LOCUS       V036130                 7641 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036130
VERSION     V036130
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             928..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             988..1014
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1048..2442
                     /label="PRKN(NM_004562)"
                     /note="PRKN(NM_004562)"
                     /gene="PRKN"
     CDS             2479..3174
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="red fluorescent protein from Entacmaea
                     quadricolor"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR*"
                     /label="TurboRFP"
     polyA_signal    3241..3465
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3511..3939
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3953..4282
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4349..5143
                     /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    5317..5438
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5487..5503)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5511..5527
                     /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(5535..5565)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5580..5601
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5889..6474)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6645..7505)
                     /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(7506..7610)
                     /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 ggcggcgtac ccatacgacg tacctgatta tgcaacttat
      961 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatcagcg
     1021 gaaaagcttg gtaccgagct cggatccatg atagtgtttg tcaggttcaa ctccagccat
     1081 ggtttcccag tggaggtcga ttctgacacc agcatcttcc agctcaagga ggtggttgct
     1141 aagcgacagg gggttccggc tgaccagttg cgtgtgattt tcgcagggaa ggagctgagg
     1201 aatgactgga ctgtgcagaa ttgtgacctg gatcagcaga gcattgttca cattgtgcag
     1261 agaccgtgga gaaaaggtca agaaatgaat gcaactggag gcgacgaccc cagaaacgcg
     1321 gcgggaggct gtgagcggga gccccagagc ttgactcggg tggacctcag cagctcagtc
     1381 ctcccaggag actctgtggg gctggctgtc attctgcaca ctgacagcag gaaggactca
     1441 ccaccagctg gaagtccagc aggtagatca atctacaaca gcttttatgt gtattgcaaa
     1501 ggcccctgtc aaagagtgca gccgggaaaa ctcagggtac agtgcagcac ctgcaggcag
     1561 gcaacgctca ccttgaccca gggtccatct tgctgggatg atgttttaat tccaaaccgg
     1621 atgagtggtg aatgccaatc cccacactgc cctgggacta gtgcagaatt tttctttaaa
     1681 tgtggagcac accccacctc tgacaaggaa acatcagtag ctttgcacct gatcgcaaca
     1741 aatagtcgga acatcacttg cattacgtgc acagacgtca ggagccccgt cctggttttc
     1801 cagtgcaact cccgccacgt gatttgctta gactgtttcc acttatactg tgtgacaaga
     1861 ctcaatgatc ggcagtttgt tcacgaccct caacttggct actccctgcc ttgtgtggct
     1921 ggctgtccca actccttgat taaagagctc catcacttca ggattctggg agaagagcag
     1981 tacaaccggt accagcagta tggtgcagag gagtgtgtcc tgcagatggg gggcgtgtta
     2041 tgcccccgcc ctggctgtgg agcggggctg ctgccggagc ctgaccagag gaaagtcacc
     2101 tgcgaagggg gcaatggcct gggctgtggg tttgccttct gccgggaatg taaagaagcg
     2161 taccatgaag gggagtgcag tgccgtattt gaagcctcag gaacaactac tcaggcctac
     2221 agagtcgatg aaagagccgc cgagcaggct cgttgggaag cagcctccaa agaaaccatc
     2281 aagaaaacca ccaagccctg tccccgctgc catgtaccag tggaaaaaaa tggaggctgc
     2341 atgcacatga agtgtccgca gccccagtgc aggctcgagt ggtgctggaa ctgtggctgc
     2401 gagtggaacc gcgtctgcat gggggaccac tggttcgacg tggaattcgg tggaggcgga
     2461 tcaggaggcg gtggatctat gagcgagctg atcaaggaga acatgcacat gaagctgtac
     2521 atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc
     2581 tacgagggca cccagaccat gaagatcaag gtggtcgagg gcggccctct ccccttcgcc
     2641 ttcgacatcc tggctaccag cttcatgtac ggcagcaaag ccttcatcaa ccacacccag
     2701 ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga gagaatcacc
     2761 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcttcca gaacggctgc
     2821 atcatctaca acgtcaagat caacggggtg aacttcccat ccaacggccc tgtgatgcag
     2881 aagaaaacac gcggctggga ggccaacacc gagatgctgt accccgctga cggcggcctg
     2941 agaggccaca gccagatggc cctgaagctc gtgggcgggg gctacctgca ctgctccttc
     3001 aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg cttccacttc
     3061 gtggaccaca gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac
     3121 gagatggctg tggccaagta ctgcgacctc cctagcaaac tggggcacag ataagatatc
     3181 cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     3241 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3301 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3361 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3421 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3481 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3541 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3601 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3661 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3721 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3781 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3841 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3901 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3961 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4021 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4081 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4141 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4201 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4261 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4321 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4381 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4441 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4501 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4561 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4621 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4681 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4741 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4801 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4861 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4921 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4981 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5041 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5101 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5161 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5221 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5281 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5341 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5401 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5461 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5521 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5581 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5641 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5701 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5761 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5821 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5881 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5941 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6001 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6061 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6121 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6181 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6241 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6301 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6361 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6421 agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6481 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6541 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6601 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6661 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6721 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6781 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6841 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6901 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6961 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7021 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7081 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7141 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7201 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7261 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7321 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7381 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7441 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7501 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7561 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7621 cgaaaagtgc cacctgacgt c
//
