LOCUS       V018499                 7135 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018499
VERSION     V018499
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      901..910
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             964..2688
                     /label="ZNF587(NM_032828)"
                     /note="ZNF587(NM_032828)"
                     /gene="ZNF587"
     polyA_signal    2735..2959
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3005..3433
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3447..3776
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3843..4637
                     /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    4811..4932
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4981..4997)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5005..5021
                     /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(5029..5059)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5074..5095
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5383..5968)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6139..6999)
                     /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(7000..7104)
                     /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 gccaccatgg attacaagga tgacgacgat aagactaagc ttatgtccag tgtggtggaa
      961 ttcatggcag cggctgtgcc gaggcgccca actcagcagg gcactgtgac ctttgaagat
     1021 gtggctgtga acttttccca ggaggagtgg tgtcttctta gtgaggctca gaggtgcttg
     1081 taccgtgatg tgatgctaga gaacctggct ctcatatcct cgctgggttg ttggtgtgga
     1141 tcaaaagatg aggaggcacc ttgtaagcag agaatttctg tacaaagaga gtctcagagc
     1201 aggactccta gggcaggtgt ttctcctaag aaggctcacc cctgtgaaat gtgtggcctc
     1261 atcttggagg atgtttttca ctttgctgac caccaggaaa ctcatcacaa gcagaagctg
     1321 aacaggagtg gagcatgtgg aaaaaacttg gatgacactg cataccttca tcagcaccag
     1381 aagcagcata ttggagagaa attctacaga aagagtgtca gagaagcatc gtttgtaaag
     1441 aaacgtaagc tcagggtgtc acaggagcca tttgtcttcc gcgagtttgg gaaggacgtt
     1501 ctgcccagtt caggattgtg ccaagaagaa gctgctgtag agaagacaga cagtgaaact
     1561 atgcatggcc caccctttca ggagggaaaa actaattaca gttgtggaaa acgcacaaaa
     1621 gccttcagca ccaaacactc agttattcca caccagaaac ttttcactag agatggatgt
     1681 tatgtgtgca gtgattgtgg aaaatccttt agcagatatg tcagcttcag taatcatcag
     1741 cgagatcaca ctgcaaaagg accttatgat tgtggagagt gtgggaaatc ttatagtcga
     1801 aagagcagcc ttattcaaca tcagcgagtc cacactggac agacagctta tccctgtgag
     1861 gagtgcggga aatcttttag tcagaagggc agccttatta gccatcagct tgttcacact
     1921 ggagaagggc cttatgagtg tagagaatgt gggaaatctt ttggtcaaaa gggtaacctc
     1981 attcaacatc agcaaggtca cactggagag agagcttatc actgtgggga atgtgggaaa
     2041 tcttttcgtc agaagttctg ctttattaac catcagcgtg ttcacactgg agaaaggcct
     2101 tacaagtgtg gagaatgtgg gaaatctttt ggtcaaaagg gcaacctcgt tcaccatcag
     2161 cgaggtcata ctggagaaag gccctatgag tgcaaggaat gtgggaaatc atttaggtac
     2221 agatcccacc tcactgaaca ccagagactt cacactgggg aaagacctta caattgtagg
     2281 gaatgtggga aattatttaa caggaagtat catcttctgg ttcatgagag agttcacact
     2341 ggagaaaggc catatgcgtg tgaggtatgt gggaaattat ttggcaataa gcacagcgtg
     2401 actatacatc agaggattca cactggagaa aggccgtatg aatgcagtga atgtgggaaa
     2461 tcatttcttt ccagctctgc gcttcatgtt cataaaagag ttcattctgg acaaaagcct
     2521 tataagtgca gtgaatgtgg aaaatccttt tctgaatgtt ccagtctcat taaacacagg
     2581 agaattcaca ctggagaaag gccttatgaa tgcaccaaat gtggaaaaac atttcagcga
     2641 agctctaccc tccttcatca tcagagttca cacaggagaa aggccttatg actcgagtct
     2701 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2761 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2821 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2881 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2941 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3001 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3061 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3121 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3181 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3241 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3301 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3361 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3421 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3481 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3541 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3601 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3661 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3721 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3781 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3841 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3901 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3961 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4021 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4081 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4141 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4201 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4261 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4321 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4381 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4441 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4501 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4561 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4621 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4681 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4741 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4801 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4861 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4921 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4981 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5041 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5101 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5161 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5221 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5281 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5821 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     5941 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6001 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6061 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6121 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6181 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6241 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6301 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6361 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6421 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6481 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6541 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6601 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6661 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6721 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6781 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6841 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6901 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6961 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7021 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7081 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
