LOCUS       sequence-35616-e        3843 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 3843)
  AUTHORS   Baughman JM, Perocchi F, Girgis HS, Plovanich M, Belcher-Timme CA, 
            Sancak Y, Bao XR, Strittmatter L, Goldberger O, Bogorad RL, 
            Koteliansky V, Mootha VK
  TITLE     Integrative genomics identifies MCU as an essential component of the
            mitochondrial calcium uniporter.
  JOURNAL   Nature. 2011 Jun 19;476(7360):341-5. doi: 10.1038/nature10234.
  PUBMED    21685886
REFERENCE   2  (bases 1 to 3843)
  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"; journalName: 
            "Nature"; date: "2011-06-19- 19"; volume: "476"; issue: "7360"; 
            pages: "341-5"
COMMENT     Sequence Label: pDONR223-MCU-E257A
FEATURES             Location/Qualifiers
     source          1..3843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1722..1821)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1838..1857)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1839..1857)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1862..1878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1862..1878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2309..3100
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3193..3781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3682..3701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcatg gcggccgccg caggtagatc gctcctgctg ctcctctcct ctcggggcgg
      721 cggcggcggg ggcgccggcg gctgcggggc gctgactgcc ggctgcttcc ctgggctggg
      781 cgtcagccgc caccggcagc agcagcacca ccggacggta caccagagga tcgcttcctg
      841 gcagaatttg ggagctgttt attgcagcac tgttgtgccc tctgatgatg ttacagtggt
      901 ttatcaaaat gggttacctg tgatatctgt gaggctacca tcccggcgtg aacgctgtca
      961 gttcacactc aagcctatct ctgactctgt tggtgtattt ttacgacaac tgcaagaaga
     1021 ggatcgggga attgacagag ttgctatcta ttcaccagat ggtgttcgcg ttgctgcttc
     1081 aacaggaata gacctcctcc tccttgatga ctttaagctg gtcattaatg acttaacata
     1141 ccacgtacga ccaccaaaaa gagacctctt aagtcatgaa aatgcagcaa cgctgaatga
     1201 tgtaaagaca ttggtccagc aactatacac cacactgtgc attgagcagc accagttaaa
     1261 caaggaaagg gagcttattg aaagactaga ggatctcaaa gagcagctgg ctcccctgga
     1321 aaaggtacga attgagatta gcagaaaagc tgagaagagg accactttgg tgctatgggg
     1381 tggccttgcc tacatggcca cacagtttgg cattttggcc cggcttacct ggtgggccta
     1441 ttcctgggac atcatggagc cagtaacata cttcatcaca tatggaagtg ccatggcaat
     1501 gtatgcatat tttgtaatga cacgccagga atatgtttat ccagaagcca gagacagaca
     1561 atacttacta tttttccata aaggagccaa aaagtcacgt tttgacctag agaaatacaa
     1621 tcaactcaag gatgcaattg ctcaggcaga aatggacctt aagagactga gagacccatt
     1681 acaagtacat ctgcctctcc gacaaattgg tgaaaaagat tgcccaactt tcttgtacaa
     1741 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1801 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1861 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1921 acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa atgcctcgac
     1981 ttcgctgctg cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
     2041 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
     2101 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     2161 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     2221 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     2281 cagtcgccct aaaacaaagt taaacatcat gagggaagcg gtgatcgccg aagtatcgac
     2341 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     2401 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
     2461 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     2521 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     2581 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     2641 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     2701 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     2761 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     2821 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     2881 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     2941 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     3001 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     3061 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc
     3121 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3181 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3241 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3301 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3361 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3421 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3481 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3541 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     3601 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3661 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3721 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3781 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3841 gtt
//
