LOCUS       pcDNA3.1-MFN2(human)    7747 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023070
VERSION     V023070
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3234
                     /label="MFN2(NM_014874)"
                     /note="MFN2(NM_014874)"
                     /gene="MFN2"
     CDS             3244..3309
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3347..3571
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3617..4045
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4059..4388
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4455..5249
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5423..5544
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5593..5609)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5617..5633
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5641..5671)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5686..5707
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5995..6580)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6751..7611)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7612..7716)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgtccc tgctcttctc tcgatgcaac tctatcgtca cagtcaagaa aaataagaga
     1021 cacatggctg aggtgaatgc atccccactt aagcactttg tcactgccaa gaagaagatc
     1081 aatggcattt ttgagcagct gggggcctac atccaggaga gcgccacctt ccttgaagac
     1141 acgtacagga atgcagaact ggaccccgtt accacagaag aacaggttct ggacgtcaaa
     1201 ggttacctat ccaaagtgag aggcatcagt gaggtgctgg ctcggaggca catgaaagtg
     1261 gctttttttg gccggacgag caatgggaag agcaccgtga tcaatgccat gctctgggac
     1321 aaagttctgc cctctgggat tggccacacc accaattgct tcctgcgggt agagggcaca
     1381 gatggccatg aggcctttct ccttaccgag ggctcagagg aaaagaggag tgccaagact
     1441 gtgaaccagc tggcccatgc cctccaccag gacaagcagc tccatgccgg cagcctagtg
     1501 agtgtgatgt ggcccaactc taagtgccca cttctgaagg atgacctcgt tttgatggac
     1561 agccctggta ttgatgtcac cacagagctg gacagctgga ttgacaagtt ttgtctggat
     1621 gctgatgtgt ttgtgctggt ggccaactca gagtccaccc tgatgcagac ggaaaagcac
     1681 ttcttccaca aggtgagtga gcgtctctcc cggccaaaca tcttcatcct gaacaaccgc
     1741 tgggatgcat ctgcctcaga gcccgagtac atggaggagg tgcggcggca gcacatggag
     1801 cgttgtacca gcttcctggt ggatgagctg ggcgtggtgg atcgatccca ggccggggac
     1861 cgcatcttct ttgtgtctgc taaggaggtg ctcaacgcca ggattcagaa agcccagggc
     1921 atgcctgaag gagggggcgc tctcgcagaa ggctttcaag tgaggatgtt tgagtttcag
     1981 aattttgaga ggagatttga ggagtgcatc tcccagtctg cagtgaagac caagtttgag
     2041 cagcacacgg tccgggccaa gcagattgca gaggcggttc gactcatcat ggactccctg
     2101 cacatggcgg ctcgggagca gcaggtttac tgcgaggaaa tgcgtgaaga gcggcaagac
     2161 cgactgaaat ttattgacaa acagctggag ctcttggctc aagactataa gctgcgaatt
     2221 aagcagatta cggaggaagt ggagaggcag gtgtcgactg caatggccga ggagatcagg
     2281 cgcctctctg tactggtgga cgattaccag atggacttcc acccttctcc agtagtcctc
     2341 aaggtttata agaatgagct gcaccgccac atagaggaag gactgggtcg aaacatgtct
     2401 gaccgctgct ccacggccat caccaactcc ctgcagacca tgcagcagga catgatagat
     2461 ggcttgaaac ccctccttcc tgtgtctgtg cggagtcaga tagacatgct ggtcccacgc
     2521 cagtgcttct ccctcaacta tgacctaaac tgtgacaagc tgtgtgctga cttccaggaa
     2581 gacattgagt tccatttctc tctcggatgg accatgctgg tgaataggtt cctgggcccc
     2641 aagaacagcc gtcgggcctt gatgggctac aatgaccagg tccagcgtcc catccctctg
     2701 acgccagcca accccagcat gcccccactg ccacagggct cgctcaccca ggaggagttc
     2761 atggtttcca tggttaccgg cctggcctcc ttgacatcca ggacctccat gggcattctt
     2821 gttgttggag gagtggtgtg gaaggcagtg ggctggcggc tcattgccct ctcctttggg
     2881 ctctatggcc tcctctacgt ctatgagcgt ctgacctgga ccaccaaggc caaggagagg
     2941 gccttcaagc gccagtttgt ggagcatgcc agcgagaagc tgcagcttgt catcagctac
     3001 actggctcca actgcagcca ccaagtccag caggaactgt ctgggacctt tgctcatctg
     3061 tgtcagcaag ttgacgtcac ccgggagaac ctggagcagg aaattgccgc catgaacaag
     3121 aaaattgagg ttcttgactc acttcagagc aaagcaaagc tgctcaggaa taaagccggt
     3181 tggttggaca gtgagctcaa catgttcaca caccagtacc tgcagcccag cagatagctc
     3241 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3301 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3361 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3421 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3481 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3541 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3601 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3661 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3721 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3781 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3841 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3901 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3961 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4021 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4081 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4141 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4261 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4321 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4381 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4441 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4501 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4561 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4621 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4681 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4741 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4801 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4861 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4921 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4981 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5041 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5101 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5161 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5221 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5281 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5341 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5401 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5461 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5521 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5581 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5641 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5701 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5821 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5881 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5941 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6181 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6421 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7741 tgacgtc
//
