LOCUS       V023579                 7141 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023579
VERSION     V023579
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"
     regulatory      655..664
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             664..729
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             751..1851
                     /label="ZDHHC2(NM_016353)"
                     /note="ZDHHC2(NM_016353)"
                     /gene="ZDHHC2"
     misc_feature    1917..2505
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2571..2619
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2821..3249
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3263..3592
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3659..4453
                     /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    4629..4750
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4799..4815)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4823..4839
                     /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(4847..4877)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4892..4913
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5201..5789)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5960..6820)
                     /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(6821..6925)
                     /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 ttaagccacc
      661 atggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
      721 gatgacaagc ttggtaccga gctcggatcc atggcgccct cgggcccggg cagcagcgcc
      781 aggcggcggt gccggcgggt gctgtactgg atcccggtgg tgttcatcac cctcctgctc
      841 ggctggtcct actacgccta cgccatccag ctgtgcatag tgtccatgga aaacactggc
      901 gaacaagttg tgtgcctgat ggcctatcat ctactttttg caatgtttgt ctggtcatac
      961 tggaaaacta tctttacatt accaatgaat ccttcaaaag aattccatct ctcttatgca
     1021 gagaaagatt tgttggagag agagccaaga ggagaagccc atcaggaagt tcttaggcga
     1081 gcagccaagg atcttcccat ctataccagg accatgtctg gagccatccg atactgtgac
     1141 agatgccaac ttataaaacc agatcgctgc catcactgct ccgtctgtga taaatgtatt
     1201 ttgaagatgg atcatcattg tccatgggtg aacaattgtg ttggattttc aaattataag
     1261 ttctttctcc ttttcttggc ttattctctg ctctactgcc tttttattgc ggcaacagat
     1321 ttacagtatt ttatcaaatt ttggacaaat ggcctacctg atactcaagc caagttccat
     1381 attatgtttt tattctttgc tgcagctatg ttttctgtca gcttgtcttc tctgtttggc
     1441 tatcattgtt ggctagtcag caaaaataaa tctacattag aggcattcag aagtccagta
     1501 tttcgacatg gaacagataa gaatggattc agcttgggtt tcagtaaaaa catgcgacaa
     1561 gtttttggtg atgagaagaa gtactggttg ctacccattt tttcaagtct aggtgatggc
     1621 tgctcctttc caacttgcct tgttaaccag gatcctgaac aagcatctac tcctgcaggg
     1681 ctgaattcca cagctaaaaa tctcgaaaac catcagtttc ctgcaaagcc attgagagag
     1741 tcccagagcc accttcttac tgattctcag tcttggacgg agagcagcat aaacccagga
     1801 aaatgcaaag ctggtatgag caatcctgca ttaaccatgg aaaatgagac ttaagaattc
     1861 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgatat ctcgacaatc
     1921 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1981 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2041 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2101 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2161 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2221 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2281 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     2341 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2401 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2461 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag
     2521 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     2581 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     2641 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     2701 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     2761 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     2821 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2881 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2941 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     3001 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3061 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3121 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3181 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3241 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3301 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3361 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3421 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3481 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3541 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3601 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3661 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3721 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3781 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3841 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3901 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3961 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4021 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4081 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4141 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4201 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4261 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4321 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4381 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4441 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     4501 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4561 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4621 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4681 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4741 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4801 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4861 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4921 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4981 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5041 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5101 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5161 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5221 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5281 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5341 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     5401 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5461 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5521 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5581 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5641 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5701 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5761 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5821 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5881 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5941 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6001 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6061 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6121 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6181 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6241 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6301 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6361 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6421 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6481 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6541 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6601 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6661 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6721 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6781 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6841 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6901 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6961 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
     7021 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     7081 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     7141 g
//
