LOCUS       V019361                 7043 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019361
VERSION     V019361
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..1723
                     /label="Zdhhc19(NM_199309)"
                     /note="Zdhhc19(NM_199309)"
                     /gene="Zdhhc19"
     CDS             1730..1795
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1819..2407
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2473..2521
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2723..3151
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3165..3494
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3561..4355
                     /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    4531..4652
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4701..4717)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4725..4741
                     /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(4749..4779)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4794..4815
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5103..5691)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5862..6722)
                     /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(6723..6827)
                     /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 ccatgccttt cttaaaggat gctgtgacac ttgtgaagga
      721 accacaacag ctgccctcca tccctctttc atggttcccc tcgagcgtgt ttgctgcctt
      781 caatgtaacg ctgctgctgt ttttgagtgg ccttttcttc ggattccctt gtaggtggct
      841 ggttcagaac ggggagtggg cctttcctgc catcacagga ccactcttca tcctcacctt
      901 cttcagtctc gtctcgctca acttctcaga ccctggtatc ttacatcgag gctccaccaa
      961 agaggacccc atgacggtgc atgttgttcg agtgaaccaa agggctttcc gcctggaatg
     1021 gtgccccaag tgcctcttcc atcgcccacc ccgaacctac cactgcccgt ggtgcaacat
     1081 ttgcgtggag gacttcgacc accattgcaa gtgggtcaat aactgcatcg gtcaccgcaa
     1141 cttccgcctc ttcatgctgc tggtcctgtc actctgtctc tactcgggag ccctgctggt
     1201 cacctgcctg acattcctat ttcgcacaag gcatctgccc ttctccctgg acaaggggat
     1261 ggccatcctg gtggctgtgc ccgctgcggg ctttttgatt ccgctcttcc tgctgctgct
     1321 gatccaggcc ctatctgtga gcagggcgga gagctcctac gagagcaagt gcaggtacca
     1381 tccggaatac aacccttttg accagggctt tgccaagaac tggtatctgg caatgtttgc
     1441 accactgggt cccaattaca tgtctgaagt tgtctgtctg caaagaccag tggggacagc
     1501 atggatccaa gagaagacaa agccctcacc accacgtcgt cctaaacact gtaggccagg
     1561 tcctccgggg ccccagcacc aacctcggcg cgttccaggg aaggggcccc cagggagtgg
     1621 agaggctgca gctctccaag agatgcgaag gctgcctgca tctgtggaaa agtccccagg
     1681 aggtccacgc caacccacgg ctgaacctgc cgctggagac cccctcgagg actacaaaga
     1741 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     1801 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1861 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1921 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     1981 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2041 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2101 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2161 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     2221 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2281 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2341 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2401 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     2461 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     2521 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     2581 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     2641 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     2701 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2761 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2821 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     2881 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2941 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3001 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3061 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     3121 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3241 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3301 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3361 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3421 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3481 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3541 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3601 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3661 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3721 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3781 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3841 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3901 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3961 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4021 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4081 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4141 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4201 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4261 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4321 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4381 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4441 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4501 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4561 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4621 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4681 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4741 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4801 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4861 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4921 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4981 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5041 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5281 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5521 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6001 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6601 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6721 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6841 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     6901 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     6961 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     7021 attgcatgaa gaatctgctt agg
//
