LOCUS       V022442                 7829 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022442
VERSION     V022442
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..3319
                     /label="UHRF1(NM_001290051)"
                     /note="UHRF1(NM_001290051)"
                     /gene="UHRF1"
     CDS             3326..3391
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3429..3653
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3699..4127
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4141..4470
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4537..5331
                     /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    5505..5626
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5675..5691)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5699..5715
                     /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(5723..5753)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5768..5789
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6077..6662)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6833..7693)
                     /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(7694..7798)
                     /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 atgtggatcc aggttcggac
      961 catggacggg aggcagaccc acacggtgga ctcgctgtcc aggctgacca aggtggagga
     1021 gctgaggcgg aagatccagg agctgttcca cgtggagcca ggcctgcaga ggctgttcta
     1081 caggggcaaa cagatggagg acggccatac cctcttcgac tacgaggtcc gcctgaatga
     1141 caccatccag ctcctggtcc gccagagcct cgtgctcccc cacagcacca aggagcggga
     1201 ctccgagctc tccgacaccg actccggctg ctgcctgggc cagagtgagt cagacaagtc
     1261 ctccacccac ggtgaggcgg ccgccgagac tgacagcagg ccagccgatg aggacatgtg
     1321 ggatgagacg gaattggggc tgtacaaggt caatgagtac gtcgatgctc gggacacgaa
     1381 catgggggcg tggtttgagg cgcaggtggt cagggtgacg cggaaggccc cctcccggga
     1441 cgagccctgc agctccacgt ccaggccggc gctggaggag gacgtcattt accacgtgaa
     1501 atacgacgac tacccggaga acggcgtggt ccagatgaac tccagggacg tccgagcgcg
     1561 cgcccgcacc atcatcaagt ggcaggacct ggaggtgggc caggtggtca tgctcaacta
     1621 caaccccgac aaccccaagg agcggggctt ctggtacgac gcggagatct ccaggaagcg
     1681 cgagaccagg acggcgcggg aactctacgc caacgtggtg ctgggggatg attctctgaa
     1741 cgactgtcgg atcatcttcg tggacgaagt cttcaagatt gagcggccgg gtgaagggag
     1801 ccccatggtt gacaacccca tgagacggaa gagcgggccg tcctgcaagc actgcaagga
     1861 cgacgtgaac agactctgcc gggtctgcgc ctgccacctg tgcgggggcc ggcaggaccc
     1921 cgacaagcag ctcatgtgcg atgagtgcga catggccttc cacatctact gcctggaccc
     1981 gcccctcagc agtgttccca gcgaggacga gtggtactgc cctgagtgcc ggaatgatgc
     2041 cagcgaggtg gtactggcgg gagagcggct gagagagagc aagaagaagg cgaagatggc
     2101 ctcggccaca tcgtcctcac agcgggactg gggcaagggc atggcctgtg tgggccgcac
     2161 caaggaatgt accatcgtcc cgtccaacca ctacggaccc atcccgggga tccccgtggg
     2221 caccatgtgg cggttccgag tccaggtcag cgagtcgggt gtccatcggc cccacgtggc
     2281 tggcatacac ggccggagca acgacggagc gtactcccta gtcctggcgg ggggctatga
     2341 ggatgatgtg gaccatggga attttttcac atacacgggt agtggtggtc gagatctttc
     2401 cggcaacaag aggaccgcgg aacagtcttg tgatcagaaa ctcaccaaca ccaacagggc
     2461 gctggctctc aactgctttg ctcccatcaa tgaccaagaa ggggccgagg ccaaggactg
     2521 gcggtcgggg aagccggtca gggtggtgcg caatgtcaag ggtggcaaga atagcaagta
     2581 cgcccccgct gagggcaacc gctacgatgg catctacaag gttgtgaaat actggcccga
     2641 gaaggggaag tccgggtttc tcgtgtggcg ctaccttctg cggagggacg atgatgagcc
     2701 tggcccttgg acgaaggagg ggaaggaccg gatcaagaag ctggggctga ccatgcagta
     2761 tccagaaggc tacctggaag ccctggccaa ccgagagcga gagaaggaga acagcaagag
     2821 ggaggaggag gagcagcagg aggggggctt cgcgtccccc aggacgggca agggcaagtg
     2881 gaagcggaag tcggcaggag gtggcccgag cagggccggg tccccgcgcc ggacatccaa
     2941 gaaaaccaag gtggagccct acagtctcac ggcccagcag agcagcctca tcagagagga
     3001 caagagcaac gccaagctgt ggaatgaggt cctggcgtca ctcaaggacc ggccggcgag
     3061 cggcagcccg ttccagttgt tcctgagtaa agtggaggag acgttccagt gtatctgctg
     3121 tcaggagctg gtgttccggc ccatcacgac cgtgtgccag cacaacgtgt gcaaggactg
     3181 cctggacaga tcctttcggg cacaggtgtt cagctgccct gcctgccgct acgacctggg
     3241 ccgcagctat gccatgcagg tgaaccagcc tctgcagacc gtcctcaacc agctcttccc
     3301 cggctacggc aatggccggc tcgaggacta caaagaccat gacggtgatt ataaagatca
     3361 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc
     3421 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3481 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3541 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3601 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3661 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3721 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3781 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3841 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3901 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3961 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4021 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4081 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4141 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4201 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4261 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4321 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4381 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4441 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4501 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4561 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4621 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4681 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4741 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4801 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4861 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4921 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4981 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5041 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5101 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5161 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5221 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5281 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5341 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5401 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5461 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5521 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5581 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5641 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5701 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5761 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5821 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5881 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6601 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6661 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6721 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6781 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6841 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6901 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6961 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7021 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7081 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7141 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7201 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7261 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7321 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7381 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7441 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7501 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7561 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7621 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7681 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7801 catttccccg aaaagtgcca cctgacgtc
//
