LOCUS       V023685                 7793 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023685
VERSION     V023685
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             683..2440
                     /label="LMNB1(NM_005573)"
                     /note="LMNB1(NM_005573)"
                     /gene="LMNB1"
     CDS             2480..2545
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2569..3157
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3223..3271
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3473..3901
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3915..4244
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4311..5105
                     /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    5281..5402
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5451..5467)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5475..5491
                     /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(5499..5529)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5544..5565
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5853..6441)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6612..7472)
                     /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(7473..7577)
                     /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 ggatccgcca ccatggcgac tgcgaccccc gtgccgccgc ggatgggcag
      721 ccgcgctggc ggccccacca cgccgctgag ccccacgcgc ctgtcgcggc tccaggagaa
      781 ggaggagctg cgcgagctca atgaccggct ggcggtgtac atcgacaagg tgcgcagcct
      841 ggagacggag aacagcgcgc tgcagctgca ggtgacggag cgcgaggagg tgcgcggccg
      901 tgagctcacc ggcctcaagg cgctctacga gaccgagctg gccgacgcgc gacgcgcgct
      961 cgacgacacg gcccgcgagc gcgccaagct gcagatcgag ctgggcaagt gcaaggcgga
     1021 acacgaccag ctgctcctca actatgctaa gaaggaatct gatcttaatg gcgcccagat
     1081 caagcttcga gaatatgaag cagcactgaa ttcgaaagat gcagctcttg ctactgcact
     1141 tggtgacaaa aaaagtttag agggagattt ggaggatctg aaggatcaga ttgcccagtt
     1201 ggaagcctcc ttagctgcag ccaaaaaaca gttagcagat gaaactttac ttaaagtaga
     1261 tttggagaat cgttgtcaga gccttactga ggacttggag tttcgcaaaa gcatgtatga
     1321 agaggagatt aacgagacca gaaggaagca tgaaacgcgc ttggtagagg tggattctgg
     1381 gcgtcaaatt gagtatgagt acaagctggc gcaagccctt catgagatga gagagcaaca
     1441 tgatgcccaa gtgaggctgt ataaggagga gctggagcag acttaccatg ccaaacttga
     1501 gaatgccaga ctgtcatcag agatgaatac ttctactgtc aacagtgcca gggaagaact
     1561 gatggaaagc cgcatgagaa ttgagagcct ttcatcccag ctttctaatc tacagaaaga
     1621 gtctagagca tgtttggaaa ggattcaaga attagaggac ttgcttgcta aagaaaaaga
     1681 caactctcgt cgcatgctga cagacaaaga gagagagatg gcggaaataa gggatcaaat
     1741 gcagcaacag ctgaatgact atgaacagct tcttgatgta aagttagccc tggacatgga
     1801 aatcagtgct tacaggaaac tcttagaagg cgaagaagag aggttgaagc tgtctccaag
     1861 cccttcttcc cgtgtgacag tatcccgagc atcctcaagt cgtagtgtac gtacaactag
     1921 aggaaagcgg aagagggttg atgtggaaga atcagaggcg agtagtagtg ttagcatctc
     1981 tcattccgcc tcagccactg gaaatgtttg catcgaagaa attgatgttg atgggaaatt
     2041 tatccgcttg aagaacactt ctgaacagga tcaaccaatg ggaggctggg agatgatcag
     2101 aaaaattgga gacacatcag tcagttataa atatacctca agatatgtgc tgaaggcagg
     2161 ccagactgtt acaatttggg ctgcaaacgc tggtgtcaca gccagccccc caactgacct
     2221 catctggaag aaccagaact cgtggggcac tggcgaagat gtgaaggtta tattgaaaaa
     2281 ttctcaggga gaggaggttg ctcaaagaag tacagtcttt aaaacaacca tacctgaaga
     2341 agaggaggag gaggaagaag cagctggagt ggttgttgag gaagaacttt tccaccagca
     2401 gggaacccca agagcatcca atagaagctg tgcaattatg gaattctgca gatatccagc
     2461 acagtggcgg ccgctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     2521 cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa tcaacctctg
     2581 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2641 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2701 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2761 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2821 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2881 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2941 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3001 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3061 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3121 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg
     3181 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     3241 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     3301 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     3361 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     3421 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     3481 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3541 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3601 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     3661 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3721 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3781 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3841 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3901 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3961 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4021 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4081 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4141 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4201 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4261 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4321 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4381 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4441 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4501 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4561 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4621 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4681 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4741 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4801 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4861 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4921 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4981 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5041 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5101 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     5161 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5221 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5281 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5341 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5401 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5461 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5521 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5581 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5641 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5701 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5761 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5821 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5881 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5941 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6001 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     6061 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6121 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6181 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6241 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6301 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6361 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6421 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6481 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6541 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6601 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6661 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6721 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6781 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6841 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6901 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6961 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7021 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7081 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7141 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7201 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7261 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7321 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7381 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7441 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7501 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7561 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     7621 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     7681 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     7741 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
