LOCUS       V019415                 7868 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019415
VERSION     V019415
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..2548
                     /label="Zdhhc13(NM_028031)"
                     /note="Zdhhc13(NM_028031)"
                     /gene="Zdhhc13"
     CDS             2555..2620
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2644..3232
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3298..3346
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3548..3976
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3990..4319
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4386..5180
                     /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    5356..5477
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5526..5542)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5550..5566
                     /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(5574..5604)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5619..5640
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5928..6516)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6687..7547)
                     /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(7548..7652)
                     /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 ccatggaggg cccgggcctg ggctcgcagt gcaggaatca
      721 cagccatggc tcccacgttc caggatttgg gcgacacggc atctgtgtac atgaaaacaa
      781 agaacttgca aaagcaaaag aaattcttcc tcttatagag gactcgagta actgtgacat
      841 tgtcaaagct acacaatatg gaatttttga acgatgtaaa gagctggtag aagcaggata
      901 tgatgtcaga caaccagaca gagaaaatgt gtctcttctt cactgggctg ccattaacaa
      961 caggctggaa cttgtaaagt tttatatttc caaaggtgct gtgatagatc agttgggtgg
     1021 tgacctaaat tcaactcctc ttcactgggc catccgacaa gggcatttgc ctatggtcat
     1081 attattgctc cagcacggtg cagaccccac tctgattgat ggagagggat tcagcagcat
     1141 ccatctggcg gtcctgtttc agcacatgcc catcatagcg tatctcatct caaagggaca
     1201 gagtgtgaat atgacagatg tcaatggaca gacgccgctc atgctatcgg cttacaaagt
     1261 aattgggcca gaaccaactg gatttctttt aaagttcaat ccctctctca gtgttgttga
     1321 taaaacacat cagaacactc ctcttcactg ggcagttgca gcaggaaatg tcagtgcagt
     1381 tgataagctt ttggaagctg gttctagcct ggacatccga aatgctaagg gagaaacacc
     1441 tcttgatatg gctctacaaa gtaaaaacca gctcattagt cacatgctaa gaacagaggc
     1501 caagatgaga gccaacaaac agtttcggct ttggcgatgg ctgcacaaat gcgagctctt
     1561 cctgctgctg atactctcta tgatcaccct gtgggctgtt ggatacatcc tggacttcaa
     1621 ttcagattct tggcttttaa aaggatgtct tctagtagca ttgttttttc tgacatcttt
     1681 gtttccaagg tttttggttg gctataagaa ccttgtgtac ttaccgacag tctttctgct
     1741 aagttccatt ttttggatat ttatgacttg gttcatctta ttctttcctg atacagcagg
     1801 tagccctttg tattttgctt ttattttcag catcatggcc tttctctatt ttttctacaa
     1861 gacttgggca actgatccag gcttcactaa ggcttcggag gaagagagga aagtgaatat
     1921 cgtcacccta gcagagactg gctctctgga cttcagaaca ttttgcacat catgtcttat
     1981 aaggaagcct ttaagatcac tccattgcca cgtgtgcaac tcctgtgtgg ctcgctttga
     2041 tcagcactgc ttttggactg gacgctgcat aggttttggc aaccatcatc attacatctt
     2101 cttcttgctt tccctttcca tggtatgtga ctggatcata tatggatctt tcgtctactg
     2161 gtcaaatcat tgtgccacaa cattcaagga agatggacta tggacctacc tcaatcagat
     2221 agtggcctgt tctccctggg ttctgtatat cttcatgcta gcagctttcc acttctcatg
     2281 gtcaacattt ttattaataa atcaactctt tcagattgca tttctgggcc taacctccca
     2341 tgagagaatc agcctcttaa agcaaagcag gcacatgaaa cagacattgt ccctcaggaa
     2401 aacaccatac aacctcggat tcacacagaa cctggcagat ttctttcagt gtggctgctt
     2461 tggcttagtg aagccctgca tcatagattg gacatcccag tacaccatgg ttttccaccc
     2521 agccaaagag aaagtccttc gctctgtact cgaggactac aaagaccatg acggtgatta
     2581 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc
     2641 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2701 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2761 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2821 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2881 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2941 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3001 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     3061 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3121 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3181 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     3241 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3301 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     3361 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3421 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3481 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     3541 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3601 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3661 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     3721 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3781 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3841 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3901 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3961 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4021 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4081 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4141 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4201 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4261 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4321 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4381 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4441 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4501 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4561 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4621 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4681 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4741 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4801 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4861 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4921 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4981 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5041 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5101 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5161 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     5221 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5281 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5341 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5401 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5461 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5521 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5581 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5641 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5701 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6121 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6361 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6601 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6661 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6721 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6781 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6841 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6901 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6961 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7021 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7081 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7141 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7201 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7261 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7321 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7381 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7441 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7501 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7561 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7621 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7681 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     7741 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     7801 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     7861 tgcttagg
//
