LOCUS       V023978                 8321 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023978
VERSION     V023978
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             779..3001
                     /label="Mfn1(NM_024200)"
                     /note="Mfn1(NM_024200)"
                     /gene="Mfn1"
     CDS             3008..3073
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3097..3685
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3751..3799
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4001..4429
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4443..4772
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4839..5633
                     /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    5809..5930
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5979..5995)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6003..6019
                     /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(6027..6057)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6072..6093
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6381..6969)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7140..8000)
                     /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(8001..8105)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 ggcagaaacg gtatctccac tgaagcactt tgtgctggca aagaaagcca tcactgcaat
      841 cttcggccag ttactggagt ttgttactga gggctcacat tttgttgaag caacatacag
      901 gaatccagaa cttgatcgaa tagcatccga ggatgatctg gtggaaatac agggctacag
      961 aaacaagctt gctgtcattg gggaggtgct gtctcggaga catatgaagg tggcattttt
     1021 tggcaggaca agtagtggca agagctctgt catcaatgca atgctgtggg ataaagtcct
     1081 ccccagcggg attggtcaca caaccaactg cttcctgagt gtcgagggga ccgatggaga
     1141 taaagcctac cttatgaccg aagggtcaga tgaaaagaaa agtgtgaaga ctgttaatca
     1201 gctggcccat gccctccata tggataaaga cttgaaagct ggctgtcttg tgcatgtatt
     1261 ttggcccaaa gcaaaatgtg ccctcttgag agatgacctg gttttagtag acagcccagg
     1321 tacagatgtc accacagagc tggatatctg gattgataag ttttgccttg atgctgatgt
     1381 ctttgttttg gttgcaaact cggaatcaac actgatgaac acggagaaac attttttcca
     1441 taaggtgaat gagcggctct ccaagcccaa catcttcatt ctgaataacc gttgggatgc
     1501 ttctgcttcg gagccggagt acatggagga tgtgcgcaga cagcacatgg agagatgtct
     1561 tcacttcttg gtagaagagc tcaaggttgt aagtccgtcg gaagctcgga atcggatctt
     1621 ttttgtttca gccaaggaag ttctcaactc cagaaagcat aaagctcagg ggatgccaga
     1681 aggtggtggg gcacttgcag aaggatttca agcaagatta caggagtttc aaaattttga
     1741 acaaactttt gaggagtgta tctcgcagtc agcagtgaaa acaaagtttg aacagcacac
     1801 tatcagagct aaacagatac tagacactgt gaaaaacata ctggactcag taaacgtggc
     1861 agcagcagag aagagggttt attcaatgga agagagggaa gaccaaatcg atagactgga
     1921 ctttatccga aaccagatga accttttaac actggatgtt aagaagaaga tcaaggaggt
     1981 cacggaggag gtggcaaaca aggtttcttg tgcaatgaca gatgaaattt gtcgactatc
     2041 tgttttggtt gatgagtttt gttctgagtt tcatcctacc cccagtgtac tgaaagtgta
     2101 taagagtgag ttaaataagc acatagaaga tggcatggga agaaatttgg ctgatcggtg
     2161 taccaatgaa gtcaatgcct ccattcttca atctcagcaa gaaatcatcg aaaacttgaa
     2221 gccactactt ccagctggta tacagaataa acttcataca ttaatccctt gcaaaaagtt
     2281 tgacctcagc tatgatctca attgccacaa gctgtgttcg gattttcaag aggacattgt
     2341 gtttcggttt tccctgggct ggtcttccct tgtacatcga ttcctgggtt ccacaaatgc
     2401 acagagggtg ctgctcgggc tgtcagagcc catctttcag gtccctagat ctttagcttc
     2461 aactcctact gctccttcta acccagcagc cccggataat gcagcccagg aggagctcat
     2521 gatcaccctg atcacaggat tggcgtccct cacgtcgaga acctccatgg gcatcatcgt
     2581 tgttgggggc gtgatttgga aaacagtggg ctggaaacta atctctgtca ccttaagtat
     2641 gtacggagct ctgtaccttt atgagaggct gacgtggacg acccgtgcga aagagagagc
     2701 gtttaagcag cagtttgtaa actatgcaac cgagaagctg cagatgattg tgagcttcac
     2761 cagtgcaaac tgcagccacc aagtacagca agaaatggcc actacttttg ctcgactgtg
     2821 ccaacaagtt gatgttactc agaaacatct ggaagaggaa attgcaagat tatccaaaga
     2881 gatagaccaa ctggagaaaa tacagaacaa ctcaaagctc ttaagaaata aagctgttca
     2941 acttgaaagt gagctggaga atttttcgaa gcagtttcta cacccgagca gtggagaatc
     3001 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     3061 tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt
     3121 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3181 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3241 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3301 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3361 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3421 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3481 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     3541 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3601 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3661 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg
     3721 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     3781 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     3841 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     3901 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     3961 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4021 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4081 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4141 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4201 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4261 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4321 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     4381 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4441 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4501 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4561 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4621 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4681 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4741 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4801 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4861 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4921 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4981 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5041 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5101 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5161 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5221 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5281 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5341 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5401 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5461 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5521 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5581 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5641 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5701 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5761 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5821 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5881 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5941 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6001 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6061 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6121 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6181 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6301 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6541 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     6601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6781 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7021 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7081 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7141 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7201 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7261 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7321 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7381 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7441 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7501 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7561 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7621 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7681 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7741 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7801 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7861 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7921 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7981 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8101 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     8161 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     8221 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     8281 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
