LOCUS       V030645                 7574 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030645
VERSION     V030645
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..2233
                     /label="USP30(NM_032663)"
                     /note="USP30(NM_032663)"
                     /gene="USP30"
     CDS             2240..2266
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2270..2296
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2300..2326
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2350..2938
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3004..3052
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3254..3682
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3696..4025
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4092..4886
                     /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    5062..5183
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5232..5248)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5256..5272
                     /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(5280..5310)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5325..5346
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5634..6222)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6393..7253)
                     /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(7254..7358)
                     /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 ccatgctgag ctcccgggcc gaggcggcga tgaccgcggc
      721 cgacagggcc atccagcgct tcctgcggac cggggcggcc gtcagatata aagtcatgaa
      781 gaactgggga gttataggtg gaattgctgc tgctcttgca gcaggaatat atgttatttg
      841 gggtcccatt acagaaagaa agaagcgtag aaaagggctt gtgcctggcc ttgttaattt
      901 agggaacacc tgcttcatga actccctgct acaaggcctg tctgcctgtc ctgctttcat
      961 caggtggctg gaagagttca cctcccagta ctccagggat cagaaggagc ccccctcaca
     1021 ccagtattta tccttaacac tcttgcacct tctgaaagcc ttgtcctgcc aagaagttac
     1081 tgatgatgag gtcttagatg caagctgctt gttggatgtc ttaagaatgt acagatggca
     1141 gatctcatca tttgaagaac aggatgctca cgaattattc catgtcatta cctcgtcatt
     1201 ggaagatgag cgagaccgcc agcctcgggt cacacatttg tttgatgtgc attccctgga
     1261 gcagcagtca gaaataactc ccaaacaaat tacctgccgc acaagagggt cacctcaccc
     1321 tacatccaat cactggaagt ctcaacatcc ttttcatgga agactcacta gtaatatggt
     1381 ctgcaaacac tgtgaacacc agagtcctgt tcgatttgat acctttgata gcctttcact
     1441 aagtattcca gccgccacat ggggtcaccc attgaccctg gaccactgcc ttcaccactt
     1501 catctcatca gaatcagtgc gggatgttgt gtgtgacaac tgtacaaaga ttgaagccaa
     1561 gggaacgttg aacggggaaa aggtggaaca ccagaggacc acttttgtta aacagttaaa
     1621 actagggaag ctccctcagt gtctctgcat ccacctacag cggctgagct ggtccagcca
     1681 cggcacgcct ctgaagcggc atgagcacgt gcagttcaat gagttcctga tgatggacat
     1741 ttacaagtac cacctccttg gacataaacc tagtcaacac aaccctaaac tgaacaagaa
     1801 cccagggcct acactggagc tgcaggatgg gccgggagcc cccacaccag ttctgaatca
     1861 gccaggggcc cccaaaacac agatttttat gaatggcgcc tgctccccat ctttattgcc
     1921 aacgctgtca gcgccgatgc ccttccctct cccagttgtt cccgactaca gctcctccac
     1981 atacctcttc cggctgatgg cagttgtcgt ccaccatgga gacatgcact ctggacactt
     2041 tgtcacttac cgacggtccc caccttctgc caggaaccct ctctcaacta gcaatcagtg
     2101 gctgtgggtc tccgatgaca ctgtccgcaa ggccagcctg caggaggtcc tgtcctccag
     2161 cgcctacctg ctgttctacg agcgcgtcct ttccaggatg cagcaccaga gccaggagtg
     2221 caagtctgaa gaactcgagt acccatacga cgtacctgat tatgcaactt atccttatga
     2281 tgtacctgat tatgctagct atccttatga tgtcccagac tacgcatagt gagggcccga
     2341 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2401 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2461 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2521 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2581 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2641 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2701 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2761 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2821 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2881 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2941 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3001 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     3061 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3121 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3181 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3241 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3301 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3361 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3421 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3481 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3541 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3601 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     3661 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3721 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3841 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3901 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3961 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4021 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4081 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4141 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4201 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4261 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4321 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4381 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4441 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4501 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4561 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4621 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4681 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4741 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4801 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4861 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4921 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4981 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5041 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5101 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5161 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5221 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5281 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5341 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5401 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5461 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5821 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6061 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7381 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7441 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7501 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7561 agaatctgct tagg
//
