LOCUS       V021812                 6878 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021812
VERSION     V021812
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2368
                     /label="Nono(NM_001012356)"
                     /note="Nono(NM_001012356)"
                     /gene="Nono"
     CDS             2375..2440
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2478..2702
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2748..3176
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3190..3519
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3586..4380
                     /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    4554..4675
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4724..4740)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4748..4764
                     /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(4772..4802)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4817..4838
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5126..5711)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5882..6742)
                     /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(6743..6847)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgcagagca ataaaacctt
      961 taacttggag aagcagaatc atactccaag gaagcaccat cagcatcacc accagcagca
     1021 ccatcagcaa cagcagcagc agcagcagca gcagcagcag cagccacccc caccaatacc
     1081 tgcaaatgga cagcaggcca gcagccagaa tgaaggcttg actattgacc tgaagaattt
     1141 taggaaacca ggagagaaga cctttaccca acgtagccgt ctctttgtgg gcaatcttcc
     1201 ccctgacatc actgaggagg aaatgaggaa actttttgag aaatatggga aagcaggcga
     1261 agttttcatt cataaggata aaggctttgg ctttattcgc ttggaaacac gaaccctagc
     1321 ggaaattgcc aaagtggagc tggacaacat gccactccgt ggaaagcagc tgcgagtgcg
     1381 ctttgcttgt catagtgcat cccttacggt tcgcaacctt cctcagtacg tgtccaacga
     1441 actgctggaa gaagcctttt ctgtgtttgg ccaggtggag agggctgtag tcattgtgga
     1501 tgaccgagga aggccctcgg ggaaaggcat tgttgagttc tcagggaagc cagctgctcg
     1561 gaaagctctg gacagatgca gtgaaggctc cttcttgcta accacatttc ctcggcctgt
     1621 gactgtggag cctatggacc agttagatga tgaagaggga cttccggaga agttggttat
     1681 aaaaaaccag caatttcaca aggagagaga acagccaccc agatttgcac agcctgggtc
     1741 ctttgagtat gagtatgcca tgcgctggaa ggctctcatt gagatggaga agcaacagca
     1801 ggatcaagtg gaccgaaaca tcaaggaggc tcgtgagaag ctggagatgg agatggaggc
     1861 agcacgccat gagcaccagg ttatgctaat gaggcaggat ttgatgagac gtcaagaaga
     1921 gcttcgaaga atggaggagc tgcataacca agaggttcag aaacgaaagc agttagaact
     1981 caggcaggaa gaggaacgca ggcgccgtga ggaagagatg cggcgacagc aagaagaaat
     2041 gatgcgacgg cagcaggaag gattcaaggg aaccttccct gatgcgagag aacaagagat
     2101 acggatgggc caaatggcta tgggaggtgc catgggcata aacaatagag gcgccatgcc
     2161 ccctgctcct gtgccacctg gtactccagc tcctccagga cctgctgcta tgatgccaga
     2221 tggaaccctt ggattgactc caccaacaac tgaacgtttt ggccaagctg ctacaatgga
     2281 aggaattgga gcaattggtg gaactcctcc tgcattcaac cgtccagctc ctggagctga
     2341 atttgctcca aataaacgcc gccgatatct cgaggactac aaagaccatg acggtgatta
     2401 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac
     2461 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2521 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2581 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2641 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2701 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2761 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2821 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2881 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2941 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3001 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3061 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3121 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3181 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3241 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3301 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3361 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3421 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3481 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3541 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3601 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3661 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3721 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3781 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3841 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3901 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3961 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4021 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4081 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4141 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4201 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4261 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4321 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4381 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4441 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4501 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4561 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4621 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4681 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4741 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4801 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4861 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4921 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4981 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5041 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5101 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5161 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5221 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5281 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5341 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5401 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5461 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5521 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5581 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5641 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5701 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5761 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5821 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5881 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5941 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6001 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6061 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6121 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6181 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6241 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6301 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6361 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6421 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6481 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6541 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6601 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6661 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6721 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6781 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6841 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
