LOCUS       Mfn2-Myc.        8419 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    Mfn2-Myc.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8419)
  AUTHORS   Chen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC
  TITLE     Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion 
            and are essential for embryonic development.
  JOURNAL   J Cell Biol. 2003 Jan 20. 160(2):189-200.
  PUBMED    12527753
REFERENCE   2  (bases 1 to 8419)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8419)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol. 2003 Jan 20. 160(2):189-200."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1002..3272
                     /gene="Mfn2"
                     /label=Mfn2
                     /note="Mitofusin-2 from Mus musculus. Accession#: Q80U63"
     CDS             3288..3317
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             3324..3353
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3360..3389
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3408..3437
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3444..3473
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3480..3509
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3528..3557
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3564..3593
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3600..3629
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3648..3677
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3684..3713
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3720..3749
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3768..3797
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3804..3833
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3840..3869
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3903..3932
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3948..3965
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    4015..4239
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4285..4713
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4727..5057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5124..5915
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6092..6225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6262..6278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6286..6302)
                     /label=lac operator
                     /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(6310..6340)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6355..6376)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(6493..6510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6664..7252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7426..8283)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8284..8388)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagtcct tttcatcttt ctgactccag ccatgtccac
      961 gatgcccaac ctgtgaaggt ctcttaccag ctagaaacga gatgtccctg ctcttttctc
     1021 gatgcaactc catcgtcacc gtcaagaagg ataagcgaca catggctgaa gtgaatgctt
     1081 cccctctcaa gcactttgtc actgccaaga aaaagatcaa tggaatcttt gagcagctgg
     1141 gggcctacat ccaagagagc gccagcttcc ttgaagacac ccacaggaac acagaactgg
     1201 acccggttac cacggaagag caggtcctgg acgtcaaagg gtacctgtcc aaggtcaggg
     1261 gtatcagcga agtgctggcc aggcggcaca tgaaggtggc tttttttggc cggacgagca
     1321 atgggaagag caccgtgatc aatgccatgc tctgggacaa agttctgcca tctgggattg
     1381 gtcataccac caattgcttc ctgcgggttg ggggcacaga tggccatgag gccttcctcc
     1441 tcacagaggg ctcagaagag aagaagagtg tcaagactgt gaaccaactg gcccatgccc
     1501 tccatcagga cgagcagttg catgcaggca gcatggtgag tgtgatgtgg cccaactcca
     1561 agtgtccgct cctgaaggat gacctcgtgc tgatggacag ccctgggatc gatgttacca
     1621 cggagctgga cagctggatt gataagtttt gcctggatgc tgatgtgttt gtgctggtgg
     1681 ccaactcaga gtccacgctg atgcagacgg agaagcagtt cttccacaaa gtgagtgaac
     1741 gtctctcccg gcccaacatc ttcatcctga acaaccgctg ggatgcgtct gcctcggagc
     1801 ctgagtacat ggaggaggtg cggcggcagc acatggagcg ctgcaccagc tttctggtgg
     1861 atgagctggg cgtggtggat cgagctcagg ctggggaccg gatcttcttc gtgtctgcca
     1921 aggaggttct cagcgccagg gtccagaaag cccagggcat gccagaagga ggcggcgctc
     1981 tcgcagaagg ttttcaagtg aggatgtttg agtttcagaa tttcgagagg cagtttgagg
     2041 agtgcatttc ccagtctgca gtaaagacca aatttgagca gcacacagtc cgggccaagc
     2101 agattgcaga ggccgttcgt ctcatcatgg attccctgca catcgcagct caggagcagc
     2161 gggtttattg cctagaaatg cgggaagagc ggcaagaccg gctgaggttt attgacaagc
     2221 agctggagct cctggctcaa gactacaagc tgcgaattaa gcagattacg gaggaagtgg
     2281 aaaggcaggt gtccacagcc atggctgaag agatcaggcg cctctctgtg ctagttgacg
     2341 agtaccagat ggacttccac ccatccccag ttgtcctcaa ggtttataag aacgagctgc
     2401 accgccatat agaggaaggt ctgggccgga acctgtctga ccgctgctcc actgccattg
     2461 ccagttcact gcagactatg cagcaggaca tgatagacgg cttgaagccc cttcttcctg
     2521 tatctatgcg gaatcagata gacatgctgg tccctcgaca gtgtttctcc ctcagctatg
     2581 acctgaattg tgacaagctg tgtgctgact ttcaggagga catcgagttc cacttctccc
     2641 ttggatggac tatgctagtg aacaggttcc tgggccccaa gaatagccgc cgggccttgc
     2701 taggctacag tgatcaggtt cagcgtcctc tccctctgac acctgccaac cccagcatgc
     2761 cccccttgcc acagagctcc ctcacccagg aggagctcat ggtctccatg gttactggcc
     2821 tggcctctct gacgtctagg acctccatgg gcattcttgt ggtcggagga gtggtgtgga
     2881 aggcagtggg ctggagactc atcgccctct cctttggact gtatggcctc ctgtacgtct
     2941 atgagcgact gacctggacc accaaagcca aagagagggc cttcaagcgc cagtttgtgg
     3001 aatacgccag tgagaagcta cagctcatca tcagttacac cggctctaac tgcagccacc
     3061 aagtccagca ggaattgtct gggacatttg ctcatctgtg ccagcaagtt gacatcaccc
     3121 gagataatct ggagcaggaa attgctgcca tgaacaagaa agtcgaggct ctggattcac
     3181 ttcagagcag agccaaactg ctcaggaata aagctggctg gttggacagc gaactcaaca
     3241 tgttcacaca ccagtacctg cagcccagca gaggatcctc tagaggtgaa caaaagttga
     3301 tttctgaaga agatttgaac ggtgaacaaa agctaatctc cgaggaagac ttgaacggtg
     3361 aacaaaaatt aatctcagaa gaagacttga acggatcctc tagaggtgaa caaaagttga
     3421 tttctgaaga agatttgaac ggtgaacaaa agctaatctc cgaggaagac ttgaacggtg
     3481 aacaaaaatt aatctcagaa gaagacttga acggatcctc tagaggtgaa caaaagttga
     3541 tttctgaaga agatttgaac ggtgaacaaa agctaatctc cgaggaagac ttgaacggtg
     3601 aacaaaaatt aatctcagaa gaagacttga acggatcctc tagaggtgaa caaaagttga
     3661 tttctgaaga agatttgaac ggtgaacaaa agctaatctc cgaggaagac ttgaacggtg
     3721 aacaaaaatt aatctcagaa gaagacttga acggatcctc tagaggtgaa caaaagttga
     3781 tttctgaaga agatttgaac ggtgaacaaa agctaatctc cgaggaagac ttgaacggtg
     3841 aacaaaaatt aatctcagaa gaagacttga acggatccga gctcggtacc aagcttgggc
     3901 ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc
     3961 atcattgagt ttaaacggtc tccagcttaa gtttaaaccg ctgatcagcc tcgactgtgc
     4021 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4081 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4141 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4201 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     4261 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4321 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4381 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4441 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4501 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4561 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4621 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     4681 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4741 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt
     4801 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4861 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4921 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4981 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5041 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5101 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5161 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5221 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5281 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5341 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5401 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5461 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5521 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5581 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5641 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5701 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5761 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5821 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5881 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5941 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6001 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6061 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6241 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6841 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7081 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7261 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7321 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7381 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7441 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7501 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7561 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7621 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7681 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7741 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7801 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7861 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7921 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7981 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8041 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8101 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8161 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8221 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8281 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8341 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8401 aaaagtgcca cctgacgtc
//
