LOCUS       sequence-56700-e        8039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression of AVT1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8039)
  AUTHORS   Hughes AL, Gottschling DE
  TITLE     An early age increase in vacuolar pH limits mitochondrial function 
            and lifespan in yeast.
  JOURNAL   Nature. 2012 Dec 13;492(7428):261-5. doi: 10.1038/nature11654. Epub 
            2012 Nov 21.
  PUBMED    23172144
REFERENCE   2  (bases 1 to 8039)
  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: 
            "10.1038/nature11654"; journalName: "Nature"; date: "2012-12-13- 
            13"; volume: "492"; issue: "7428"; pages: "261-5"
COMMENT     Sequence Label: pAG306-GPD-AVT1 chr I
FEATURES             Location/Qualifiers
     source          1..8039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        190..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /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"
     rep_origin      complement(1343..1798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1480..1501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1692..1711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1950
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1942..1959
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2009..2653
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2547..2570
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     2659..2675
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2659..2675
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2683..2707
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4523..4547)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4580..4596)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4581..4597)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4599..4846
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4615..4633)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(4869..4887)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4908..4924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4908..4924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4921..4943)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4932..4948
                     /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)."
     promoter        complement(join(4956..4962,4963..4980,4981..4986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5139..5156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5310..5898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6069..6860,6861..6929))
                     /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     6692..6711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6930..7034)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8012..8030
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     1981 tagggcgaat tggagctctt cgagtttatc attatcaata ctcgccattt caaagaatac
     2041 gtaaataatt aatagtagtg attttcctaa ctttatttag tcaaaaaatt agccttttaa
     2101 ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat atataacacc
     2161 gaaggtgcct gggtgaacag gtttattcct ggcatccact aaatataatg gagcccgctt
     2221 tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt ttcttcacca
     2281 accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg cacaaacagg
     2341 caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa tgatgacaca
     2401 aggcaattga cccacgcatg tatctatctc attttcttac accttctatt accttctgct
     2461 ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga aattattccc
     2521 ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg taaatctatt
     2581 tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt aaaacaccag
     2641 aacttagttt cgacggattc tagaactagt ggatccccca tcacaagttt gtacaaaaaa
     2701 gcaggctcca ccatgcctga gcaagaacca ttgagtccca atggccgtaa gcgctctgag
     2761 gttcactata tctcaatccc attgaacaga ggttccgcat tctcaccaga tgattctgta
     2821 tcacagtttc agtctgacgg gttcatgaca cgtaggcaat ccatattgga ccatcctgtg
     2881 ggctcattta agggagttaa ttctttaagt cgatttgcga cttcgttgag gagagccaat
     2941 tcgtttcgta atatcgagct aaatgcggat aatgaaagat cctttttcaa agagagtaac
     3001 gatgaaacct acgatccgga tactttagca ccagctttgg acggtagaag attatcagta
     3061 actttaaata atgctggtcg cccgcgcatt actaatttgg ctaacaacga tagagttagt
     3121 acagccagca tggctattca cgatgatgat tatgggtcca tccaaaattc aacaattggg
