LOCUS       sequence-48300-e        8054 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8054)
  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 8054)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pTH741-CENslow-RLuc
FEATURES             Location/Qualifiers
     source          1..8054
                     /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"
     CDS             complement(3154..4086)
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(4094..4490)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     promoter        4499..5143
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     5036..5059
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      5818..6005
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5873..5897)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6070..6089)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6071..6089)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6096..6113)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6096..6112)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6105..6127)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6257..6712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6344..6363)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6554..6575
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6841..7644)
                     /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(7645..7865)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7885..7904)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8004..8026
                     /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 ttgataatgg aaactataaa tcgtgaaggc ataggagatc
     3121 cgcttattta gaagtgtcga attcttacga catttgttca tttttgagaa ctcgctcaac
     3181 gaacgatttg atatattttc ccatttcatc aggtgcatct tcttgcgaaa aatgaagacc
     3241 ttttactttg acaaattcag tattaggaaa cttcttggca ccttcaacaa tagcattgga
     3301 aaagaatcct gggtccgatt caataaacat ttttggtaaa tcatcacttg cacgtagata
     3361 agcattataa ttcctaacaa tttgtacaac gtcaggttta ccacctttta ctaacgggat
     3421 ttcacgaggc catgataatg ttggacgacg aacttcacct ttctctttga atggttcaag
     3481 atatgctgca aattcttctg gttctaactt tctcatgatt tttgatggca acatggtttc
     3541 cacgaagaag ttattctcca aaaccatttt ttctccttct tcagatttga tcaacgcaat
     3601 atcttcttca atatcaggcc attcatccca tgattcaatc acatctacta cactttcagc
     3661 gtgaactatt gctttgatct tatcttgatg ctcatagcta taatgaaatg ccaaacaagc
     3721 accccaatca tggccgacaa aaatgatctt ctttggtaaa ttaagaagtt caaaccatgc
     3781 agtaagatat ttgtaatgat caagtaacct ataagaacca ttaccagatt tgcctgattt
     3841 gcccatacca ataaggtctg gtataataca ccgcgctact ggctcaatat gtggcacaac
     3901 atgtcgccat aaataagaag aggccgcgtt accatgtaaa aaaataacag cattttctgc
     3961 atgtttttct gaatcataat aattaataaa tgaatcaaga acattcattt gtttacatct
     4021 ggcccaccac tgcggaccag ttatcatccg tttcctttgt tctggatcat aaactttcga
     4081 agtcatcccg gggagttgat tgtatgcttg gtatagcttg aaatattgtg cagaaaaaga
     4141 aacaaggaag aaagggaacg agaacaatga cgaggaaaca aaagattaat aattgcaggt
     4201 ctatttatac ttgatagcaa agcggcaaac tttttttatt tcaaattcaa gtaactggaa
     4261 ggaaggccgt ataccgttgc tcattagaga gtagtgtgcg tgaatgaagg aaggaaaaag
     4321 tttcgtgtgt tcgaagatac ccctcatcag ctctggaaca acgacatctg ttggtgctgt
     4381 ctttgtcgtt aattttttcc tttagtgtct tccatcattt tttttgtcat tgcggatatg
     4441 gtgagacaac aacgggggag agagaaaaga aaaaaaaaga aaagaatccg ggagtttatc
     4501 attatcaata ctgccatttc aaagaatacg taaataatta atagtagtga ttttcctaac
     4561 tttatttagt caaaaaatta gccttttaat tctgctgtaa cccgtacatg cccaaaatag
     4621 ggggcgggtt acacagaata tataacatcg taggtgtctg ggtgaacagt ttattcctgg
     4681 catccactaa atataatgga gcccgctttt taagctggca tccagaaaaa aaaagaatcc
     4741 cagcaccaaa atattgtttt cttcaccaac catcagttca taggtccatt ctcttagcgc
     4801 aactacagag aacaggggca caaacaggca aaaaacgggc acaacctcaa tggagtgatg
     4861 caacctgcct ggagtaaatg atgacacaag gcaattgacc cacgcatgta tctatctcat
     4921 tttcttacac cttctattac cttctgctct ctctgatttg gaaaaagctg aaaaaaaagg
     4981 ttgaaaccag ttccctgaaa ttattcccct acttgactaa taagtatata aagaacggta
     5041 ggtattgatt gtaattctgt aaatctattt cttaaacttc ttaaattcta cttttatagt
     5101 tagtcttttt tttagtttta aaacaccaag aacttagttt cgacggatct agtttcacta
     5161 gcgaattata caactcacca gccacacagc tcactcatct acttcgcaat caaaacaaaa
     5221 tattttattt tagttcagtt tattaagtta ttatcagtat cgtattaaaa aattaaagat
     5281 cattgaaaaa tggcttgcta aaccgattat attttgtttt taaagtagat tattattaga
     5341 aaattattaa gagaattctg tgttaaattt attgaaagag aaaatttatt ttcccttatt
     5401 aattaaagtc ctttactttt tttgaaaact gtcagttttt tgaagagtta tttgttttgt
     5461 taccaattgc tatcaggtac ccgtagaatt ttattcaaga tctttccgta acggttacct
     5521 ttctgtcaaa ttatccaggt ttactcgcca ataaaaattt ccctatacta tcattaatta
     5581 aatcattatt attactaaag ttttgtttac caatttgtct gctcaagaaa ataaattaaa
     5641 tacaaataaa ggatcctcta gagtcgacct gcaggcatgc aagctttgga cttcttcgcc
     5701 agaggtttgg tcaagtctcc aatcaaggtt gtcggcttgt ctaccttgcc agaaatttac
     5761 gaaaagatgg aaaagggtca aatcgttggt agatacgttg ttgacacttc taaataagcg
     5821 aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac
     5881 aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc
     5941 ctgtaggtca ggttgctttc tcaggtatag catgaggtcg ctcttattga ccacacctct
     6001 accggcatgc cggtcgaaat tcccctaccc tatgaaaagc ggccgccacc gcggtggagc
     6061 tccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     6121 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccccttcgcc
     6181 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     6241 aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6301 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6361 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6421 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6481 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6541 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6601 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6661 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcctgatgc
     6721 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cagggtaata actgatataa
     6781 ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt
     6841 ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc
     6901 accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc
     6961 agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt
     7021 gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc
     7081 catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac
     7141 agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat
     7201 caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat
     7261 acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac
     7321 acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa
     7381 gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga
     7441 aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc
     7501 aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg
     7561 cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg
     7621 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg
     7681 cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata
     7741 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc
     7801 aaaaaaattt caaagaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa
     7861 ttgaaaagcg tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     7921 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     7981 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     8041 accgaaacgc gcga
//
