LOCUS       sequence-234689-        6482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ATP3-67 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6482)
  AUTHORS   Vowinckel J, Hartl J, Marx H, Kerick M, Runggatscher K, Keller MA, 
            Mulleder M, Day J, Weber M, Rinnerthaler M, Yu JSL, Aulakh SK, 
            Lehmann A, Mattanovich D, Timmermann B, Zhang N, Dunn CD, MacRae JI,
            Breitenbach M, Ralser M
  TITLE     The metabolic growth limitations of petite cells lacking the 
            mitochondrial genome.
  JOURNAL   Nat Metab. 2021 Nov;3(11):1521-1535. doi: 
            10.1038/s42255-021-00477-6. Epub 2021 Nov 18.
  PUBMED    34799698
REFERENCE   2  (bases 1 to 6482)
  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/s42255-021-00477-6"; journalName: "Nat Metab"; date: 
            "2021-11"; volume: "3"; issue: "11"; pages: "1521-1535"
COMMENT     Sequence Label: pRS313_ATP3mut
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(38..56)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(77..93)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(77..93)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(90..112)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    101..117
                     /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(125..131,132..149,150..155))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    170..191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(308..325)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(479..1067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(559..578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1238..2029,2030..2098))
                     /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     1861..1880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2099..2203)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2233..2736
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2784..2802
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2840..2862)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2962..2981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3128..3315
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3316..3978
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(4238..4693)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4375..4396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4587..4606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4819..4841
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4833..4850
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4834..4850
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4857..4876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4857..4875
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4908..4924
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4908..4924
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 tcgacctcga gggggggccc ggtaccagct tttgttccct ttagtgaggg ttaatttcga
       61 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
      121 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
      181 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      241 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
      301 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      361 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
      421 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      481 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      541 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      601 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      661 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      721 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      781 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      841 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      901 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      961 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1021 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1081 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1141 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1201 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1261 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1321 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1381 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1441 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1501 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1561 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1621 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1681 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1741 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1801 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1861 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1921 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1981 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2041 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2101 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2161 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2221 tgccacctga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt taatgatgga
     2281 ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg gattttagaa
     2341 agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac aaaggtttaa
     2401 aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa agcagattaa
     2461 atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg
     2521 tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag gaagagcaag
     2581 ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa
     2641 actatttttt ctttaatttc tttttttact ttctattttt aatttatata tttatattaa
     2701 aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggaccctaa gaaaccatta
     2761 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     2821 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     2881 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     2941 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccataaat
     3001 tcccgtttta agagcttggt gagcgctagg agtcactgcc aggtatcgtt tgaacacggc
     3061 attagtcagg gaagtcataa cacagtcctt tcccgcaatt ttctttttct attactcttg
     3121 gcctcctcta gtacactcta tattttttta tgcctcggta atgattttca tttttttttt
     3181 tccacctagc ggatgactct ttttttttct tagcgattgg cattatcaca taatgaatta
     3241 tacattatat aaagtaatgt gatttcttcg aagaatatac taaaaaatga gcaggcaaga
     3301 taaacgaagg caaagatgac agagcagaaa gccctagtaa agcgtattac aaatgaaacc
     3361 aagattcaga ttgcgatctc tttaaagggt ggtcccctag cgatagagca ctcgatcttc
     3421 ccagaaaaag aggcagaagc agtagcagaa caggccacac aatcgcaagt gattaacgtc
     3481 cacacaggta tagggtttct ggaccatatg atacatgctc tggccaagca ttccggctgg
     3541 tcgctaatcg ttgagtgcat tggtgactta cacatagacg accatcacac cactgaagac
     3601 tgcgggattg ctctcggtca agcttttaaa gaggccctag gggccgtgcg tggagtaaaa
     3661 aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt ggtagatctt
     3721 tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc
     3781 tcttgcgaga tgatcccgca ttttcttgaa agctttgcag aggctagcag aattaccctc
     3841 cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc gttcaaggct
     3901 cttgcggttg ccataagaga agccacctcg cccaatggta ccaacgatgt tccctccacc
     3961 aaaggtgttc ttatgtagtg acaccgatta tttaaagctg cagcatacga tatatataca
     4021 tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt atgatactga
     4081 agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc gctttccttt
     4141 tttctttttg ctttttcttt ttttttctct tgaactcgac ggatctatgc ggtgtgaaat
     4201 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     4261 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4321 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4381 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4441 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4501 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4561 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4621 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4681 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4741 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     4801 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
     4861 acgactcact atagggcgaa ttggagctcc accgcggtgg cggccgctct agaactagtg
     4921 gatcctctct aaaagccgtg tcgcagtaac tggtgacgct aaagttctcc ggcgtttttt
     4981 tatgtatgag ctgggcatca agagcttttg agattttcca agtagtaact catctttctg
     5041 agtgtgctat caaatacata ctaaggagaa taaactcttg ttattacgta ttcttcatcc
     5101 ttatgggtag agagcgcact gttttagtac attttctaga cgtcgaaacg tagagcaatt
     5161 gtcgataaaa caaaaaaaaa gtaagaagat atatgaatag gagctgtcgc tagaactagt
     5221 aagtatatga tggagatata ataagtgaat tattcgatat ttaatgaacg ttctcattta
     5281 tttggaagaa atgtttatca cgtgatggag aaccaatgag cggcgagtaa ctacgcgagg
     5341 aacccggacc gcaataacga ttaaagaagg cccggaaggg agatgcttaa atgattatca
     5401 ctcagttaaa aaagacaaat aagaaactat tgagactgaa ccgttttggt taatttcagg
     5461 tggaaacaat tgaagacgag cagtaaacat tattttattt agtagtcatg ttgtcaagaa
     5521 ttgtatcaaa caatgcaaca cgctccgtaa tgtgccacca agcgcaagtg ggtattcttt
     5581 ataagactaa cccagtgaga acttatgcta ctttgaaaga agtggaaatg cgtttgaaat
     5641 ctatcaaaaa tattgagaag atcacaaaaa ctatgaagat tgttgcatct acaagattga
     5701 gtaaagctga aaaggctaaa atttccgcaa agaagatgga tgaagcagag cagttgtttt
     5761 acaagaacgc cgaaaccaaa aatttggatg ttgaggctac tgaaacaggt gctcctaaag
     5821 agttgattgt tgctatcacc tctgataagg ggttgtgtgg ttctatccac tctcaattgg
     5881 ctaaagctgt gagaagacat ttgaatgatc aaccaaacgc cgatatagtc actattggtg
     5941 ataaaattaa aatgcagcta ttgagaaccc atcctaacaa cattaaattg tctattaatg
     6001 gaattggtaa agatgcccca actttccaag aatctgcttt gattgccgat aagttattga
     6061 gtgtcatgaa ggccggcact tacccaaaga tttccatttt ctacaatgac ccagtgtctt
     6121 ccctatcttt tgaaccatct gaaaaaccga tctttaacgc caagaccatt gaacaatccc
     6181 catcattcgg caaatttgag atcgacacgg acgcaaacgt tccaagagat ttgtttgaat
     6241 atactttggc taaccaaatg ttgacagcaa tggctcaagg ttatgctgct gaaatttccg
     6301 ccagaagaaa cgctatggat aacgcttcca agaatgccgg tgatatgatc aatcgttact
     6361 ctatcttgta caacagaaca agacaagctg tcattactaa tgaactggtt gatattaata
     6421 ctggtccttc ctctttggga tgactcgagt taatcactag tgaattcgcg gccgcctgca
     6481 gg
//
