LOCUS       V021489                 7430 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021489
VERSION     V021489
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..2077
                     /label="PRKN(NM_004562)"
                     /note="PRKN(NM_004562)"
                     /gene="PRKN"
     CDS             2117..2182
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2206..2794
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2860..2908
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3110..3538
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3552..3881
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3948..4742
                     /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    4918..5039
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5088..5104)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5112..5128
                     /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(5136..5166)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5181..5202
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5490..6078)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6249..7109)
                     /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(7110..7214)
                     /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 ccatgatagt gtttgtcagg ttcaactcca gccatggttt
      721 cccagtggag gtcgattctg acaccagcat cttccagctc aaggaggtgg ttgctaagcg
      781 acagggggtt ccggctgacc agttgcgtgt gattttcgca gggaaggagc tgaggaatga
      841 ctggactgtg cagaattgtg acctggatca gcagagcatt gttcacattg tgcagagacc
      901 gtggagaaaa ggtcaagaaa tgaatgcaac tggaggcgac gaccccagaa acgcggcggg
      961 aggctgtgag cgggagcccc agagcttgac tcgggtggac ctcagcagct cagtcctccc
     1021 aggagactct gtggggctgg ctgtcattct gcacactgac agcaggaagg actcaccacc
     1081 agctggaagt ccagcaggta gatcaatcta caacagcttt tatgtgtatt gcaaaggccc
     1141 ctgtcaaaga gtgcagccgg gaaaactcag ggtacagtgc agcacctgca ggcaggcaac
     1201 gctcaccttg acccagggtc catcttgctg ggatgatgtt ttaattccaa accggatgag
     1261 tggtgaatgc caatccccac actgccctgg gactagtgca gaatttttct ttaaatgtgg
     1321 agcacacccc acctctgaca aggaaacatc agtagctttg cacctgatcg caacaaatag
     1381 tcggaacatc acttgcatta cgtgcacaga cgtcaggagc cccgtcctgg ttttccagtg
     1441 caactcccgc cacgtgattt gcttagactg tttccactta tactgtgtga caagactcaa
     1501 tgatcggcag tttgttcacg accctcaact tggctactcc ctgccttgtg tggctggctg
     1561 tcccaactcc ttgattaaag agctccatca cttcaggatt ctgggagaag agcagtacaa
     1621 ccggtaccag cagtatggtg cagaggagtg tgtcctgcag atggggggcg tgttatgccc
     1681 ccgccctggc tgtggagcgg ggctgctgcc ggagcctgac cagaggaaag tcacctgcga
     1741 agggggcaat ggcctgggct gtgggtttgc cttctgccgg gaatgtaaag aagcgtacca
     1801 tgaaggggag tgcagtgccg tatttgaagc ctcaggaaca actactcagg cctacagagt
     1861 cgatgaaaga gccgccgagc aggctcgttg ggaagcagcc tccaaagaaa ccatcaagaa
     1921 aaccaccaag ccctgtcccc gctgccatgt accagtggaa aaaaatggag gctgcatgca
     1981 catgaagtgt ccgcagcccc agtgcaggct cgagtggtgc tggaactgtg gctgcgagtg
     2041 gaaccgcgtc tgcatggggg accactggtt cgacgtggaa ttctgcagat atccagcaca
     2101 gtggcggccg ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2161 ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca acctctggat
     2221 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2281 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2341 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2401 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2461 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2521 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2581 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     2641 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2701 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2761 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     2821 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     2881 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     2941 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     3001 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     3061 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     3121 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3181 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3241 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     3301 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3361 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3421 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3481 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3541 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3721 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3781 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3841 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3901 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3961 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4021 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4081 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4141 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4201 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4261 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4321 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4381 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4441 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4501 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4561 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4621 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4681 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4741 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4801 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4861 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4921 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4981 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5041 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5101 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5161 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5221 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5281 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5341 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5401 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5461 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5521 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5581 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5641 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     5701 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5761 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5821 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5881 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5941 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6001 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6061 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6121 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6181 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6241 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6301 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6361 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6421 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6481 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6541 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6601 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6661 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6721 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6781 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6841 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6901 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6961 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7021 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7081 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7141 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7201 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7261 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     7321 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     7381 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
