LOCUS       V043346                 8317 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043346
VERSION     V043346
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"
     CDS             706..2976
                     /label="Mfn2(NM_133201)"
                     /note="Mfn2(NM_133201)"
                     /gene="Mfn2"
     CDS             2983..3009
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3013..3039
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3043..3069
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3093..3681
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3747..3795
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3997..4425
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4439..4768
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4835..5629
                     /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    5805..5926
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5975..5991)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5999..6015
                     /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(6023..6053)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6068..6089
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6377..6965)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7136..7996)
                     /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(7997..8101)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgtc cctgctcttt
      721 tctcgatgca actccatcgt caccgtcaag aaggataagc gacacatggc tgaagtgaat
      781 gcttcccctc tcaagcactt tgtcactgcc aagaaaaaga tcaatggaat ctttgagcag
      841 ctgggggcct acatccaaga gagcgccagc ttccttgaag acacccacag gaacacagaa
      901 ctggacccgg ttaccacgga agagcaggtc ctggacgtca aagggtacct gtccaaggtc
      961 aggggtatca gcgaagtgct ggccaggcgg cacatgaagg tggctttttt tggccggacg
     1021 agcaatggga agagcaccgt gatcaatgcc atgctctggg acaaagttct gccatctggg
     1081 attggtcata ccaccaattg cttcctgcgg gttgggggca cagatggcca tgaggccttc
     1141 ctcctcacag agggctcaga agagaagaag agtgtcaaga ctgtgaacca actggcccat
     1201 gccctccatc aggacgagca gttgcatgca ggcagcatgg tgagtgtgat gtggcccaac
     1261 tccaagtgtc cgctcctgaa ggatgacctc gtgctgatgg acagccctgg gatcgatgtt
     1321 accacggagc tggacagctg gattgataag ttttgcctgg atgctgatgt gtttgtgctg
     1381 gtggccaact cagagtccac gctgatgcag acggagaagc agttcttcca caaagtgagt
     1441 gaacgtctct cccggcccaa catcttcatc ctgaacaacc gctgggatgc gtctgcctcg
     1501 gagcctgagt acatggagga ggtgcggcgg cagcacatgg agcgctgcac cagctttctg
     1561 gtggatgagc tgggcgtggt ggatcgagct caggctgggg accggatctt cttcgtgtct
     1621 gccaaggagg ttctcagcgc cagggtccag aaagcccagg gcatgccaga aggaggcggc
     1681 gctctcgcag aaggttttca agtgaggatg tttgagtttc agaatttcga gaggcagttt
     1741 gaggagtgca tttcccagtc tgcagtaaag accaaatttg agcagcacac agtccgggcc
     1801 aagcagattg cagaggccgt tcgtctcatc atggattccc tgcacatcgc agctcaggag
     1861 cagcgggttt attgcctaga aatgcgggaa gagcggcaag accggctgag gtttattgac
     1921 aagcagctgg agctcctggc tcaagactac aagctgcgaa ttaagcagat tacggaggaa
     1981 gtggaaaggc aggtgtccac agccatggct gaagagatca ggcgcctctc tgtgctagtt
     2041 gacgagtacc agatggactt ccacccatcc ccagttgtcc tcaaggttta taagaacgag
     2101 ctgcaccgcc atatagagga aggtctgggc cggaacctgt ctgaccgctg ctccactgcc
     2161 attgccagtt cactgcagac tatgcagcag gacatgatag acggcttgaa gccccttctt
     2221 cctgtatcta tgcggaatca gatagacatg ctggtccctc gacagtgttt ctccctcagc
     2281 tatgacctga attgtgacaa gctgtgtgct gactttcagg aggacatcga gttccacttc
     2341 tcccttggat ggactatgct agtgaacagg ttcctgggcc ccaagaatag ccgccgggcc
     2401 ttgctaggct acagtgatca ggttcagcgt cctctccctc tgacacctgc caaccccagc
     2461 atgcccccct tgccacagag ctccctcacc caggaggagc tcatggtctc catggttact
     2521 ggcctggcct ctctgacgtc taggacctcc atgggcattc ttgtggtcgg aggagtggtg
     2581 tggaaggcag tgggctggag actcatcgcc ctctcctttg gactgtatgg cctcctgtac
     2641 gtctatgagc gactgacctg gaccaccaaa gccaaagaga gggccttcaa gcgccagttt
     2701 gtggaatacg ccagtgagaa gctacagctc atcatcagtt acaccggctc taactgcagc
     2761 caccaagtcc agcaggaatt gtctgggaca tttgctcatc tgtgccagca agttgacatc
     2821 acccgagata atctggagca ggaaattgct gccatgaaca agaaagtcga ggctctggat
     2881 tcacttcaga gcagagccaa actgctcagg aataaagctg gctggttgga cagcgaactc
     2941 aacatgttca cacaccagta cctgcagccc agcagactcg agtacccata cgacgtacct
     3001 gattatgcaa cttatcctta tgatgtacct gattatgcta gctatcctta tgatgtccca
     3061 gactacgcat agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg
     3121 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3181 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3241 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3301 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3361 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3421 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3481 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3541 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3601 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3661 ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac
     3721 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     3781 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     3841 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     3901 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     3961 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4021 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4081 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4141 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4201 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4261 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4321 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     4381 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4441 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4501 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4561 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4621 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4681 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4741 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4801 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4861 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4921 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4981 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5041 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5101 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5161 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5221 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5281 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5341 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5401 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5461 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5521 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5581 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5641 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5701 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5761 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5821 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5881 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5941 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6001 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6061 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6121 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6181 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6241 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6301 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6361 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6421 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6481 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6541 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     6601 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6661 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6721 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6781 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6841 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6901 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6961 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7021 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7081 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7141 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7201 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7261 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7321 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7381 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7441 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7501 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7561 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7621 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7681 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7741 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7801 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7861 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7921 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7981 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8041 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8101 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     8161 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     8221 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     8281 aggcttgacc gacaattgca tgaagaatct gcttagg
//
