LOCUS       V027685                 8467 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027685
VERSION     V027685
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..3123
                     /label="VCP(NM_007126)"
                     /note="VCP(NM_007126)"
                     /gene="VCP"
     CDS             3130..3219
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3243..3831
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3897..3945
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4147..4575
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4589..4918
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4985..5779
                     /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    5955..6076
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6125..6141)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6149..6165
                     /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(6173..6203)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6218..6239
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6527..7115)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7286..8146)
                     /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(8147..8251)
                     /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 ccaccatggc ttctggagcc
      721 gattcaaaag gtgatgacct atcaacagcc attctcaaac agaagaaccg tcccaatcgg
      781 ttaattgttg atgaagccat caatgaggac aacagtgtgg tgtccttgtc ccagcccaag
      841 atggatgaat tgcagttgtt ccgaggtgac acagtgttgc tgaaaggaaa gaagagacga
      901 gaagctgttt gcatcgtcct ttctgatgat acttgttctg atgagaagat tcggatgaat
      961 agagttgttc ggaataacct tcgtgtacgc ctaggggatg tcatcagcat ccagccatgc
     1021 cctgatgtga agtacggcaa acgtatccat gtgctgccca ttgatgacac agtggaaggc
     1081 attactggta atctcttcga ggtatacctt aagccgtact tcctggaagc gtatcgaccc
     1141 atccggaaag gagacatttt tcttgtccgt ggtgggatgc gtgctgtgga gttcaaagtg
     1201 gtggaaacag atcctagccc ttattgcatt gttgctccag acacagtgat ccactgcgaa
     1261 ggggagccta tcaaacgaga ggatgaggaa gagtccttga atgaagtagg gtatgatgac
     1321 attggtggct gcaggaagca gctagctcag ataaaggaga tggtggaact gcccctgaga
     1381 catcctgccc tctttaaggc aattggtgtg aagcctccta gaggaatcct gctttacgga
     1441 cctcctggaa caggaaagac cctgattgct cgagctgtag caaatgagac tggagccttc
     1501 ttcttcttga tcaatggtcc tgagatcatg agcaaattgg ctggtgagtc tgagagcaac
     1561 cttcgtaaag cctttgagga ggctgagaag aatgctcctg ccatcatctt cattgatgag
     1621 ctagatgcca tcgctcccaa aagagagaaa actcatggcg aggtggagcg gcgcattgta
     1681 tcacagttgt tgaccctcat ggatggccta aagcagaggg cacatgtgat tgttatggca
     1741 gcaaccaaca gacccaacag cattgaccca gctctacggc gatttggtcg ctttgacagg
     1801 gaggtagata ttggaattcc tgatgctaca ggacgcttag agattcttca gatccatacc
     1861 aagaacatga agctggcaga tgatgtggac ctggaacagg tagccaatga gactcacggg
     1921 catgtgggtg ctgacttagc agccctgtgc tcagaggctg ctctgcaagc catccgcaag
     1981 aagatggatc tcattgacct agaggatgag accattgatg ccgaggtcat gaactctcta
     2041 gcagttacta tggatgactt ccggtgggcc ttgagccaga gtaacccatc agcactgcgg
     2101 gaaaccgtgg tagaggtgcc acaggtaacc tgggaagaca tcgggggcct agaggatgtc
     2161 aaacgtgagc tacaggagct ggtccagtat cctgtggagc acccagacaa attcctgaag
     2221 tttggcatga caccttccaa gggagttctg ttctatggac ctcctggctg tgggaaaact
     2281 ttgttggcca aagccattgc taatgaatgc caggccaact tcatctccat caagggtcct
     2341 gagctgctca ccatgtggtt tggggagtct gaggccaatg tcagagaaat ctttgacaag
     2401 gcccgccaag ctgccccctg tgtgctattc tttgatgagc tggattcgat tgccaaggct
     2461 cgtggaggta acattggaga tggtggtggg gctgctgacc gagtcatcaa ccagatcctg
     2521 acagaaatgg atggcatgtc cacaaaaaaa aatgtgttca tcattggcgc taccaaccgg
     2581 cctgacatca ttgatcctgc catcctcaga cctggccgtc ttgatcagct catctacatc
     2641 ccacttcctg atgagaagtc ccgtgttgcc atcctcaagg ctaacctgcg caagtcccca
     2701 gttgccaagg atgtggactt ggagttcctg gctaaaatga ctaatggctt ctctggagct
     2761 gacctgacag agatttgcca gcgtgcttgc aagctggcca tccgtgaatc catcgagagt
     2821 gagattaggc gagaacgaga gaggcagaca aacccatcag ccatggaggt agaagaggat
     2881 gatccagtgc ctgagatccg tcgagatcac tttgaagaag ccatgcgctt tgcgcgccgt
     2941 tctgtcagtg acaatgacat tcggaagtat gagatgtttg cccagaccct tcagcagagt
     3001 cggggctttg gcagcttcag attcccttca gggaaccagg gtggagctgg ccccagtcag
     3061 ggcagtggag gcggcacagg tggcagtgta tacacagaag acaatgatga tgacctgtat
     3121 ggcctcgagg aacaaaaact aatatcagag gaagatttgg aacagaaact aatcagtgaa
     3181 gaagatttag agcaaaagtt aatttccgag gaggacttat agtgagggcc cgatatctcg
     3241 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3301 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3361 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3421 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3481 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3541 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3601 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     3661 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3721 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3781 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg
     3841 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     3901 aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc
     3961 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4021 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4081 ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt
     4141 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4201 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4261 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc
     4321 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4381 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4441 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4501 atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4561 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4621 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4681 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4741 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4801 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4861 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4921 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     4981 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5041 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5101 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5161 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5221 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5281 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5341 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5401 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5461 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5521 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5581 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5641 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5701 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5761 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     5821 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5881 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5941 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6001 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6061 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6121 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6181 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6241 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6301 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6361 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6421 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6481 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6541 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6601 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6661 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6721 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6781 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6841 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6901 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6961 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7021 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7081 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7141 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7201 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7261 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7321 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7381 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7441 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7501 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7561 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7621 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7681 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7741 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7801 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7861 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7921 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7981 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8041 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8101 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8161 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8221 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8281 tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga
     8341 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     8401 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     8461 gcttagg
//
