LOCUS       V019347                 7454 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019347
VERSION     V019347
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..2134
                     /label="Zdhhc1(NM_001379007)"
                     /note="Zdhhc1(NM_001379007)"
                     /gene="Zdhhc1"
     CDS             2141..2206
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2230..2818
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2884..2932
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3134..3562
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3576..3905
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3972..4766
                     /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    4942..5063
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5112..5128)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5136..5152
                     /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(5160..5190)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5205..5226
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5514..6102)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6273..7133)
                     /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(7134..7238)
                     /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 ccatgaacat ctgcaacaaa ccctccaaca agacagcccc
      721 cgagaagagt gtgtggacag cgccctcaca ggacagtggg ccttccccag aactgcaggg
      781 ccagcgctct cgaaggaatg gatggagctg gccgccccac ccgctccaga ttgtggcctg
      841 gctgctctac ctcttcttcg cggtgattgg ctttggggtc cttgttcccc tcttgcctca
      901 ccactgggtg cctgctggct atgcttgcat gggggccatc tttgctggcc accttgtggt
      961 gcacctgact gctgtctcca tcgatccagc agatgccaat gtgcgggaca agagctactc
     1021 tgggcccctg cccatcttca accgcagcca gcatgcgcat gtcattgaag acctgcactg
     1081 caacctatgc gacgtggacg tgagtgcgcg atccaaacac tgcagcgcct gcaataagtg
     1141 cgtatgcggc ttcgatcacc attgcaagtg gctcaacaac tgcgtgggcg agaggaacta
     1201 tcggctcttt ctacacagtg tggcatctgc tttattgggt gtcctgctcc tggtgctggt
     1261 ggccacttat gtcttcgtgg agttctttgt caaccccatg cgacttcgta ccaaccaaca
     1321 ctttgaagtc ttgaagaacc acacggatgt gtggtttgtg ttcttgcctg ctgcccctgt
     1381 ggagacccag gctcctgcca tcctggccct ggccgccctg ctcatccttc tgggcctgct
     1441 ttctacagcc ctgcttggcc acctgctctg cttccacatt tatctaatgt ggcacaagct
     1501 caccacctat gaatacatcg tgcagcaccg tccagctcag gaagcaaagg agacccacaa
     1561 ggagctggag tcatgccccc gcaaggtgcg gtccattcag gagatggagt tctacatgag
     1621 gaccttcagc catgtgcgtc cagagccctc tggccaggct aggacagcag cattaaatgc
     1681 caatccttcc cagtttcttg ccacccaagg ccaagtggaa ccaccactac cctcctcctc
     1741 agacacactg gctctgcctc cacggatcca accccagaaa aagaggaagc ggcgagtgta
     1801 tagattgccc aggtctgggg tcttggaccg agagctccca ctgcccaggt tacgggagac
     1861 tgggactcct agccgccgct ccagctcttc atctgattcc accagtgcca gcccagtgca
     1921 tgctggtggc tctgcaggcg cctactactc ggcatcagct gagtccatgg aagagattcc
     1981 agtggcccag acgcgcctgg gcagtgccgc actgggtgcc ccaggagcca ggggtcgaga
     2041 gtctgggctg gctctacagg cacgttcgcc tgctgttttt gtgagcccga gcagtggcga
     2101 gcccggaaca cctggaggcg gcgacggcct gcctctcgag gactacaaag accatgacgg
     2161 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     2221 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2281 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2341 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2401 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2461 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2521 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2581 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2641 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2701 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2761 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2821 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     2881 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     2941 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3001 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3061 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3121 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3181 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3241 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3301 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3361 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3421 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3481 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     3541 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3601 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3721 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3781 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3841 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3901 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3961 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4021 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4081 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4141 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4201 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4261 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4321 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4381 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4441 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4501 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4561 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4621 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4681 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4741 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4801 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4861 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4921 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4981 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5041 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5101 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5161 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5221 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5281 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5341 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5401 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5461 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5521 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5581 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5641 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5701 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5761 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5821 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5881 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5941 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6001 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6061 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6121 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6181 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6241 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6301 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6361 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6421 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6481 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6541 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6601 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6661 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6721 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6781 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6841 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6901 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6961 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7021 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7081 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7141 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7201 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7261 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7321 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7381 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7441 agaatctgct tagg
//
