LOCUS       V019150                 8750 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019150
VERSION     V019150
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..3430
                     /label="OGA(NM_012215)"
                     /note="OGA(NM_012215)"
                     /gene="OGA"
     CDS             3437..3502
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3526..4114
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4180..4228
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4430..4858
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4872..5201
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5268..6062
                     /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    6238..6359
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6408..6424)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6432..6448
                     /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(6456..6486)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6501..6522
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6810..7398)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7569..8429)
                     /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(8430..8534)
                     /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 ccatggtgca gaaggagagt caagcgacgt tggaggagcg
      721 ggagagcgag ctcagctcca accctgccgc ctctgcgggg gcatcgctgg agccgccggc
      781 agctccggca cccggagaag acaaccccgc cggggctggg ggagcggcgg tggccggggc
      841 tgcaggaggg gctcggcggt tcctctgcgg tgtggtggaa ggattttatg gaagaccttg
      901 ggttatggaa cagagaaaag aactctttag aaggctccag aaatgggaat taaatacata
      961 cttgtatgcc ccaaaagatg actacaaaca taggatgttt tggcgagaga tgtattcagt
     1021 ggaggaagct gagcaactta tgactctcat ctctgctgca cgagaatatg agatagagtt
     1081 catctatgcg atctcacctg gattggatat cactttttct aaccccaagg aagtatccac
     1141 attgaaacgt aaattggacc aggtttctca gtttgggtgc agatcatttg ctttgctttt
     1201 tgatgatata gaccataata tgtgtgcagc agacaaagag gtattcagtt cttttgctca
     1261 tgcccaagtc tccatcacaa atgaaatcta tcagtaccta ggagagccag aaactttcct
     1321 cttctgtccc acagaatact gtggcacttt ctgttatcca aatgtgtctc agtctccata
     1381 tttaaggact gtgggtgaaa agcttctacc tggaattgaa gtgctttgga caggtcccaa
     1441 agttgtttct aaagaaattc cagtagagtc catcgaagag gtttctaaga ttattaagag
     1501 agctccagta atctgggata acattcatgc taatgattat gatcagaaga gactgtttct
     1561 gggcccgtac aaaggaagat ccacagaact catcccacgg ttaaaaggag tcctcactaa
     1621 tccaaattgt gaatttgaag ccaactacgt tgctatccac acccttgcca cctggtacaa
     1681 atcaaacatg aatggagtga gaaaagatgt agtgatgact gacagtgaag atagtactgt
     1741 gtccatccag ataaaattag aaaatgaagg cagtgatgaa gatattgaaa ctgatgtact
     1801 ctatagtcca cagatggctc taaagctagc attaacagaa tggttgcaag agtttggtgt
     1861 gcctcatcaa tacagcagta ggcaagttgc acacagtgga gctaaagcaa gtgtagttga
     1921 tgggactcct ttagttgcag caccctcttt aaatgccaca accgtagtaa caacagttta
     1981 tcaggagccc attatgagcc agggagcagc cttgagtggt gagcctacta ctctgaccaa
     2041 ggaagaagaa aagaaacagc ctgatgaaga acccatggac atggtggtgg aaaaacaaga
     2101 agaaacggac cacaagaatg acaatcaaat actgagtgaa attgttgaag cgaaaatggc
     2161 agaggaattg aaaccaatgg acactgataa agagagcata gctgaatcaa aatccccaga
     2221 gatgtccatg caagaagatt gtattagtga cattgccccc atgcaaactg atgaacagac
     2281 aaacaaggag cagtttgtgc caggtccaaa tgaaaagcct ttgtacactg cggaaccagt
     2341 gaccctggag gatttgcagt tacttgctga tctattctac cttccttacg agcatggacc
     2401 caaaggagca cagatgttac gggaatttca atggcttcga gcaaatagta gtgttgtcag
     2461 tgtcaattgc aaaggaaaag actctgaaaa aattgaagaa tggcggtcac gagcagccaa
     2521 gtttgaagag atgtgtggac tagtgatggg aatgttcact cggctctcca attgtgccaa
     2581 caggacaatt ctttatgaca tgtactccta tgtttgggat atcaagagta taatgtctat
     2641 ggtgaagtct tttgtacagt ggttagggtg tcgtagtcat tcttcagcac aattcttaat
     2701 tggagaccaa gaaccctggg cctttagagg tggtctagca ggagagttcc agcgtttgct
     2761 gccaattgat ggggcaaatg atctcttttt tcagccacct ccactgactc ctacctccaa
     2821 agtttatact atcagacctt attttcctaa ggatgaggca tccgtgtaca agatttgcag
     2881 agaaatgtat gacgatggag tgggtttacc ctttcaaagt cagcctgatc ttattggaga
     2941 caagttagta ggagggctgc tttccctcag cctggattac tgctttgtcc tagaagatga
     3001 agatggcata tgtggttatg ccttgggcac tgtagatgtg acccccttta ttaaaaaatg
     3061 taaaatttcc tggatcccct tcatgcagga gaagtatacc aagccaaatg gtgacaagga
     3121 actctctgag gctgagaaaa taatgttgag tttccatgaa gaacaggaag tactgccaga
     3181 aactttcctt gctaatttcc cttctctgat aaagatggac attcacaaaa aagtaactga
     3241 cccaagtgtg gccaaaagca tgatggcttg cctcctgtct tcactgaagg ctaatggctc
     3301 ccggggagct ttctgtgaag tgagaccaga tgataaaaga attctggaat tttacagcaa
     3361 gttaggatgt tttgaaattg caaaaatgga aggatttcca aaggatgtgg ttatacttgg
     3421 tcggagcctg ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     3481 ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca acctctggat
     3541 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3601 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3661 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3721 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3781 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3841 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3901 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     3961 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4021 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4081 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     4141 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4201 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4261 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4321 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4381 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     4441 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4501 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4561 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     4621 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4681 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4741 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4801 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4861 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4921 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4981 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5041 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5101 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     5161 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     5221 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5281 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5341 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5401 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5461 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5521 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5581 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5641 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5701 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5761 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5821 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5881 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5941 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6001 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6061 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6121 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6181 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     6241 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6301 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6361 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     6421 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6481 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6541 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6601 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6661 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6721 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6781 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6841 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6901 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6961 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     7021 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7081 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7141 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7201 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7261 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7321 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7381 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7441 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7501 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7561 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7621 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7681 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7741 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7801 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7861 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7921 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7981 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8041 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8101 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8161 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8221 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8281 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8341 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8401 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8461 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8521 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     8581 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     8641 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     8701 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
