LOCUS       sequence-74794-e        9115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human MCU in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9115)
  AUTHORS   Sancak Y, Markhard AL, Kitami T, Kovacs-Bogdan E, Kamer KJ, Udeshi 
            ND, Carr SA, Chaudhuri D, Clapham DE, Li AA, Calvo SE, Goldberger O,
            Mootha VK
  TITLE     EMRE is an Essential Component of the Mitochondrial Calcium 
            Uniporter Complex.
  JOURNAL   Science. 2013 Nov 14.
  PUBMED    24231807
REFERENCE   2  (bases 1 to 9115)
  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: "Science. 
            2013 Nov 14."
COMMENT     Sequence Label: pLYS5-MCU-Flag
FEATURES             Location/Qualifiers
     source          1..9115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     285..305
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1452..1475
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1526..1643
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2135..2154
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     LTR             3623..3856
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3928..4049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3965..3984)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4019..4038
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4089..4224
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4244..4263)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4245..4263)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4273..4290)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4273..4289)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4282..4304)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4431..4886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4518..4537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4728..4749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4912..5016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5017..5085,5086..5877)
                     /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(5235..5254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6048..6636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6537..6556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6790..6807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6924..6945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6960..6965,6966..6983,6984..6990)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6998..7014
                     /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     7003..7025
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7022..7038
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7022..7038
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7057..7077
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7059..7077
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7105..7331
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7332..7512
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7559..7684
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8177..8410
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8595..8639
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     8937..8957
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcatggcggc cgccgcaggt agatcgctcc tgctgctcct ctcctctcgg ggcggcggcg
      421 gcgggggcgc cggcggctgc ggggcgctga ctgccggctg cttccctggg ctgggcgtca
      481 gccgccaccg gcagcagcag caccaccgga cggtacacca gaggatcgct tcctggcaga
      541 atttgggagc tgtttattgc agcactgttg tgccctctga tgatgttaca gtggtttatc
      601 aaaatgggtt acctgtgata tctgtgaggc taccatcccg gcgtgaacgc tgtcagttca
      661 cactcaagcc tatctctgac tctgttggtg tatttttacg acaactgcaa gaagaggatc
      721 ggggaattga cagagttgct atctattcac cagatggtgt tcgcgttgct gcttcaacag
      781 gaatagacct cctcctcctt gatgacttta agctggtcat taatgactta acataccacg
      841 tacgaccacc aaaaagagac ctcttaagtc atgaaaatgc agcaacgctg aatgatgtaa
      901 agacattggt ccagcaacta tacaccacac tgtgcattga gcagcaccag ttaaacaagg
      961 aaagggagct tattgaaaga ctagaggatc tcaaagagca gctggctccc ctggaaaagg
     1021 tacgaattga gattagcaga aaagctgaga agaggaccac tttggtgcta tggggtggcc
     1081 ttgcctacat ggccacacag tttggcattt tggcccggct tacctggtgg gaatattcct
     1141 gggacatcat ggagccagta acatacttca tcacttatgg aagtgccatg gcaatgtatg
     1201 catattttgt aatgacacgc caggaatatg tttatccaga agccagagac agacaatact
     1261 tactattttt ccataaagga gccaaaaagt cacgttttga cctagagaaa tacaatcaac
     1321 tcaaggatgc aattgctcag gcagaaatgg accttaagag actgagagac ccattacaag
     1381 tacatctgcc tctccgacaa attggtgaaa aagattctag aggtggatct ggtggatctg
     1441 gtggatctat ggattacaag gatgacgatg acaagtaagg taccgaattc tcgacctcga
     1501 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     1561 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     1621 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc
     1681 cgcggctcga gggggttggg gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga
     1741 cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca
     1801 ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt gggccccccg
     1861 gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg
     1921 acgtgacaaa cggaagccgc acgtctcact agtctcgtgc agatggacag caccgctgag
     1981 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc
     2041 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc
     2101 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     2161 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcat cccgccacca
     2221 tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     2281 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     2341 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     2401 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     2461 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     2521 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     2581 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     2641 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     2701 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     2761 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     2821 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     2881 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     2941 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     3001 ggctccgggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     3061 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     3121 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     3181 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     3241 gaatagagta gatgccgacc gaacaagagc tgatttcgag aacgcctcag ccagcaactc
     3301 gcgcgagcct agcaaggcaa atgcgagaga acggccttac gcttggtggc acagttctcg
     3361 tccacagttc gctaagctcg ctcggctggg tcgcgggagg gccggtcgca gtgattcagg
     3421 cccttctgga ttgtgttggt ccccagggca cgattgtcat gcccacgcac tcgggtgatc
     3481 tgactgatcc cgcagattgg agatcgccgc ccgtgcctgc cgattgggtg cagatccgtc
     3541 gacctgcagc caagctcttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     3601 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     3661 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     3721 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     3781 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     3841 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     3901 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     3961 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4021 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     4081 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4141 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4201 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     4261 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     4321 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     4381 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     4441 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4501 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4561 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4621 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4681 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4741 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4801 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4861 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     4921 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4981 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5041 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5101 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5161 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5221 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5281 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5341 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5401 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5461 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5521 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5581 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5641 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5701 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5761 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5821 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5881 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5941 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6001 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6061 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6121 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6181 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6241 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6301 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6361 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6421 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6481 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6541 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6601 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6661 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6721 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6781 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6841 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6901 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6961 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7021 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     7081 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
     7141 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
     7201 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
     7261 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
     7321 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     7381 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     7441 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     7501 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     7561 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     7621 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     7681 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     7741 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     7801 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     7861 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     7921 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     7981 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
     8041 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     8101 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     8161 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     8221 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     8281 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     8341 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     8401 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     8461 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     8521 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     8581 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     8641 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     8701 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     8761 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     8821 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     8881 acggatctcg acggtatcga tcacgagact agcctcgagc ggccgccccc ttcaccgagg
     8941 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     9001 ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     9061 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatca
//
