LOCUS       V031453                 8584 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031453
VERSION     V031453
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..4137
                     /label="OTUD4(NM_001102653)"
                     /note="OTUD4(NM_001102653)"
                     /gene="OTUD4"
     polyA_signal    4184..4408
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4454..4882
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4896..5225
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5292..6086
                     /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    6260..6381
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6430..6446)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6454..6470
                     /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(6478..6508)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6523..6544
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6832..7417)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7588..8448)
                     /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(8449..8553)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggcctgta ttcactatct tcgagagaac
     1021 agagagaaat ttgaagcgtt tatagaagga tcatttgaag aatatttaaa gcgtttggaa
     1081 aatccacagg aatgggtagg acaagtggaa ataagtgccc tttctcttat gtacaggaaa
     1141 gattttataa tttatcggga accaaatgtt tctccttcac aagtaacaga aaataatttt
     1201 cctgaaaagg tgttactgtg tttttcaaat ggaaatcatt atgatattgt gtatcccata
     1261 aagtataaag aaagctctgc tatgtgtcag tctctccttt atgaattgct gtatgagaag
     1321 gtatttaaaa ctgatgttag taaaattgtg atggaactag acacgttgga agtagctgat
     1381 gaagataaca gtgaaatatc agattcagag gatgacagtt gcaagagtaa gactgctgct
     1441 gctgctgctg atgtgaatgg atttaaacct ttgtcaggca atgagcagct gaagaacaat
     1501 gggaactcta ctagcctgcc tttgtctaga aaggttctta agtcactcaa tcctgcagtc
     1561 tatagaaatg tggaatatga aatttggctg aagtctaaac aagctcagca aaaacgtgat
     1621 tattccattg ctgctggctt acaatatgaa gttggagaca aatgtcaagt taggttggat
     1681 cacaatggaa aatttttgaa tgcagatgtt caaggaattc attctgagaa tggaccagtt
     1741 ttggttgaag aactgggaaa gaagcacaca tcaaagaacc tcaaggcacc tcccccagaa
     1801 agctggaaca cagtgtcagg gaagaagatg aaaaaacctt ccacttctgg acaaaatttc
     1861 cattctgatg tggattacag agggccaaag aatccaagca agccaataaa agccccatca
     1921 gcactacctc ctcgactgca gcatccttca ggagtaagac aacatgcgtt ctctagtcat
     1981 tcttcagggt cacagtctca gaaattctcc agtgagcaca aaaatcttag ccggacacct
     2041 tcacagatca taagaaaacc tgatcgtgaa agagttgagg attttgatca cacaagtcga
     2101 gaatctaact atttcggcct ttccccagaa gagcgcagag agaagcaagc tatagaagaa
     2161 tcccgtttac tctatgagat tcagaacaga gatgaacagg ctttcccagc cctttccagc
     2221 tcatcagtca atcagtcagc ttctcagagt agcaatccat gtgtccagag aaaatcatca
     2281 catgtaggtg atagaaaagg aagcaggcgg agaatggata cagaagaacg aaaagacaaa
     2341 gactctattc atggacatag tcagttggat aaaagacccg aaccaagcac attggagaat
     2401 attactgatg ataaatatgc aacagtttca tcaccatcaa agtcaaagaa gttagagtgc
     2461 ccttctcctg cggaacaaaa gccagcagaa catgtgtctt tgtcaaatcc agctcccctt
     2521 ctagtttctc cagaggtaca tctaactcct gcggtgcctt ctttaccagc cactgtgcca
     2581 gcctggccaa gtgaacctac aacttttgga ccaacaggtg tccctgctcc aattcccgtt
     2641 ttgtcagtga cacagacttt gaccactgga cctgattcag ctgtatccca agctcattta
     2701 acaccctctc cagttcctgt gtcaatacag gcagttaacc agcccttgat gcctttgcct
     2761 cagacattga gcctttatca agacccactc tatcctgggt ttccttgtaa tgaaaaggga
     2821 gatcgagcca ttgtaccacc ttattcactg tgtcagactg gggaggacct acctaaagat
     2881 aagaatattc ttcgattctt cttcaatctt ggtgtgaagg catacagttg tcctatgtgg
     2941 gccccacatt cttacctgta ccctctgcac caggcctacc tggcagcctg caggatgtac
     3001 ccaaaggtcc ctgtccctgt ttatcctcat aatccctggt tccaagaggc tcctgctgct
     3061 cagaatgaaa gtgattgtac ctgtactgat gcccactttc ctatgcagac tgaggccagt
     3121 gttaatggtc aaatgccaca gccagagatt ggaccgccga cattttcttc acctctggtt
     3181 atccctccat ctcaggtgtc tgaaagtcat ggacaattgt cttaccaggc tgatcttgaa
     3241 tctgagaccc ctgggcagct tctgcatgct gattatgaag agtcactaag tggcaagaat
     3301 atgttccccc agccatcttt tggacccaat ccattcttag gcccagttcc tattgcacct
     3361 cctttctttc ctcatgtttg gtatgggtac ccttttcagg gattcataga aaatccagta
     3421 atgaggcaga atattgtcct gccctctgat gaaaaaggag aattggatct gtctctggaa
     3481 aatctggatc tgtctaaaga ttgtggttca gtttcaacag tagatgagtt tccagaagcc
     3541 aggggtgaac atgtacattc tctccctgaa gcaagtgtga gcagtaagcc ggacgaaggc
     3601 cggacagagc aatcttccca gacacgaaag gcagatacgg cattggcttc catccctcct
     3661 gtagcagagg gaaaggctca tcctcccact cagattctaa acagagagag agaaactgtg
     3721 cctgttgaac ttgaacctaa aaggaccatt caaagcctga aagaaaaaac agaaaaagta
     3781 aaagatccta agactgctgc tgatgtggtc agccctgggg ccaactctgt tgatagcaga
     3841 gtgcaaagac caaaagaaga gagttcagaa gatgaaaatg aagtgtctaa tattttgaga
     3901 agtggtagat ccaagcagtt ctataatcaa acttatggaa gcaggaagta caaaagtgat
     3961 tggggctatt ctggtagggg tggatatcaa catgtgagaa gtgaggagtc ctggaaagga
     4021 cagccaagca gaagtcggga tgaaggttat cagtaccatc gaaatgtcag agggcgacca
     4081 tttaggggag ataggaggag atcagggatg ggagatggcc ataggggaca gcacacttga
     4141 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     4201 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4261 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4321 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4381 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     4441 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4501 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4561 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4621 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4681 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4741 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4801 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4861 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4921 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4981 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5041 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5101 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5161 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5221 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     5281 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5341 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5401 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5461 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5521 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5581 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5641 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5701 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5761 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5821 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5881 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5941 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6001 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6061 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     6121 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6181 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6241 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6301 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6361 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6421 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6541 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6601 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6661 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6721 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6781 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7261 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     7381 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7441 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7501 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7561 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7621 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7681 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7741 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7801 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7861 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7921 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7981 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8041 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8101 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8161 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8221 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8281 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8341 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8401 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8461 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8521 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8581 cgtc
//
