LOCUS       V038877                 8544 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038877
VERSION     V038877
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             756..3224
                     /label="USP16(NM_006447)"
                     /note="USP16(NM_006447)"
                     /gene="USP16"
     CDS             3231..3296
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3320..3908
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3974..4022
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4224..4652
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4666..4995
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5062..5856
                     /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    6032..6153
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6202..6218)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6226..6242
                     /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(6250..6280)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6295..6316
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6604..7192)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7363..8223)
                     /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(8224..8328)
                     /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 tggtaccgag ctcgaattcg ccaccatggg aaagaaacgg acaaagggaa
      781 aaactgttcc aatcgatgat tcctctgaaa ctttagaacc tgtgtgcaga cacattagaa
      841 aaggattgga acaaggtaat ttgaaaaagg ctttagtgaa tgtggaatgg aatatctgcc
      901 aagactgtaa gactgacaat aaagtgaaag ataaagctga agaagaaaca gaagaaaagc
      961 cttcagtttg gctgtgtctt aaatgtggcc atcagggctg tggcagaaat tctcaggagc
     1021 agcatgcctt gaagcactat ctgacgccaa gatctgaacc tcactgtctg gttcttagtt
     1081 tggacaactg gagtgtatgg tgttacgtat gtgataatga ggtccagtat tgtagttcaa
     1141 accagttggg tcaagtggtt gattatgtca gaaaacaagc cagcattaca actccaaagc
     1201 cagcagagaa agataatgga aatattgaac ttgaaaataa aaaattagaa aaagagagta
     1261 agaatgaaca agagagagaa aagaaggaaa acatggctaa agagaatcct cccatgaatt
     1321 ctccttgcca aataaccgtg aaaggactca gtaatttggg aaacacatgt ttcttcaatg
     1381 cagttatgca gaacttgtca caaacaccag tgcttagaga actactaaaa gaagtgaaaa
     1441 tgtctggaac aattgtaaaa attgaaccac ctgatttggc attaacagaa ccattagaaa
     1501 taaaccttga gcctccaggc cctcttactt tagccatgag ccagtttctt aatgagatgc
     1561 aagagaccaa aaagggggtt gtgacaccga aagaactctt ttctcaggtc tgtaaaaaag
     1621 cagtgcggtt taaaggctat cagcagcaag acagccagga gctgcttcgc tacttattgg
     1681 atgggatgag agcagaagaa caccaaagag tgagtaaagg aatacttaaa gcatttggta
     1741 attctactga aaagttggat gaagaactaa aaaataaagt taaagattat gagaagaaaa
     1801 aatcaatgcc aagttttgtt gaccgcatct ttggtggtga actaactagt atgatcatgt
     1861 gtgatcaatg cagaactgtc tccttggttc atgaatcttt ccttgatttg tccctcccag
     1921 ttttagatga tcagagtggt aagaaaagtg taaatgataa aaatctgaaa aagacagtgg
     1981 aggatgaaga tcaagatagt gaggaagaaa aagataacga cagttacata aaagagagaa
     2041 gtgatattcc ttctggaaca agtaagcact tacagaaaaa agcaaagaaa caagccaaaa
     2101 agcaagccaa gaaccaacga agacaacaaa aaattcaagg aaaagttctt catttaaatg
     2161 atatttgtac tattgaccat cctgaagaca gtgaatatga agctgaaatg tcacttcaag
     2221 gagaagtaaa tattaaatcc aaccatattt cacaagaggg tgttatgcat aaagaatatt
     2281 gtgtcaacca gaaagatttg aatggccaag caaaaatgat cgaaagtgta actgacaatc
     2341 aaaaatccac agaggaagta gatatgaaaa atatcaacat ggataatgat ctggaggttt
     2401 taacatcttc tcccactagg aatttaaatg gtgcctacct aacggaaggg agcaatggag
     2461 aagtggacat ttccaatggt ttcaaaaacc taaatttgaa tgctgctctt catcctgatg
     2521 aaataaatat agagattctg aatgatagtc atactcctgg aacaaaggtg tatgaggttg
     2581 taaatgaaga tccagaaact gctttctgta ctcttgcaaa cagggaagtt ttcaatactg
     2641 atgagtgttc aatccaacat tgtttatatc agttcacccg taatgagaaa cttcgagatg
     2701 cgaataaact gctttgtgaa gtatgcacac ggagacagtg taatggacca aaggcaaata
     2761 taaaaggtga aaggaagcat gtttacacca atgccaaaaa gcagatgcta atttctcttg
     2821 ctcctcctgt tcttactctt catttaaaga gatttcagca ggctggtttt aacctacgca
     2881 aagttaacaa acacataaag tttccggaaa tcttagattt ggctcctttt tgcaccctta
     2941 aatgtaagaa tgttgcagaa gaaaatacaa gggtactcta ttccttatat ggagttgttg
     3001 aacacagtgg tactatgagg tcggggcatt acactgccta tgccaaggca agaaccgcaa
     3061 atagtcatct ctctaatctt gttcttcacg gtgatattcc acaagatttt gaaatggaat
     3121 caaaagggca gtggtttcac atcagcgaca cacatgtgca agctgtgcct acaactaaag
     3181 tactaaactc acaagcgtac ctcctatttt atgagagaat actgggatcc gactacaaag
     3241 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     3301 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3361 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3421 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3481 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3541 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3601 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3661 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3721 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3781 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3841 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3901 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     3961 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     4021 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4081 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4141 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     4201 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4261 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4321 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4381 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4441 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4501 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4561 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     4621 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4681 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4741 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4801 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4861 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4921 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4981 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5041 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5101 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5161 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5221 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5281 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5341 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5401 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5461 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5521 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5581 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5641 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5701 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5761 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5821 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     5881 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5941 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6001 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6061 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6121 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6181 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6241 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6301 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6361 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6781 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7021 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7261 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7321 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7441 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7561 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7801 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7861 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8041 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8221 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8281 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8341 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     8401 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     8461 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     8521 aattgcatga agaatctgct tagg
//
