LOCUS       V028047                 7277 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028047
VERSION     V028047
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"
     CDS             905..1969
                     /label="PON1(NM_000446)"
                     /note="PON1(NM_000446)"
                     /gene="PON1"
     CDS             1982..2047
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2057..2113
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             2126..2845
                     /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"
     polyA_signal    2877..3101
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3147..3575
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3589..3918
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3985..4779
                     /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    4953..5074
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5123..5139)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5147..5163
                     /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(5171..5201)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5216..5237
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5525..6110)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6281..7141)
                     /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(7142..7246)
                     /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 caccatggcg aagctgattg cgctcaccct cttggggatg ggactggcac tcttcaggaa
      961 ccaccagtct tcttaccaaa cacgacttaa tgctctccga gaggtacaac ccgtagaact
     1021 tcctaactgt aatttagtta aaggaatcga aactggctct gaagacttgg agatactgcc
     1081 taatggactg gctttcatta gctctggatt aaagtatcct ggaataaaga gcttcaaccc
     1141 caacagtcct ggaaaaatac ttctgatgga cctgaatgaa gaagatccaa cagtgttgga
     1201 attggggatc actggaagta aatttgatgt atcttcattt aaccctcatg ggattagcac
     1261 attcacagat gaagataatg ccatgtacct cctggtggtg aaccatccag atgccaagtc
     1321 cacagtggag ttgtttaaat ttcaagaaga agaaaaatcg cttttgcatc taaaaaccat
     1381 cagacataaa cttctgccta atttgaatga tattgttgct gtgggacctg agcactttta
     1441 tggcacaaat gatcactatt ttcttgaccc ctacttacaa tcctgggaga tgtatttggg
     1501 tttagcgtgg tcgtatgttg tctactatag tccaagtgaa gttcgagtgg tggcagaagg
     1561 atttgatttt gctaatggaa tcaacatttc acccgatggc aagtatgtct atatagctga
     1621 gttgctggct cataagattc atgtgtatga aaagcatgct aattggactt taactccatt
     1681 gaagtccctt gactttaata ccctcgtgga taacatatct gtggatcctg agacaggaga
     1741 cctttgggtt ggatgccatc ccaatggcat gaaaatcttc ttctatgact cagagaatcc
     1801 tcctgcatca gaggtgcttc gaatccagaa cattctaaca gaagaaccta aagtgacaca
     1861 ggtttatgca gaaaatggca cagtgttgca aggcagtaca gttgcctctg tgtacaaagg
     1921 gaaactgctg attggcacag tgtttcacaa agctctttac tgtgagctcc tcgagtctag
     1981 agactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2041 tgacaaggga agcggagcca ctaacttctc cctgttgaaa caagcagggg atgtcgaaga
     2101 gaatcccggg ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     2161 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2221 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2281 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     2341 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2401 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2461 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2521 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     2581 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     2641 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2701 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     2761 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2821 cggcatggac gagctgtaca agtaagggcc cgtttaaacc cgctgatcag cctcgactgt
     2881 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2941 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3001 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3061 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3121 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3301 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3361 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3481 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3541 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3601 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3721 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3781 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3841 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3901 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3961 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4021 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4081 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4141 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4201 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4261 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4321 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4381 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4441 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4501 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4561 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4621 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4681 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4741 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4801 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4861 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4921 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4981 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5041 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5101 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5161 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5221 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5281 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5341 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5521 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5701 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5941 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6061 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6121 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6181 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6241 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6301 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6361 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6421 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6481 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6541 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6601 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6661 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6721 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6781 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6841 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6901 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6961 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7021 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7081 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7141 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7201 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7261 aagtgccacc tgacgtc
//
