LOCUS       V029032                 7300 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029032
VERSION     V029032
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..2820
                     /label="PTGS2(NM_000963)"
                     /note="PTGS2(NM_000963)"
                     /gene="PTGS2"
     polyA_signal    2900..3124
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3170..3598
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3612..3941
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4008..4802
                     /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    4976..5097
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5146..5162)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5170..5186
                     /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(5194..5224)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5239..5260
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5548..6133)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6304..7164)
                     /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(7165..7269)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat gctcgcccgc
     1021 gccctgctgc tgtgcgcggt cctggcgctc agccatacag caaatccttg ctgttcccac
     1081 ccatgtcaaa accgaggtgt atgtatgagt gtgggatttg accagtataa gtgcgattgt
     1141 acccggacag gattctatgg agaaaactgc tcaacaccgg aatttttgac aagaataaaa
     1201 ttatttctga aacccactcc aaacacagtg cactacatac ttacccactt caagggattt
     1261 tggaacgttg tgaataacat tcccttcctt cgaaatgcaa ttatgagtta tgtgttgaca
     1321 tccagatcac atttgattga cagtccacca acttacaatg ctgactatgg ctacaaaagc
     1381 tgggaagcct tctctaacct ctcctattat actagagccc ttcctcctgt gcctgatgat
     1441 tgcccgactc ccttgggtgt caaaggtaaa aagcagcttc ctgattcaaa tgagattgtg
     1501 gaaaaattgc ttctaagaag aaagttcatc cctgatcccc agggctcaaa catgatgttt
     1561 gcattctttg cccagcactt cacgcatcag tttttcaaga cagatcataa gcgagggcca
     1621 gctttcacca acgggctggg ccatggggtg gacttaaatc atatttacgg tgaaactctg
     1681 gctagacagc gtaaactgcg ccttttcaag gatggaaaaa tgaaatatca gataattgat
     1741 ggagagatgt atcctcccac agtcaaagat actcaggcag agatgatcta ccctcctcaa
     1801 gtccctgagc atctacggtt tgctgtgggg caggaggtct ttggtctggt gcctggtctg
     1861 atgatgtatg ccacaatctg gctgcgggaa cacaacagag tatgcgatgt gcttaaacag
     1921 gagcatcctg aatggggtga tgagcagttg ttccagacaa gcaggctaat actgatagga
     1981 gagactatta agattgtgat tgaagattat gtgcaacact tgagtggcta tcacttcaaa
     2041 ctgaaatttg acccagaact acttttcaac aaacaattcc agtaccaaaa tcgtattgct
     2101 gctgaattta acaccctcta tcactggcat ccccttctgc ctgacacctt tcaaattcat
     2161 gaccagaaat acaactatca acagtttatc tacaacaact ctatattgct ggaacatgga
     2221 attacccagt ttgttgaatc attcaccagg caaattgctg gcagggttgc tggtggtagg
     2281 aatgttccac ccgcagtaca gaaagtatca caggcttcca ttgaccagag caggcagatg
     2341 aaataccagt cttttaatga gtaccgcaaa cgctttatgc tgaagcccta tgaatcattt
     2401 gaagaactta caggagaaaa ggaaatgtct gcagagttgg aagcactcta tggtgacatc
     2461 gatgctgtgg agctgtatcc tgcccttctg gtagaaaagc ctcggccaga tgccatcttt
     2521 ggtgaaacca tggtagaagt tggagcacca ttctccttga aaggacttat gggtaatgtt
     2581 atatgttctc ctgcctactg gaagccaagc acttttggtg gagaagtggg ttttcaaatc
     2641 atcaacactg cctcaattca gtctctcatc tgcaataacg tgaagggctg tccctttact
     2701 tcattcagtg ttccagatcc agagctcatt aaaacagtca ccatcaatgc aagttcttcc
     2761 cgctccggac tagatgatat caatcccaca gtactactaa aagaacgttc gactgaactg
     2821 taggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa
     2881 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2941 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3001 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3061 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3121 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3181 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3241 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3301 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3361 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3421 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3481 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3541 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3601 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3661 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3721 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3781 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3841 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3901 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3961 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4021 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4081 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4141 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4201 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4261 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4321 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4381 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4441 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4501 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4561 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4621 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4681 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4741 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4801 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4861 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4921 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4981 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5041 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5101 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5161 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5221 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5281 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5341 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5401 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5461 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5521 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5581 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5641 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5701 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5761 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5821 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5881 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5941 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6001 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6061 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6301 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6361 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6421 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6481 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6541 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6601 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6661 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6721 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6781 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6841 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6901 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6961 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7021 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7081 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7141 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7201 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7261 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
