LOCUS       sequence-117119-        8452 bp    DNA     circular SYN 14-APR-2025
DEFINITION  DdMCU-Flag in pACT2 vector human codon optimised.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8452)
  AUTHORS   Kovacs-Bogdan E, Sancak Y, Kamer KJ, Plovanich M, Jambhekar A, Huber
            RJ, Myre MA, Blower MD, Mootha VK
  TITLE     Reconstitution of the mitochondrial calcium uniporter in yeast.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Jun 17;111(24):8985-90. doi: 
            10.1073/pnas.1400514111. Epub 2014 Jun 2.
  PUBMED    24889638
REFERENCE   2  (bases 1 to 8452)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1400514111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-06-17- 17"; volume: "111"; issue: "24"; pages: "8985-90"
COMMENT     Sequence Label: DdMCU
FEATURES             Location/Qualifiers
     source          1..8452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1936..2076
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2002..2021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2162..2184
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2213..2614
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3485..3508
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      3649..3836
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3704..3728)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(3854..3887)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3965..3998)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(4042..4075)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(4153..4186)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(4230..4263)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(4341..4374)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4883..5977)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5990..6394)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      6871..8213
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7834..7881)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     1981 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     2041 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     2101 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2161 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcaa
     2221 cttcttttct ttttttttct tttctctctc ccccgttgtt gtctcaccat atccgcaatg
     2281 acaaaaaaat gatggaagac actaaaggaa aaaattaacg acaaagacag caccaacaga
     2341 tgtcgttgtt ccagagctga tgaggggtat ctcgaagcac acgaaacttt ttccttcctt
     2401 cattcacgca cactactctc taatgagcaa cggtatacgg ccttccttcc agttacttga
     2461 atttgaaata aaaaaaagtt tgctgtcttg ctatcaagta taaatagacc tgcaattatt
     2521 aatcttttgt ttcctcgtca ttgttctcgt tccctttctt ccttgtttct ttttctgcac
     2581 aatatttcaa gctataccaa gcatacaatc aactccaagc ttgatgaata gcttcgtgat
     2641 caggaacggc ttcggcctcg tcaggacctt caacaccagg ctgtttacca cctccaccca
     2701 gaacctggag ggcgagctca aaaccatcct cggccaggcc aaggtcagca agctgcagga
     2761 aaagctgaag ctggatccca ggtccaagat caccttcaac gacttcaagg gcatcgccaa
     2821 ggaggtgggc atcgaggaga aggaaatcaa cagcgtcagc aacgccctgg ctcaatccgg
     2881 ctccatcatc tacctgccta actccctgaa cgagaacctg aagacctccg tgttcaccaa
     2941 gcccgcccat atctaccagt ccctcgaaca catcctggac atcgagaaca agggcgtggg
     3001 cctgaataag ctgatcgagt ccaagaaatc cgagatcaat agcctgaggc agaagatcca
     3061 gcccctggag gagaagaagc aggtgatcga cagaaaggcc cacaggaggg ccaccgctat
     3121 catctggacc ggactcggct attgcttcgc ccaggccgcc attctggcta ggctcacctg
     3181 gtgggacctc tcctgggaca tcatcgagcc cgtgagctac ttcctgacct tcggctccgt
     3241 cctgatcggc tacacctact tcaccatgac caagaccgag ttcacctacg aggctctcaa
     3301 ccataggctg ttcagcaaga ggcaggacaa gctgttcaaa agaaacaact tccctaagga
     3361 ggactacgag aacctcgtgc aggccatcga caagaaggag aaggagctca aggagctcga
     3421 gctcgccaca aagtacgatc acacccaccc cgggggtgga tctggtggat ctggtggatc
     3481 tatggattac aaggatgacg atgacaagta aaagctttgg acttcttcgc cagaggtttg
     3541 gtcaagtctc caatcaaggt tgtcggcttg tctaccttgc cagaaattta cgaaaagatg
     3601 gaaaagggtc aaatcgttgg tagatacgtt gttgacactt ctaaataagc gaatttctta
     3661 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
     3721 agtgactctt aggttttaaa acgaaaattc ttgttcttga gtaactcttt cctgtaggtc
     3781 aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg
     3841 tgcgagggac ctaataactt cgtatagcat acattatacg aagttatatt aagggttccg
     3901 gatcgcggcc gctcgacctg cagccaagct agcttggctg gacgtaaact cctcttcaga
     3961 cctaataact tcgtatagca tacattatac gaagttatat taagggttat tgaatatgat
     4021 cggaattggt cgagggacct aataacttcg tatagcatac attatacgaa gttatattaa
     4081 gggttccgga tcgcggccgc tcgacctgca gccaagctag cttggctgga cgtaaactcc
     4141 tcttcagacc taataacttc gtatagcata cattatacga agttatatta agggttattg
     4201 aatatgatcg gaattggtcg agggacctaa taacttcgta tagcatacat tatacgaagt
     4261 tatattaagg gttccggatc gcggccgctc gacctgcagc caagctagct tggctggacg
     4321 taaactcctc ttcagaccta ataacttcgt atagcataca ttatacgaag ttatattaag
