LOCUS       V039704                 8085 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039704
VERSION     V039704
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             930..956
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             960..986
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             990..1016
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1017..2888
                     /label="KEAP1(NM_203500)"
                     /note="KEAP1(NM_203500)"
                     /gene="KEAP1"
     CDS             2928..3647
                     /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    3685..3909
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3955..4383
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4397..4726
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4793..5587
                     /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    5761..5882
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5931..5947)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5955..5971
                     /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(5979..6009)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6024..6045
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6333..6918)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7089..7949)
                     /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(7950..8054)
                     /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 gtttaaagcc actgggatcc gccaccatgt acccatacga cgtacctgat tatgcaactt
      961 atccttatga tgtacctgat tatgctagct atccttatga tgtcccagac tacgcaatgc
     1021 agccagatcc caggcctagc ggggctgggg cctgctgccg attcctgccc ctgcagtcac
     1081 agtgccctga gggggcaggg gacgcggtga tgtacgcctc cactgagtgc aaggcggagg
     1141 tgacgccctc ccagcatggc aaccgcacct tcagctacac cctggaggat cataccaagc
     1201 aggcctttgg catcatgaac gagctgcggc tcagccagca gctgtgtgac gtcacactgc
     1261 aggtcaagta ccaggatgca ccggccgccc agttcatggc ccacaaggtg gtgctggcct
     1321 catccagccc tgtcttcaag gccatgttca ccaacgggct gcgggagcag ggcatggagg
     1381 tggtgtccat tgagggtatc caccccaagg tcatggagcg cctcattgaa ttcgcctaca
     1441 cggcctccat ctccatgggc gagaagtgtg tcctccacgt catgaacggt gctgtcatgt
     1501 accagatcga cagcgttgtc cgtgcctgca gtgacttcct ggtgcagcag ctggacccca
     1561 gcaatgccat cggcatcgcc aacttcgctg agcagattgg ctgtgtggag ttgcaccagc
     1621 gtgcccggga gtacatctac atgcattttg gggaggtggc caagcaagag gagttcttca
     1681 acctgtccca ctgccaactg gtgaccctca tcagccggga cgacctgaac gtgcgctgcg
     1741 agtccgaggt cttccacgcc tgcatcaact gggtcaagta cgactgcgaa cagcgacggt
     1801 tctacgtcca ggcgctgctg cgggccgtgc gctgccactc gttgacgccg aacttcctgc
     1861 agatgcagct gcagaagtgc gagatcctgc agtccgactc ccgctgcaag gactacctgg
     1921 tcaagatctt cgaggagctc accctgcaca agcccacgca ggtgatgccc tgccgggcgc
     1981 ccaaggtggg ccgcctgatc tacaccgcgg gcggctactt ccgacagtcg ctcagctacc
     2041 tggaggctta caaccccagt gacggcacct ggctccggtt ggcggacctg caggtgccgc
     2101 ggagcggcct ggccggctgc gtggtgggcg ggctgttgta cgccgtgggc ggcaggaaca
     2161 actcgcccga cggcaacacc gactccagcg ccctggactg ttacaacccc atgaccaatc
     2221 agtggtcgcc ctgcgccccc atgagcgtgc cccgtaaccg catcggggtg ggggtcatcg
     2281 atggccacat ctatgccgtc ggcggctccc acggctgcat ccaccacaac agtgtggaga
     2341 ggtatgagcc agagcgggat gagtggcact tggtggcccc aatgctgaca cgaaggatcg
     2401 gggtgggcgt ggctgtcctc aatcgtctcc tttatgccgt ggggggcttt gacgggacaa
     2461 accgccttaa ttcagctgag tgttactacc cagagaggaa cgagtggcga atgatcacag
     2521 caatgaacac catccgaagc ggggcaggcg tctgcgtcct gcacaactgt atctatgctg
     2581 ctgggggcta tgatggtcag gaccagctga acagcgtgga gcgctacgat gtggaaacag
     2641 agacgtggac tttcgtagcc cccatgaagc accggcgaag tgccctgggg atcactgtcc
     2701 accaggggag aatctacgtc cttggaggct atgatggtca cacgttcctg gacagtgtgg
     2761 agtgttacga cccagataca gacacctgga gcgaggtgac ccgaatgaca tcgggccgga
     2821 gtggggtggg cgtggctgtc accatggagc cctgccggaa gcagattgac cagcagaact
     2881 gtacctgtgc ctcgagtggc ggtggaggaa gcggcggagg tggatctatg gtgagcaagg
     2941 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     3001 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     3061 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3121 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     3181 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     3241 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     3301 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     3361 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     3421 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     3481 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     3541 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3601 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtagtct agagggcccg
     3661 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3721 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3781 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3841 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3901 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3961 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4021 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4081 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4141 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4201 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4261 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4321 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4381 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4441 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4501 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4561 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4621 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4681 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4741 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4801 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4861 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4921 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4981 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5041 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5101 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5161 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5221 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5281 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     5341 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5401 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5461 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5521 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5581 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5641 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5701 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5761 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5821 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5881 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5941 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6001 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6121 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6841 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     6901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6961 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7021 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7081 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7201 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7381 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7441 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7501 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7561 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7621 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7681 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7741 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7801 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7861 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7921 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8041 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
