LOCUS       V031950                 8634 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031950
VERSION     V031950
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             744..1451
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1500..3314
                     /label="NFE2L2(NM_006164)"
                     /note="NFE2L2(NM_006164)"
                     /gene="NFE2L2"
     CDS             3321..3386
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3410..3998
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4064..4112
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4314..4742
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4756..5085
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5152..5946
                     /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    6122..6243
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6292..6308)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6316..6332
                     /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(6340..6370)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6385..6406
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6694..7282)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7453..8313)
                     /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(8314..8418)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca
      781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca
     1501 tgatggactt ggagctgccg ccgccgggac tcccgtccca gcaggacatg gatttgattg
     1561 acatactttg gaggcaagat atagatcttg gagtaagtcg agaagtattt gacttcagtc
     1621 agcgacggaa agagtatgag ctggaaaaac agaaaaaact tgaaaaggaa agacaagaac
     1681 aactccaaaa ggagcaagag aaagcctttt tcgctcagtt acaactagat gaagagacag
     1741 gtgaatttct cccaattcag ccagcccagc acatccagtc agaaaccagt ggatctgcca
     1801 actactccca ggttgcccac attcccaaat cagatgcttt gtactttgat gactgcatgc
     1861 agcttttggc gcagacattc ccgtttgtag atgacaatga ggtttcttcg gctacgtttc
     1921 agtcacttgt tcctgatatt cccggtcaca tcgagagccc agtcttcatt gctactaatc
     1981 aggctcagtc acctgaaact tctgttgctc aggtagcccc tgttgattta gacggtatgc
     2041 aacaggacat tgagcaagtt tgggaggagc tattatccat tcctgagtta cagtgtctta
     2101 atattgaaaa tgacaagctg gttgagacta ccatggttcc aagtccagaa gccaaactga
     2161 cagaagttga caattatcat ttttactcat ctataccctc aatggaaaaa gaagtaggta
     2221 actgtagtcc acattttctt aatgcttttg aggattcctt cagcagcatc ctctccacag
     2281 aagaccccaa ccagttgaca gtgaactcat taaattcaga tgccacagtc aacacagatt
     2341 ttggtgatga attttattct gctttcatag ctgagcccag tatcagcaac agcatgccct
     2401 cacctgctac tttaagccat tcactctctg aacttctaaa tgggcccatt gatgtttctg
     2461 atctatcact ttgcaaagct ttcaaccaaa accaccctga aagcacagca gaattcaatg
     2521 attctgactc cggcatttca ctaaacacaa gtcccagtgt ggcatcacca gaacactcag
     2581 tggaatcttc cagctatgga gacacactac ttggcctcag tgattctgaa gtggaagagc
     2641 tagatagtgc ccctggaagt gtcaaacaga atggtcctaa aacaccagta cattcttctg
     2701 gggatatggt acaacccttg tcaccatctc aggggcagag cactcacgtg catgatgccc
     2761 aatgtgagaa cacaccagag aaagaattgc ctgtaagtcc tggtcatcgg aaaaccccat
     2821 tcacaaaaga caaacattca agccgcttgg aggctcatct cacaagagat gaacttaggg
     2881 caaaagctct ccatatccca ttccctgtag aaaaaatcat taacctccct gttgttgact
     2941 tcaacgaaat gatgtccaaa gagcagttca atgaagctca acttgcatta attcgggata
     3001 tacgtaggag gggtaagaat aaagtggctg ctcagaattg cagaaaaaga aaactggaaa
     3061 atatagtaga actagagcaa gatttagatc atttgaaaga tgaaaaagaa aaattgctca
     3121 aagaaaaagg agaaaatgac aaaagccttc acctactgaa aaaacaactc agcaccttat
     3181 atctcgaagt tttcagcatg ctacgtgatg aagatggaaa accttattct cctagtgaat
     3241 actccctgca gcaaacaaga gatggcaatg ttttccttgt tcccaaaagt aagaagccag
     3301 atgttaagaa aaacctcgag gactacaaag accatgacgg tgattataaa gatcatgaca
     3361 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     3421 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3481 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3541 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3601 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3661 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3721 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3781 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3841 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3901 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3961 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     4021 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4081 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4141 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4201 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4261 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     4321 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4381 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4441 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4501 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4561 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4621 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4681 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4741 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4801 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4861 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4921 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4981 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     5041 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5101 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     5161 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5221 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5281 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5341 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5401 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5461 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5521 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5581 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5641 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5701 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5761 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5821 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5881 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5941 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     6001 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6061 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6121 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6181 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6241 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6301 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6361 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6421 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6481 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6541 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6601 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6661 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6721 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6781 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6841 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6901 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6961 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7021 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7081 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7141 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7201 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7321 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8281 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8401 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8461 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8521 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8581 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