     4381 ggttattgaa tatgatcgga attggtcgac tacgtcgtta aggccgtttc tgacagagta
     4441 aaattcttga gggaactttc accattatgg gaaatggttc aagaaggtat tgacttaaac
     4501 tccatcaaat ggtcaggtca ttgagtgttt tttatttgtt gtattttttt ttttttagag
     4561 aaaatcctcc aatatataaa ttaggaatca tagtttcatg attttctgtt acacctaact
     4621 ttttgtgtgg tgccctcctc cttgtcaata ttaatgttaa agtgcaattc tttttcctta
     4681 tcacgttgag ccattagtat caatttgctt acctgtattc ctttacatcc tcctttttct
     4741 ccttcttgat aaatgtatgt agattgcgta tatagtttcg tctaccctat gaacatattc
     4801 cattttgtaa tttcgtgtcg tttctattat gaatttcatt tataaagttt atgtacaaat
     4861 atcataaaaa aagagaatct ttttaagcaa ggattttctt aacttcttcg gcgacagcat
     4921 caccgacttc ggtggtactg ttggaaccac ctaaatcacc agttctgata cctgcatcca
     4981 aaaccttttt aactgcatct tcaatggcct taccttcttc aggcaagttc aatgacaatt
     5041 tcaacatcat tgcagcagac aagatagtgg cgatagggtc aaccttattc tttggcaaat
     5101 ctggagcaga accgtggcat ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag
     5161 aggccaagga cgcagatggc aacaaaccca aggaacctgg gataacggag gcttcatcgg
     5221 agatgatatc accaaacatg ttgctggtga ttataatacc atttaggtgg gttgggttct
     5281 taactaggat catggcggca gaatcaatca attgatgttg aaccttcaat gtaggaaatt
     5341 cgttcttgat ggtttcctcc acagtttttc tccataatct tgaagaggcc aaaacattag
     5401 ctttatccaa ggaccaaata ggcaatggtg gctcatgttg tagggccatg aaagcggcca
     5461 ttcttgtgat tctttgcact tctggaacgg tgtattgttc actatcccaa gcgacaccat
     5521 caccatcgtc ttcctttctc ttaccaaagt aaatacctcc cactaattct ctgacaacaa
     5581 cgaagtcagt acctttagca aattgtggct tgattggaga taagtctaaa agagagtcgg
     5641 atgcaaagtt acatggtctt aagttggcgt acaattgaag ttctttacgg atttttagta
     5701 aaccttgttc aggtctaaca ctaccggtac cccatttagg accacccaca gcacctaaca
     5761 aaacggcatc aaccttcttg gaggcttcca gcgcctcatc tggaagtggg acacctgtag
     5821 catcgatagc agcaccacca attaaatgat tttcgaaatc gaacttgaca ttggaacgaa
     5881 catcagaaat agctttaaga accttaatgg cttcggctgt gatttcttga ccaacgtggt
     5941 cacctggcaa aacgacgatc ttcttagggg cagacatagg ggcagacatt agaatggtat
     6001 atccttgaaa tatatatata tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa
     6061 gacgattgct aaccacctat tggaaaaaac aataggtcct taaataatat tgtcaacttc
     6121 aagtattgtg atgcaagcat ttagtcatga acgcttctct attctatatg aaaagccggt
     6181 tccggcctct cacctttcct ttttctccca atttttcagt tgaaaaaggt atatgcgtca
     6241 ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg
     6301 cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg ttgctaagag attcgaactc
     6361 ttgcatctta cgatacctga gtattcccac agttccttac cactcttttg ttactctatt
     6421 gatccagctc agcaaaggca gtgtgatcta agattctatc ttcgcgatgt agtaaaacta
     6481 gctagaccga gaaagagact agaaatgcaa aaggcacttc tacaatggct gccatcatta
     6541 ttatccgatg tgacgctgca gcttctcaat gatattcgaa tacgctttga ggagatacag
     6601 cctaatatcc gacaaactgt tttacagatt tacgatcgta cttgttaccc atcattgaat
     6661 tttgaacatc cgaacctggg agttttccct gaaacagata gtatatttga acctgtataa
     6721 taatatatag tctagcgctt tacggaagac aatgtatgta tttcggttcc tggagaaact
     6781 attgcatcta ttgcataggt aatcttgcac gtcgcatccc cggttcattt tctgcgtttc
     6841 catcttgcac ttcaatagca tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa
     6901 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     6961 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     7021 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     7081 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     7141 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     7201 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     7261 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     7321 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     7381 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     7441 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     7501 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     7561 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     7621 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     7681 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     7741 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     7801 cttcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagagaatag
     7861 gaacttcgga ataggaactt caaagcgttt ccgaaaacga gcgcttccga aaatgcaacg
     7921 cgagctgcgc acatacagct cactgttcac gtcgcaccta tatctgcgtg ttgcctgtat
     7981 atatatatac atgagaagaa cggcatagtg cgtgtttatg cttaaatgcg tacttatatg
     8041 cgtctattta tgtaggatga aaggtagtct agtacctcct gtgatattat cccattccat
     8101 gcggggtatc gtatgcttcc ttcagcacta ccctttagct gttctatatg ctgccactcc
     8161 tcaattggat tagtctcatc cttcaatgct atcatttcct ttgatattgg atcatatgca
     8221 tagtaccgag aaactagtgc gaagtagtga tcaggtattg ctgttatctg atgagtatac
     8281 gttgtcctgg ccacggcaga agcacgctta tcgctccaat ttcccacaac attagtcaac
     8341 tccgttaggc ccttcattga aagaaatgag gtcatcaaat gtcttccaat gtgagatttt
     8401 gggccatttt ttatagcaaa gattgaataa ggcgcatttt tcttcaaagc tc
//
