LOCUS       sequence-172870-        6115 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 6115)
  AUTHORS   True HL, Berlin I, Lindquist SL
  TITLE     Epigenetic regulation of translation reveals hidden genetic 
            variation to produce complex traits.
  JOURNAL   Nature 2004 Sep 9;431(7005):184-7. Epub 2004 Aug 15.
  PUBMED    15311209
REFERENCE   2  (bases 1 to 6115)
  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"; journalName: "Nature"; 
            date: "2004-09-9"; volume: "431(7005)"; pages: "184-7. Epub 2004 Aug
            15"
COMMENT     Sequence Label: pTEF-Sup35C
FEATURES             Location/Qualifiers
     source          1..6115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     356..375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     609..626
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    743..764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(779..784,785..802,803..809)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    817..833
                     /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     822..844
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     841..857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     841..857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        878..896
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     924..940
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     925..941
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1037..1435
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     1439..1455
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1439..1455
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2769..2788)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2770..2788)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2795..2812)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2795..2811)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2804..2826)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2952..3407
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3039..3058)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3249..3270
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3538..4341)
                     /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(4342..4557)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4581..4600)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4700..4722
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4760..4778)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4846..4950
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4951..5019,5020..5811)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5169..5188)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5982..6115,1..455)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
       61 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      121 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      181 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      241 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      301 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      361 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      421 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      481 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      541 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      601 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      661 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      721 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      781 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      841 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
      901 aaaagctggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt
      961 cctcgagaag atatccatca tattaccatt gtaatactgc caggaataat aggaccaacc
     1021 aacaggcaaa cagtttcata gcttcaaaat gtttctactc cttttttact cttccagatt
     1081 ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt
     1141 tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa
     1201 aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt
     1261 ttctttttct tgaaaatttt ttttttgatt tttttctctt tcgatgacct cccattgata
     1321 tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt
     1381 acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagttttc
     1441 tagaactagt ggatccatgt ttggtggtaa agatcacgtt tctttaattt tcatgggtca
     1501 tgttgatgcc ggtaaatcta ctatgggtgg taatctacta tacttgactg gctctgtgga
     1561 taagagaact attgagaaat atgaaagaga agccaaggat gcaggcagac aaggttggta
     1621 cttgtcatgg gtcatggata ccaacaaaga agaaagaaat gatggtaaga ctatcgaagt
     1681 tggtaaggcc tactttgaaa ctgaaaaaag gcgttatacc atattggatg ctcctggtca
     1741 taaaatgtac gtttccgaga tgatcggtgg tgcttctcaa gctgatgttg gtgttttggt
     1801 catttccgcc agaaagggtg agtacgaaac cggttttgag agaggtggtc aaactcgtga
     1861 acacgcccta ttggccaaga cccaaggtgt taataagatg gttgtcgtcg taaataagat
     1921 ggatgaccca accgttaact ggtctaagga acgttacgac caatgtgtga gtaatgtcag
     1981 caatttcttg agagcaattg gttacaacat taagacagac gttgtattta tgccagtatc
     2041 cggctacagt ggtgcaaatt tgaaagatca cgtagatcca aaagaatgcc catggtacac
     2101 cggcccaact ctgttagaat atctggatac aatgaaccac gtcgaccgtc acatcaatgc
     2161 tccattcatg ttgcctattg ccgctaagat gaaggatcta ggtaccatcg ttgaaggtaa
     2221 aattgaatcc ggtcatatca aaaagggtca atccacccta ctgatgccta acaaaaccgc
     2281 tgtggaaatt caaaatattt acaacgaaac tgaaaatgaa gttgatatgg ctatgtgtgg
     2341 tgagcaagtt aaactaagaa tcaaaggtgt tgaagaagaa gacatttcac caggttttgt
     2401 actaacatcg ccaaagaacc ctatcaagag tgttaccaag tttgtagctc aaattgctat
     2461 tgtagaatta aaatctatca tagcagccgg tttttcatgt gttatgcatg ttcatacagc
     2521 aattgaagag gtacatattg ttaagttatt gcacaaatta gaaaagggta ccaaccgtaa
     2581 gtcaaagaaa ccacctgctt ttgctaagaa gggtatgaag gtcatcgctg ttttagaaac
     2641 tgaagctcca gtttgtgtgg aaacttacca agattaccct caattaggta gattcacttt
     2701 gagagatcaa ggtaccacaa tagcaattgg taaaattgtt aaaattgccg agtaagagct
     2761 ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
     2821 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     2881 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     2941 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3001 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3061 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3121 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3181 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3241 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3301 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3361 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat
     3421 tttctcctta cgcatctgtg cggtatttca caccgcatag ggtaataact gatataatta
     3481 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
     3541 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
     3601 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
     3661 tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc
     3721 atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
     3781 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt
     3841 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
     3901 aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
     3961 tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc
     4021 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag
     4081 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
     4141 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
     4201 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
     4261 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
     4321 cttatatgta gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag
     4381 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
     4441 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
     4501 aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag
     4561 gtggtatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4621 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4681 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4741 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4801 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4861 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4921 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4981 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5041 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5101 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5161 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5221 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5281 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5341 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5401 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5461 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5521 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5581 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5641 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5701 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5761 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5821 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5881 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5941 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6001 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6061 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcaga
//
