LOCUS       V020009                 7736 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020009
VERSION     V020009
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..2416
                     /label="Nox4(NM_053524)"
                     /note="Nox4(NM_053524)"
                     /gene="Nox4"
     CDS             2423..2488
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2512..3100
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3166..3214
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3416..3844
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3858..4187
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4254..5048
                     /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    5224..5345
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5394..5410)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5418..5434
                     /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(5442..5472)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5487..5508
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5796..6384)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6555..7415)
                     /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(7416..7520)
                     /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 ccatggcgct gtcctggagg agctggctgg ccaacgaagg
      721 ggttaaacac ctctgtctgc ttgtttggct gtccctaaat gtcctgcttt tctggaaaac
      781 cttcctgctg tacaaccaag ggccagaata ctactacatc caccagatgt tgggcctagg
      841 attgtgtttg agcagagctt ctgcatctgt cctgaacctc aactgcagcc tgatcctttt
      901 acccatgtgc cgcacagtcc tggcttacct tcgcggatca cagaaggtcc ctagcaggag
      961 aacaagaaga ttgttggaca aaagcaagac tctacatatc acctgtggca taactatttg
     1021 tattttctca ggtgtgcatg tagctgccca cttggtgaac gccctgaact tctcagtgaa
     1081 ctatagtgaa catttccttg cactgaatgc agcaagatac cagaatgagg atcccagaaa
     1141 gcttctcttc acaactgttc cgggcctgac aggtgtctgc atggtggtgg tattgttcct
     1201 catggttaca gcttctacct atgcaataag agtttctaat tatgatatct tctggtatac
     1261 tcacaacctc ttctttgtct tctacatgct gctgctgctg catgtttcgg gtggcttgtt
     1321 gaagtatcaa accaatttag acactcaccc tcctggctgt atcagtctta accggacccc
     1381 atctcagaat atgtccatag cagactacgt ctcagaacat tttcatggat ctttgcctgg
     1441 agggttttca aaattagaag atcattacca gaaaacactg gtgaagattt gcctggaaga
     1501 acccaagttc caagctcatt tcccacagac ctggatttgg atttctggac ctttgtgcct
     1561 atactgtgct gagagacttt accgatgcat ccggagcaac aaacctgtca ccattatctc
     1621 agtaatcaat catccctcag atgtcatgga actccgtatg atcaaagaaa actttaaagc
     1681 aagacctggc cagtatatta ttctacattg tcccagtgta tcagcattag aaaaccaccc
     1741 atttactctc acaatgtgtc ctactgaaac caaagcaaca tttggtgtcc actttaaagt
     1801 agtaggagac tggacagaaa gattccgaga tttactactg cctccatcaa gccaagattc
     1861 tgagattctg cccttcattc aatctagaaa ctaccccaag ttatacattg atggcccatt
     1921 tggaagtcca tttgaggagt cactgaacta tgaagttagt ctgtgtgtgg ctggaggcat
     1981 tggggtcact ccgtttgcat cgatactaaa cactctactg gatgactgga aaccatacaa
     2041 gctaagaaga ctgtatttta tctgggtctg cagagacatc caatcattcc agtggtttgc
     2101 agacttgctc tatgtgctgc ataacaagtt ttggcaagaa aacagacctg actttgtgaa
     2161 catccagctg tacctcagtc aaacagatgg gatacagaag ataattggag aaaaatacca
     2221 cacattgaat tctagacttt ttattgggcg tcctcggtgg aagcttttat ttgatgaaat
     2281 agcaaaatgt aacagaggga aaacagttgg agttttctgc tgtggaccca gttctatttc
     2341 caagactctt cataatttga gtaaccggaa caactcatat gggacaaaat ttgaatacaa
     2401 taaagaatct ttcagcctcg aggactacaa agaccatgac ggtgattata aagatcatga
     2461 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct
     2521 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2581 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2641 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2701 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2761 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2821 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2881 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2941 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3001 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3061 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     3121 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     3181 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     3241 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     3301 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     3361 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     3421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3481 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3541 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     3601 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3721 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3781 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3841 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3961 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4021 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4081 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4141 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4201 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4261 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4321 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4381 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4441 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4501 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4561 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4621 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4681 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4741 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4801 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4861 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4921 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4981 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5041 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     5101 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5161 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5221 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5281 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5341 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5401 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5461 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5521 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5581 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5641 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5701 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5761 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5821 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5881 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5941 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     6001 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6061 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6121 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6181 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6241 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6301 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6361 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6421 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6481 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6541 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6601 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6661 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6721 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6781 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6841 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6901 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6961 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7021 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7081 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7141 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7201 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7261 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7321 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7381 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7441 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7501 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7561 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     7621 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     7681 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
