LOCUS       sequence-42825-e        7546 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 7546)
  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 May 9. ():.
  PUBMED    21558172
REFERENCE   2  (bases 1 to 7546)
  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"; journalName: "Nucleic 
            Acids Res. 2011 May 9. ():."
COMMENT     Sequence Label: pTH645-RLuc
FEATURES             Location/Qualifiers
     source          1..7546
                     /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..5144
                     /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      5313..5500
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5368..5392)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5562..5581)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5563..5581)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5588..5605)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5588..5604)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5597..5619)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5749..6204
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5836..5855)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6046..6067
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6333..7136)
                     /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(7137..7357)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7377..7396)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7496..7518
                     /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 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 ttttagtttt aaaacaccaa gaacttagtt tcgacggatc ctctagagtc
     5161 gacctgcagg catgcaagct ttggacttct tcgccagagg tttggtcaag tctccaatca
     5221 aggttgtcgg cttgtctacc ttgccagaaa tttacgaaaa gatggaaaag ggtcaaatcg
     5281 ttggtagata cgttgttgac acttctaaat aagcgaattt cttatgattt atgattttta
     5341 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     5401 taaaacgaaa attcttgttc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     5461 tatagcatga ggtcgctctt attgaccaca cctctaccgg catgcccgaa attcccctac
     5521 cctatgaaaa gcggccgcca ccgcggtgga gctccaattc gccctatagt gagtcgtatt
     5581 acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5641 ttaatcgcct tgcagcacat ccccccttcg ccagctggcg taatagcgaa gaggcccgca
     5701 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcgac gcgccctgta
     5761 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5821 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5881 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5941 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6001 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6061 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6121 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6181 acaaaatatt aacgtttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     6241 tatttcacac cgcagggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     6301 agtatacatg catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa
     6361 tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt
     6421 gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg
     6481 gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata
     6541 atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata
     6601 acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat
     6661 agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact
     6721 tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc
     6781 gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta
     6841 ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat
     6901 gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt
     6961 tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta
     7021 cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg
     7081 gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt
     7141 tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg ttaagaatac tgggcaattt
     7201 catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt
     7261 cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaagaaa ccgaaatcaa
     7321 aaaaaagaat aaaaaaaaaa tgatgaattg aattgaaaag cgtggtgcac tctcagtaca
     7381 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     7441 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     7501 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcga
//
