LOCUS       V031587                 8560 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031587
VERSION     V031587
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1642..4050
                     /label="PPARGC1A(NM_001330751)"
                     /note="PPARGC1A(NM_001330751)"
                     /gene="PPARGC1A"
     polyA_signal    4127..4351
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4397..4825
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4839..5168
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5235..6029
                     /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    6203..6324
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6373..6389)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6397..6413
                     /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(6421..6451)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6466..6487
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6775..7360)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7531..8391)
                     /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(8392..8496)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggatgag acctccccaa ggctggaaga agactggaaa
     1681 aaagtacttc agcgagaagc aggctggcag tgtgctgctc tggttggtga agaccagcct
     1741 ctttgcccag atcttcctga acttgatctt tctgaactag atgtgaacga cttggataca
     1801 gacagctttc tgggtggact caagtggtgc agtgaccaat cagaaataat atccaatcag
     1861 tacaacaatg agccttcaaa catatttgag aagatagatg aagagaatga ggcaaacttg
     1921 ctagcagtcc tcacagagac actagacagt ctccctgtgg atgaagacgg attgccctca
     1981 tttgatgcgc tgacagatgg agacgtgacc actgacaatg aggctagtcc ttcctccatg
     2041 cctgacggca cccctccacc ccaggaggca gaagagccgt ctctacttaa gaagctctta
     2101 ctggcaccag ccaacactca gctaagttat aatgaatgca gtggtctcag tacccagaac
     2161 catgcaaatc acaatcacag gatcagaaca aaccctgcaa ttgttaagac tgagaattca
     2221 tggagcaata aagcgaagag tatttgtcaa cagcaaaagc cacaaagacg tccctgctcg
     2281 gagcttctca aatatctgac cacaaacgat gaccctcctc acaccaaacc cacagagaac
     2341 agaaacagca gcagagacaa atgcacctcc aaaaagaagt cccacacaca gtcgcagtca
     2401 caacacttac aagccaaacc aacaacttta tctcttcctc tgaccccaga gtcaccaaat
     2461 gaccccaagg gttccccatt tgagaacaag actattgaac gcaccttaag tgtggaactc
     2521 tctggaactg caggcctaac tccacccacc actcctcctc ataaagccaa ccaagataac
     2581 ccttttaggg cttctccaaa gctgaagtcc tcttgcaaga ctgtggtgcc accaccatca
     2641 aagaagccca ggtacagtga gtcttctggt acacaaggca ataactccac caagaaaggg
     2701 ccggagcaat ccgagttgta tgcacaactc agcaagtcct cagtcctcac tggtggacac
     2761 gaggaaagga agaccaagcg gcccagtctg cggctgtttg gtgaccatga ctattgccag
     2821 tcaattaatt ccaaaacaga aatactcatt aatatatcac aggagctcca agactctaga
     2881 caactagaaa ataaagatgt ctcctctgat tggcaggggc agatttgttc ttccacagat
     2941 tcagaccagt gctacctgag agagactttg gaggcaagca agcaggtctc tccttgcagc
     3001 acaagaaaac agctccaaga ccaggaaatc cgagccgagc tgaacaagca cttcggtcat
     3061 cccagtcaag ctgtttttga cgacgaagca gacaagaccg gtgaactgag ggacagtgat
     3121 ttcagtaatg aacaattctc caaactacct atgtttataa attcaggact agccatggat
     3181 ggcctgtttg atgacagcga agatgaaagt gataaactga gctacccttg ggatggcacg
     3241 caatcctatt cattgttcaa tgtgtctcct tcttgttctt cttttaactc tccatgtaga
     3301 gattctgtgt caccacccaa atccttattt tctcaaagac cccaaaggat gcgctctcgt
     3361 tcaaggtcct tttctcgaca caggtcgtgt tcccgatcac catattccag gtcaagatca
     3421 aggtctccag gcagtagatc ctcttcaaga tcctgctatt actatgagtc aagccactac
     3481 agacaccgca cgcaccgaaa ttctcccttg tatgtgagat cacgttcaag atcgccctac
     3541 agccgtcggc ccaggtatga cagctacgag gaatatcagc acgagaggct gaagagggaa
     3601 gaatatcgca gagagtatga gaagcgagag tctgagaggg ccaagcaaag ggagaggcag
     3661 aggcagaagg caattgaaga gcgccgtgtg atttatgtcg gtaaaatcag acctgacaca
     3721 acacggacag aactgaggga ccgttttgaa gtttttggtg aaattgagga gtgcacagta
     3781 aatctgcggg atgatggaga cagctatggt ttcattacct accgttatac ctgtgatgct
     3841 tttgctgctc ttgaaaatgg atacactttg cgcaggtcaa acgaaactga ctttgagctg
     3901 tacttttgtg gacgcaagca atttttcaag tctaactatg cagacctaga ttcaaactca
     3961 gatgactttg accctgcttc caccaagagc aagtatgact ctctggattt tgatagttta
     4021 ctgaaagaag ctcagagaag cttgcgcagg taaggatcct gatatccagc acagtggcgg
     4081 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     4141 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4201 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4261 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4321 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4381 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4441 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4501 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4561 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4621 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4681 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4741 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4801 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4921 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4981 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5041 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5101 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5161 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5221 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5281 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5341 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5401 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5461 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5521 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5581 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5641 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5701 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5761 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5821 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5881 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5941 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6001 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     6061 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     6121 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6181 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6241 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6301 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6361 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6421 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6481 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6541 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6601 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6661 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6721 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6781 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6841 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6901 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6961 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7021 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7081 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7141 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7201 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7261 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     7321 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7381 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7441 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7501 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7561 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7621 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7681 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7741 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7801 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7861 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7921 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7981 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8041 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8101 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8161 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8221 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8281 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8341 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8401 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8461 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8521 tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
