LOCUS       V019891                 8738 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019891
VERSION     V019891
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             650..2641
                     /label="LMNA(NM_170707)"
                     /note="LMNA(NM_170707)"
                     /gene="LMNA"
     CDS             2669..3385
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced YFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             3425..3490
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3514..4102
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4168..4216
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4418..4846
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4860..5189
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5256..6050
                     /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    6226..6347
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6396..6412)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6420..6436
                     /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(6444..6474)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6489..6510
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6798..7386)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7557..8417)
                     /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(8418..8522)
                     /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 gctgccacca tggagacccc
      661 gtcccagcgg cgcgccaccc gcagcggggc gcaggccagc tccactccgc tgtcgcccac
      721 ccgcatcacc cggctgcagg agaaggagga cctgcaggag ctcaatgatc gcttggcggt
      781 ctacatcgac cgtgtgcgct cgctggaaac ggagaacgca gggctgcgcc ttcgcatcac
      841 cgagtctgaa gaggtggtca gccgcgaggt gtccggcatc aaggccgcct acgaggccga
      901 gctcggggat gcccgcaaga cccttgactc agtagccaag gagcgcgccc gcctgcagct
      961 ggagctgagc aaagtgcgtg aggagtttaa ggagctgaaa gcgcgcaata ccaagaagga
     1021 gggtgacctg atagctgctc aggctcggct gaaggacctg gaggctctgc tgaactccaa
     1081 ggaggccgca ctgagcactg ctctcagtga gaagcgcacg ctggagggcg agctgcatga
     1141 tctgcggggc caggtggcca agcttgaggc agccctaggt gaggccaaga agcaacttca
     1201 ggatgagatg ctgcggcggg tggatgctga gaacaggctg cagaccatga aggaggaact
     1261 ggacttccag aagaacatct acagtgagga gctgcgtgag accaagcgcc gtcatgagac
     1321 ccgactggtg gagattgaca atgggaagca gcgtgagttt gagagccggc tggcggatgc
     1381 gctgcaggaa ctgcgggccc agcatgagga ccaggtggag cagtataaga aggagctgga
     1441 gaagacttat tctgccaagc tggacaatgc caggcagtct gctgagagga acagcaacct
     1501 ggtgggggct gcccacgagg agctgcagca gtcgcgcatc cgcatcgaca gcctctctgc
     1561 ccagctcagc cagctccaga agcagctggc agccaaggag gcgaagcttc gagacctgga
     1621 ggactcactg gcccgtgagc gggacaccag ccggcggctg ctggcggaaa aggagcggga
     1681 gatggccgag atgcgggcaa ggatgcagca gcagctggac gagtaccagg agcttctgga
     1741 catcaagctg gccctggaca tggagatcca cgcctaccgc aagctcttgg agggcgagga
     1801 ggagaggcta cgcctgtccc ccagccctac ctcgcagcgc agccgtggcc gtgcttcctc
     1861 tcactcatcc cagacacagg gtgggggcag cgtcaccaaa aagcgcaaac tggagtccac
     1921 tgagagccgc agcagcttct cacagcacgc acgcactagc gggcgcgtgg ccgtggagga
     1981 ggtggatgag gagggcaagt ttgtccggct gcgcaacaag tccaatgagg accagtccat
     2041 gggcaattgg cagatcaagc gccagaatgg agatgatccc ttgctgactt accggttccc
     2101 accaaagttc accctgaagg ctgggcaggt ggtgacgatc tgggctgcag gagctggggc
     2161 cacccacagc ccccctaccg acctggtgtg gaaggcacag aacacctggg gctgcgggaa
     2221 cagcctgcgt acggctctca tcaactccac tggggaagaa gtggccatgc gcaagctggt
     2281 gcgctcagtg actgtggttg aggacgacga ggatgaggat ggagatgacc tgctccatca
     2341 ccaccacggc tcccactgca gcagctcggg ggaccccgct gagtacaacc tgcgctcgcg
     2401 caccgtgctg tgcgggacct gcgggcagcc tgccgacaag gcatctgcca gcggctcagg
     2461 agcccaggtg ggcggaccca tctcctctgg ctcttctgcc tccagtgtca cggtcactcg
     2521 cagctaccgc agtgtggggg gcagtggggg tggcagcttc ggggacaatc tggtcacccg
     2581 ctcctacctc ctgggcaact ccagcccccg aacccagagc ccccagaact gcagcatcat
     2641 gggaggaggg ggaagcggat ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     2701 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2761 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2821 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgaagtg
     2881 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2941 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3001 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3061 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     3121 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     3181 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3241 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     3301 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3361 tctcggcatg gacgagctgt acaaggaatt ctgcagatat ccagcacagt ggcggccgct
     3421 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     3481 cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     3541 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3601 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3661 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3721 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3781 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3841 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3901 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     3961 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4021 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4081 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     4141 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     4201 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4261 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4321 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4381 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4441 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4501 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4561 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4621 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4681 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4741 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     4801 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4861 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4921 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4981 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5041 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5101 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5161 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5221 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5281 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5341 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5401 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5461 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5521 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5581 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5641 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5701 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5761 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5821 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5881 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5941 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6001 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6061 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6121 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6181 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6241 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6301 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6361 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6421 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6481 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6541 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6601 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6661 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6721 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6781 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6841 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6901 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6961 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     7021 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7081 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7141 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7201 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7261 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7321 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7381 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7441 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7501 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7561 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7621 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7681 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7741 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7801 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7861 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7921 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7981 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8041 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8101 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8161 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8221 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8281 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8341 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8401 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8461 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8521 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     8581 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     8641 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     8701 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
