LOCUS       V030465                 8280 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030465
VERSION     V030465
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             756..2960
                     /label="Rnasel(NM_011882)"
                     /note="Rnasel(NM_011882)"
                     /gene="Rnasel"
     CDS             2967..3032
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3056..3644
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3710..3758
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3960..4388
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4402..4731
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4798..5592
                     /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    5768..5889
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5938..5954)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5962..5978
                     /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(5986..6016)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6031..6052
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6340..6928)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7099..7959)
                     /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(7960..8064)
                     /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 ctcgaattcg ccaccatgga gaccccggat tataacacac
      781 ctcagggtgg aaccccatca gcgggaagtc agaggaccgt tgtcgaagat gattcttcgt
      841 tgatcaaagc tgttcagaag ggagatgttg tcagggtcca gcaattgtta gaaaaagggg
      901 ctgatgccaa tgcctgtgaa gacacctggg gctggacacc tttgcacaac gcagtgcaag
      961 ctggcagggt agacattgtg aacctcctgc ttagtcatgg tgctgaccct catcggagga
     1021 agaagaatgg ggccaccccc ttcatcattg ctgggatcca gggagatgtg aaactgctcg
     1081 agattctcct ctcttgtggt gcagacgtca atgagtgtga cgagaacgga ttcacggctt
     1141 tcatggaagc tgctgagcgt ggtaacgctg aagccttaag attccttttt gctaagggag
     1201 ccaatgtgaa tttgcgacga cagacaacga aggacaaaag gcgattgaag caaggaggcg
     1261 ccacagctct catgagcgct gctgagaagg gccacctgga agtcctgaga attctcctca
     1321 atgacatgaa ggcagaagtc gatgctcggg acaacatggg cagaaatgcc ctgatccgta
     1381 ctctgctgaa ctgggattgt gaaaatgtgg aggagattac ttcaatcctg attcagcacg
     1441 gggctgatgt taacgtgaga ggagaaagag ggaaaacacc cctcatcgca gcagtggaga
     1501 ggaagcacac aggcttggtg cagatgctcc tgagtcggga aggcataaac atagatgcca
     1561 gggataacga gggcaagaca gctctgctaa ttgctgttga taaacaactg aaggaaattg
     1621 tccagttgct tcttgaaaag ggagctgata agtgtgacga tcttgtttgg atagccagga
     1681 ggaatcatga ctatcacctt gtaaagcttc tcctccctta tgtagctaat cctgacaccg
     1741 accctcctgc tggagactgg tcgcctcaca gttcacgttg ggggacagcc ttgaaaagcc
     1801 tccacagtat gactcgaccc atgattggca aactcaagat cttcattcat gatgactata
     1861 aaattgctgg cacttccgaa ggggctgtct acctagggat ctatgacaat cgagaagtgg
     1921 ctgtgaaggt cttccgtgag aatagcccac gtggatgtaa ggaagtctct tgtctgcggg
     1981 actgcggtga ccacagtaac ttagtggctt tctatggaag agaggacgat aagggctgtt
     2041 tatatgtgtg tgtgtccctg tgtgagtgga cactggaaga gttcctgagg ttgcccagag
     2101 aggaacctgt ggagaacggg gaagataagt ttgcccacag catcctatta tctatatttg
     2161 agggtgttca aaaactacac ttgcatggat attcccatca ggacctgcaa ccacaaaaca
     2221 tcttaataga ttccaagaaa gctgtccggc tggcagattt tgatcagagc atccgatgga
     2281 tgggagagtc acagatggtc aggagagact tggaggatct tggacggctg gttctctacg
     2341 tggtaatgaa aggtgagatc ccctttgaga cactaaagac tcagaatgat gaagtgctgc
     2401 ttacaatgtc tccagatgag gagactaagg acctcattca ttgcctgttt tctcctggag
     2461 aaaatgtcaa gaactgcctg gtagacctgc ttggccatcc tttcttttgg acttgggaga
     2521 accgctatag aacactccgg aatgtgggaa atgaatctga catcaaagta cggaaatgta
     2581 aaagtgatct tctcagacta ctgcagcatc agacacttga gcctcccaga agctttgacc
     2641 agtggacatc taagatcgac aaaaatgtta tggatgaaat gaatcatttc tacgaaaaga
     2701 gaaaaaaaaa cccttatcag gatactgtag gtgatctgct gaagtttatt cggaatatag
     2761 gcgaacacat caatgaggaa aaaaagcggg gaatgaagga gatacttgga gatccttctc
     2821 gttattttca ggagacattt ccagatcttg taatctatat ctacaagaaa ctaaaggaaa
     2881 cagaatacag aaaacacttc ccacagcctc caccacgcct cagcgttcca gaggcagtgg
     2941 gaccaggggg catacagagc ggatccgact acaaagacca tgacggtgat tataaagatc
     3001 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     3061 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3121 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3181 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3241 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3301 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3361 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3421 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3481 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3541 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3601 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3661 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3721 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3781 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3841 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3901 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3961 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4021 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4081 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     4141 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4201 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4261 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4321 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4381 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4441 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4561 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4621 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4681 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4741 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4801 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4861 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4921 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4981 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5041 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5101 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5161 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5221 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5281 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5341 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5401 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5461 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5521 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5581 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5641 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5701 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5761 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5821 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5881 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5941 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6001 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6061 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6121 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6181 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6241 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6301 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6361 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6421 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6481 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6541 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6601 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6661 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6721 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6781 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6841 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6901 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6961 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7021 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7081 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7141 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7201 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7261 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7321 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7381 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7441 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7501 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7561 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7621 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7681 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7741 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7801 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7861 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7921 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7981 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8041 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8101 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     8161 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     8221 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