     3181 gattctgggt caatattacg ccctactgcc tccttaacag aaatgatgag tggtggagcg
     3241 gggagaaggt ttacaaataa tgacatggat tcaattgtag tgaagagagt ggagggtgta
     3301 gatggtaagg tagtaactct tcttgctggt cagtcgaccg ctccacagac catattcaac
     3361 tcaatcaatg tactgattgg tattgggctc ctggcgttac ctcttggttt gaaatacgcc
     3421 ggctgggtta taggacttac tatgcttgca atcttcgcct tagcgacatt ttgtactgct
     3481 gaacttttgt ctaggtgcct agatacagat ccaactttga tatcttacgc ggacttggga
     3541 tatgcagcat ttggtacaaa agggcgcgca ctaatttctg ctcttttcac acttgaccta
     3601 ctgggtagtg gggtgtcctt ggtgattctc ttcggagatt ctttaaatgc tctatttcct
     3661 caatactcca ccactttttt caaaatagtt tcatttttta tcgttacacc cccggttttt
     3721 ataccattga gtgttctatc taatatctcg cttctgggaa tcctttcaac aaccggaact
     3781 gtgctagtca tttgctgctg cggactatac aagtcgtctt ctccaggttc attagttaat
     3841 ccgatggaaa cgagcatgtg gccgatagat ctaaagcatc tctgtttatc tataggttta
     3901 ttgagtgcct gctggggagg acatgcggtt tttcccaact taaagacaga tatgaggcat
     3961 ccagataaat ttaaagactg tttgaaaact acctataaga taacatcggt tacggacata
     4021 ggtactgcag ttattggctt tttgatgttt ggaaatcttg ttaaagatga aattactaag
     4081 aacgttttgc tgacggaggg atatccaaaa ttcgtttatg gtctgatttc agcgcttatg
     4141 actatcattc ctatcgcaaa gaccccattg aacgcaagac ctatcgtttc tgtattggac
     4201 gttctgatga atgtgcaaca catagatgaa gcggccagcg ccataaaaag aagagctgcg
     4261 aaaggccttc aagtattcaa taggatcttt attaatgttg tcttcgtttt gatagctatt
     4321 aatttccctg aatttgataa gattatcgca tttctggggg ccggtttgtg ttttacaatt
     4381 tgcctaattt tgccgtgttg gttctattta agattgtgca agacaacaat caaaccatgg
     4441 gagagagtgg cttgtcacgt aaccatatgt attagtgtcg tgttgtccac attaggtgtt
     4501 ggtgctgcaa ttatttcata ggacccagct ttcttgtaca aagtggtgat gggctgcagg
     4561 aattcgatat caagcttatc gataccgtcg acctcgagtc atgtaattag ttatgtcacg
     4621 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     4681 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     4741 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     4801 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccagc
     4861 ttttgttccc tttagtgagg gttaattccg agcttggcgt aatcatggtc atagctgttt
     4921 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca taggagccgg aagcataaag
     4981 tgtaaagcct ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg
     5041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5101 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tcggcccccc tgacgagcat
     5341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5401 gcgttccccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     5521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6001 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6121 tcatccatag ttgcctgact gcccgtcgtg tagataacta cgatacggga gggcttacca
     6181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6361 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtga
     6481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6661 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7021 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg gggggtgcta attatggcat
     7081 tgattggggc ggtaaaacat tcaaagtcaa tgaaaaggat gctaaattga atgatactga
     7141 agacatttca ttcgcttctc agatcaaaaa cttaaatctg tctatatgag ctgtcccaaa
     7201 gccacttctt acctttcaag aaaacatgga ccttgctttg ccacaaaata attgcctgta
     7261 agatagtttt ttatttgcac attgttatta ctaacctatg ctttgctacg gacgactgct
     7321 tggtaaaaat gcaaaagtat taggatgtct cacaagaaac tgctcgtggt gaaaattttc
     7381 agccgcgacc atcctcccac gccaacaagc cctcttttgc ccatccttct ggaattcgcg
     7441 gtgaaaatca atttgatata taaaattccc tcataatatc taggaacggg atcacatatc
     7501 tcaatgagct aaagcggccg cttcttcgaa tgggaccagc taaagtgatc aaacttcatg
     7561 caggatgttg gacacttttt ctttcaagaa gctccaaata tataaatttt tgcaatatag
     7621 gaattgaaat atcgtacttc cgattcagaa tatctcaact ctactacatc tatatatgac
     7681 tgtactcgcg ttcttggtaa acatcaatta gtaggtgttt ttgcgagcat atgttgagct
     7741 cagaaaatat gagcacaatc ttcgaaacca ttgttttttt gtgcaaaagt caccatcggg
     7801 tttatagtat ggactcttga acgaaccatt ctgatctgta tttctctcat atttactcca
     7861 tttatttttt gctgctcttt gctttacttc aacacgatgc ttttcctaag taatttttta
     7921 ttttattcac aatgctccat aactcttcag gccttaaacg accatccatg gcccctaaga
     7981 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
