LOCUS       V033539                 7500 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033539
VERSION     V033539
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..2180
                     /label="Pparg(NM_001145366)"
                     /note="Pparg(NM_001145366)"
                     /gene="Pparg"
     CDS             2187..2252
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2276..2864
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2930..2978
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3180..3608
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3622..3951
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4018..4812
                     /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    4988..5109
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5158..5174)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5182..5198
                     /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(5206..5236)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5251..5272
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5560..6148)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6319..7179)
                     /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(7180..7284)
                     /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 ccaccatggt tgacacagag atgccattct
      781 ggcccaccaa cttcggaatc agctctgtgg acctctctgt gatggatgac cactcccatt
      841 cctttgacat caaacccttt accacggttg atttctccag catttctgct ccacactatg
      901 aagacatccc gttcacaaga gctgacccaa tggttgctga ttacaaatat gacctgaagc
      961 tccaagaata ccaaagtgcg atcaaagtag agcctgcgtc cccgccttat tattctgaaa
     1021 aaacccaact ctacaacagg ccacatgaag agccttcaaa ctccctcatg gccatcgagt
     1081 gccgagtctg tggggataaa gcatcaggct tccactatgg agtccatgct tgtgaaggat
     1141 gcaagggttt tttccgaaga accatccgat tgaagcttat ttatgatagg tgtgatctta
     1201 actgtcggat ccacaaaaag agtagaaata aatgtcagta ctgtcggttt cagaagtgcc
     1261 ttgctgtggg gatgtctcac aatgccatca ggtttgggcg aatgccacag gccgagaagg
     1321 agaagctgtt ggcggagatc tccagtgata tcgaccagct gaacccagag tctgctgatc
     1381 tgcgagccct ggcaaagcat ttgtatgact catacataaa gtccttcccg ctgaccaaag
     1441 ccaaggcgag ggcgatcttg acaggaaaga caacagacaa atcaccattt gtcatctacg
     1501 acatgaattc cttaatgatg ggagaagaca aaatcaagtt caaacatatc acccccctgc
     1561 aggagcagag caaagaggtg gccatccgca tttttcaagg gtgccagttt cgatccgtgg
     1621 aagctgtgca agagatcaca gagtatgcca aaaatatccc tggtttcatt aaccttgact
     1681 tgaatgacca agtgactctg ctcaagtatg gtgtccatga gatcatctac accatgctgg
     1741 cctccctgat gaataaagat ggagtcctca tatcagaggg acaaggattc atgaccaggg
     1801 agttcctcaa aagcctgcgg aagccctttg gtgactttat ggagcctaag tttgagtttg
     1861 ctgtgaagtt caatgcactg gaattagatg acagtgactt ggccatattt atagctgtca
     1921 ttattctcag tggagaccgc ccaggcttgc tgaacgtgaa gcccatcgag gacatccaag
     1981 acaacctgct gcaggccctg gaactccagc tgaagctgaa ccacccggag tcctcccagc
     2041 tgttcgccaa ggtgctccag aagatgacag acctcaggca gattgtcaca gagcacgtgc
     2101 agctactgca tgtgatcaag aagacggaga cagatatgag ccttcaccct ctgctccagg
     2161 agatctacaa ggacttgtat ggatccgact acaaagacca tgacggtgat tataaagatc
     2221 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2281 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2341 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2401 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2461 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2521 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2581 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2641 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2701 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2761 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2821 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     2881 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     2941 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3001 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3061 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3121 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3181 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3241 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3301 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3361 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3421 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3481 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3541 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3601 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3661 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3721 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3781 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3841 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3901 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3961 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4021 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4081 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4141 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4201 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4261 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4321 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4381 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4441 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4501 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4561 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4621 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4681 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4741 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4801 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     4861 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4921 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4981 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5041 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5101 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5161 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5221 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5281 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5341 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5401 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5461 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5521 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5581 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5641 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5701 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5761 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5821 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5881 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5941 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6001 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6061 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6121 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6181 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6241 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6301 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6361 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6421 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6481 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6541 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6601 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6661 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6721 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6781 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6841 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6901 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6961 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7021 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7081 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7141 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7201 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7261 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7321 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7381 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7441 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
