ttattttccc ttattaatta aagtccttta ctttttttga aaactgtcag
     4501 ttttttgaag agttatttgt tttgttacca attgctatca ggtacccgta gaattttatt
     4561 caagatcttt ccgtaacggt tacctttctg tcaaattatc caggtttact cgccaataaa
     4621 aatttcccta tactatcatt aattaaatca ttattattac taaagttttg tttaccaatt
     4681 tgtctgctca agaaaataaa ttaaatacaa ataaaggatc ctctagagtc gacctgcagg
     4741 catgcaagct ttggacttct tcgccagagg tttggtcaag tctccaatca aggttgtcgg
     4801 cttgtctacc ttgccagaaa tttacgaaaa gatggaaaag ggtcaaatcg ttggtagata
     4861 cgttgttgac acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata
     4921 agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa
     4981 attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga
     5041 ggtcgctctt attgaccaca cctctaccgg catgccggtc gaaattcccc taccctatga
     5101 aaagcggccg ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt attacaattc
     5161 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5221 ccttgcagca catcccccct tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5281 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc gacgcgccct gtagcggcgc
     5341 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5401 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5461 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5521 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5581 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5641 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5701 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5761 attaacgttt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5821 caccgcaggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     5881 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     5941 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     6001 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     6061 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     6121 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     6181 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     6241 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     6301 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     6361 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     6421 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     6481 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     6541 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     6601 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     6661 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     6721 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     6781 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     6841 ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag
     6901 aataaaaaaa aaatgatgaa ttgaattgaa aagcgtggtg cactctcagt acaatctgct
     6961 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     7021 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     7081 tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
