LOCUS       V025875                 7921 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025875
VERSION     V025875
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"
     CDS             706..2577
                     /label="KEAP1(NM_012289)"
                     /note="KEAP1(NM_012289)"
                     /gene="KEAP1"
     CDS             2584..2673
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2697..3285
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3351..3399
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3601..4029
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4043..4372
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4439..5233
                     /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    5409..5530
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5579..5595)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5603..5619
                     /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(5627..5657)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5672..5693
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5981..6569)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6740..7600)
                     /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(7601..7705)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgca gccagatccc
      721 aggcctagcg gggctggggc ctgctgccga ttcctgcccc tgcagtcaca gtgccctgag
      781 ggggcagggg acgcggtgat gtacgcctcc actgagtgca aggcggaggt gacgccctcc
      841 cagcatggca accgcacctt cagctacacc ctggaggatc ataccaagca ggcctttggc
      901 atcatgaacg agctgcggct cagccagcag ctgtgtgacg tcacactgca ggtcaagtac
      961 caggatgcac cggccgccca gttcatggcc cacaaggtgg tgctggcctc atccagccct
     1021 gtcttcaagg ccatgttcac caacgggctg cgggagcagg gcatggaggt ggtgtccatt
     1081 gagggtatcc accccaaggt catggagcgc ctcattgaat tcgcctacac ggcctccatc
     1141 tccatgggcg agaagtgtgt cctccacgtc atgaacggtg ctgtcatgta ccagatcgac
     1201 agcgttgtcc gtgcctgcag tgacttcctg gtgcagcagc tggaccccag caatgccatc
     1261 ggcatcgcca acttcgctga gcagattggc tgtgtggagt tgcaccagcg tgcccgggag
     1321 tacatctaca tgcattttgg ggaggtggcc aagcaagagg agttcttcaa cctgtcccac
     1381 tgccaactgg tgaccctcat cagccgggac gacctgaacg tgcgctgcga gtccgaggtc
     1441 ttccacgcct gcatcaactg ggtcaagtac gactgcgaac agcgacggtt ctacgtccag
     1501 gcgctgctgc gggccgtgcg ctgccactcg ttgacgccga acttcctgca gatgcagctg
     1561 cagaagtgcg agatcctgca gtccgactcc cgctgcaagg actacctggt caagatcttc
     1621 gaggagctca ccctgcacaa gcccacgcag gtgatgccct gccgggcgcc caaggtgggc
     1681 cgcctgatct acaccgcggg cggctacttc cgacagtcgc tcagctacct ggaggcttac
     1741 aaccccagtg acggcacctg gctccggttg gcggacctgc aggtgccgcg gagcggcctg
     1801 gccggctgcg tggtgggcgg gctgttgtac gccgtgggcg gcaggaacaa ctcgcccgac
     1861 ggcaacaccg actccagcgc cctggactgt tacaacccca tgaccaatca gtggtcgccc
     1921 tgcgccccca tgagcgtgcc ccgtaaccgc atcggggtgg gggtcatcga tggccacatc
     1981 tatgccgtcg gcggctccca cggctgcatc caccacaaca gtgtggagag gtatgagcca
     2041 gagcgggatg agtggcactt ggtggcccca atgctgacac gaaggatcgg ggtgggcgtg
     2101 gctgtcctca atcgtctcct ttatgccgtg gggggctttg acgggacaaa ccgccttaat
     2161 tcagctgagt gttactaccc agagaggaac gagtggcgaa tgatcacagc aatgaacacc
     2221 atccgaagcg gggcaggcgt ctgcgtcctg cacaactgta tctatgctgc tgggggctat
     2281 gatggtcagg accagctgaa cagcgtggag cgctacgatg tggaaacaga gacgtggact
     2341 ttcgtagccc ccatgaagca ccggcgaagt gccctgggga tcactgtcca ccaggggaga
     2401 atctacgtcc ttggaggcta tgatggtcac acgttcctgg acagtgtgga gtgttacgac
     2461 ccagatacag acacctggag cgaggtgacc cgaatgacat cgggccggag tggggtgggc
     2521 gtggctgtca ccatggagcc ctgccggaag cagattgacc agcagaactg tacctgtctc
     2581 gaggaacaaa aactaatatc agaggaagat ttggaacaga aactaatcag tgaagaagat
     2641 ttagagcaaa agttaatttc cgaggaggac ttatagtgag ggcccgatat ctcgacaatc
     2701 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2761 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2821 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2881 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2941 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3001 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3061 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     3121 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3181 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3241 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag
     3301 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3361 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3421 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3481 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3541 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     3601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3661 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3721 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     3781 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3901 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3961 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4021 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4201 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4321 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4381 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4441 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4501 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4561 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4621 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4681 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4741 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4801 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4861 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4921 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4981 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5041 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5101 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5161 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5221 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     5281 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5341 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5401 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5461 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5521 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5581 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5641 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5701 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5761 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5821 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5881 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5941 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6001 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6061 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6121 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     6181 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6241 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6301 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6361 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6421 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6481 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6541 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6601 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6661 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6721 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6781 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6841 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6901 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6961 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7021 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7081 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7141 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7201 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7261 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7321 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7381 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7441 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7501 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7561 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7621 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7681 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7741 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
     7801 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     7861 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     7921 g
//
