LOCUS       V020096                 8084 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020096
VERSION     V020096
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             683..2764
                     /label="NFE2L3(NM_004289)"
                     /note="NFE2L3(NM_004289)"
                     /gene="NFE2L3"
     CDS             2771..2836
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2860..3448
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3514..3562
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3764..4192
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4206..4535
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4602..5396
                     /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    5572..5693
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5742..5758)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5766..5782
                     /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(5790..5820)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5835..5856
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6144..6732)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6903..7763)
                     /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(7764..7868)
                     /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 ggatccgcca ccatgaagca cctgaagcgg tggtggtcgg ccggcggcgg
      721 cctcctgcac ctcaccctcc tgctgagctt ggcggggctc cgcgtagacc tagatcttta
      781 cctgctgctg ccgccgccca ccctgctgca ggacgagctg ctgttcctgg gcggcccggc
      841 cagctccgcc tacgcgctca gccccttctc ggcctcggga gggtgggggc gcgcgggcca
      901 cttgcacccc aagggccggg agctggaccc tgccgcgccg cccgagggcc agctgctccg
      961 ggaggtgcgc gcgctcgggg tccccttcgt ccctcgcacc agcgtggatg catggctggt
     1021 gcacagcgtg gctgccggga gcgcggacga ggcccacggg ctgctcggcg ccgccgccgc
     1081 ctcgtccacc ggaggagccg gcgccagcgt ggacggcggc agccaggctg tgcagggggg
     1141 cggcggggac ccccgagcgg ctcggagtgg ccccttggac gccggggaag aggagaaggc
     1201 acccgcggaa ccgacggctc aggtgccgga cgctggcgga tgtgcgagcg aggagaatgg
     1261 ggtactaaga gaaaagcacg aagctgtgga tcatagttcc cagcatgagg aaaatgaaga
     1321 aagggtgtca gcccagaagg agaactcact tcagcagaat gatgatgatg aaaacaaaat
     1381 agcagagaaa cctgactggg aggcagaaaa gaccactgaa tctagaaatg agagacatct
     1441 gaatgggaca gatacttctt tctctctgga agacttattc cagttgcttt catcacagcc
     1501 tgaaaattca ctggagggca tctcattggg agatattcct cttccaggca gtatcagtga
     1561 tggcatgaat tcttcagcac attatcatgt aaacttcagc caggctataa gtcaggatgt
     1621 gaatcttcat gaggccatct tgctttgtcc caacaataca tttagaagag atccaacagc
     1681 aaggacttca cagtcacaag aaccatttct gcagttaaat tctcatacca ccaatcctga
     1741 gcaaaccctt cctggaacta atttgacagg atttctttca ccggttgaca atcatatgag
     1801 gaatctaaca agccaagacc tactgtatga ccttgacata aatatatttg atgagataaa
     1861 cttaatgtca ttggccacag aagacaactt tgatccaatc gatgtttctc agctttttga
     1921 tgaaccagat tctgattctg gcctttcttt agattcaagt cacaataata cctctgtcat
     1981 caagtctaat tcctctcact ctgtgtgtga tgaaggtgct ataggttatt gcactgacca
     2041 tgaatctagt tcccatcatg acttagaagg tgctgtaggt ggctactacc cagaacccag
     2101 taagctttgt cacttggatc aaagtgattc tgatttccat ggagatctta catttcaaca
     2161 cgtatttcat aaccacactt accacttaca gccaactgca ccagaatcta cttctgaacc
     2221 ttttccgtgg cctgggaagt cacagaagat aaggagtaga taccttgaag acacagatag
     2281 aaacttgagc cgtgatgaac agcgtgctaa agctttgcat atcccttttt ctgtagatga
     2341 aattgtcggc atgcctgttg attctttcaa tagcatgtta agtagatatt atctgacaga
     2401 cctacaagtc tcacttatcc gtgacatcag acgaagaggg aaaaataaag ttgctgcgca
     2461 gaactgtcgt aaacgcaaat tggacataat tttgaattta gaagatgatg tatgtaactt
     2521 gcaagcaaag aaggaaactc ttaagagaga gcaagcacaa tgtaacaaag ctattaacat
     2581 aatgaaacag aaactgcatg acctttatca tgatattttt agtagattaa gagatgacca
     2641 aggtaggcca gtcaatccca accactatgc tctccagtgt acccatgatg gaagtatctt
     2701 gatagtaccc aaagaactgg tggcctcagg ccacaaaaag gaaacccaaa agggaaagag
     2761 aaagctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     2821 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     2881 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2941 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3001 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3061 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3121 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3181 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3241 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3301 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3361 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3421 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     3481 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     3541 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     3601 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3661 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     3721 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3781 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3841 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3901 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     3961 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4021 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4081 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     4141 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4201 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4261 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4321 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4381 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4441 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4501 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4561 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4621 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4681 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4741 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4801 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4861 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4921 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4981 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5041 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5101 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5161 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5221 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5281 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5341 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5401 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5461 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5521 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5581 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5641 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5701 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5761 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5821 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5881 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5941 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6001 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6061 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6121 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6181 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6241 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6301 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     6361 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6421 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6481 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6541 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6601 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6661 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6721 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6781 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6841 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6901 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6961 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7021 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7081 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7141 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7201 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7261 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7321 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7381 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7441 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7501 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7561 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7621 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7681 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7741 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7801 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7861 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     7921 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     7981 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     8041 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
