LOCUS       V026558                 8023 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026558
VERSION     V026558
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..3513
                     /label="KEAP1(NM_203500)"
                     /note="KEAP1(NM_203500)"
                     /gene="KEAP1"
     polyA_signal    3590..3814
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3860..4288
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4302..4631
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4698..5492
                     /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    5666..5787
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5836..5852)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5860..5876
                     /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(5884..5914)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5929..5950
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6238..6823)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6994..7854)
                     /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(7855..7959)
                     /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 catgcagcca gatcccaggc ctagcggggc tggggcctgc
     1681 tgccgattcc tgcccctgca gtcacagtgc cctgaggggg caggggacgc ggtgatgtac
     1741 gcctccactg agtgcaaggc ggaggtgacg ccctcccagc atggcaaccg caccttcagc
     1801 tacaccctgg aggatcatac caagcaggcc tttggcatca tgaacgagct gcggctcagc
     1861 cagcagctgt gtgacgtcac actgcaggtc aagtaccagg atgcaccggc cgcccagttc
     1921 atggcccaca aggtggtgct ggcctcatcc agccctgtct tcaaggccat gttcaccaac
     1981 gggctgcggg agcagggcat ggaggtggtg tccattgagg gtatccaccc caaggtcatg
     2041 gagcgcctca ttgaattcgc ctacacggcc tccatctcca tgggcgagaa gtgtgtcctc
     2101 cacgtcatga acggtgctgt catgtaccag atcgacagcg ttgtccgtgc ctgcagtgac
     2161 ttcctggtgc agcagctgga ccccagcaat gccatcggca tcgccaactt cgctgagcag
     2221 attggctgtg tggagttgca ccagcgtgcc cgggagtaca tctacatgca ttttggggag
     2281 gtggccaagc aagaggagtt cttcaacctg tcccactgcc aactggtgac cctcatcagc
     2341 cgggacgacc tgaacgtgcg ctgcgagtcc gaggtcttcc acgcctgcat caactgggtc
     2401 aagtacgact gcgaacagcg acggttctac gtccaggcgc tgctgcgggc cgtgcgctgc
     2461 cactcgttga cgccgaactt cctgcagatg cagctgcaga agtgcgagat cctgcagtcc
     2521 gactcccgct gcaaggacta cctggtcaag atcttcgagg agctcaccct gcacaagccc
     2581 acgcaggtga tgccctgccg ggcgcccaag gtgggccgcc tgatctacac cgcgggcggc
     2641 tacttccgac agtcgctcag ctacctggag gcttacaacc ccagtgacgg cacctggctc
     2701 cggttggcgg acctgcaggt gccgcggagc ggcctggccg gctgcgtggt gggcgggctg
     2761 ttgtacgccg tgggcggcag gaacaactcg cccgacggca acaccgactc cagcgccctg
     2821 gactgttaca accccatgac caatcagtgg tcgccctgcg cccccatgag cgtgccccgt
     2881 aaccgcatcg gggtgggggt catcgatggc cacatctatg ccgtcggcgg ctcccacggc
     2941 tgcatccacc acaacagtgt ggagaggtat gagccagagc gggatgagtg gcacttggtg
     3001 gccccaatgc tgacacgaag gatcggggtg ggcgtggctg tcctcaatcg tctcctttat
     3061 gccgtggggg gctttgacgg gacaaaccgc cttaattcag ctgagtgtta ctacccagag
     3121 aggaacgagt ggcgaatgat cacagcaatg aacaccatcc gaagcggggc aggcgtctgc
     3181 gtcctgcaca actgtatcta tgctgctggg ggctatgatg gtcaggacca gctgaacagc
     3241 gtggagcgct acgatgtgga aacagagacg tggactttcg tagcccccat gaagcaccgg
     3301 cgaagtgccc tggggatcac tgtccaccag gggagaatct acgtccttgg aggctatgat
     3361 ggtcacacgt tcctggacag tgtggagtgt tacgacccag atacagacac ctggagcgag
     3421 gtgacccgaa tgacatcggg ccggagtggg gtgggcgtgg ctgtcaccat ggagccctgc
     3481 cggaagcaga ttgaccagca gaactgtacc tgttgaggat cctgatatcc agcacagtgg
     3541 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3601 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3661 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3721 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3781 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3841 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3901 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3961 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4021 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4081 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4141 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4201 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4261 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4321 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4381 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4441 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4501 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4561 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4621 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4681 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4741 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4801 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4861 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4921 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4981 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5041 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5101 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5161 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5221 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5281 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5341 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5401 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5461 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5521 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5581 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5641 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5701 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5761 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5821 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5881 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5941 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6001 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6061 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6121 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6661 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6901 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6961 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7201 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7261 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7861 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7921 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7981 acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
