LOCUS       V033456                 7914 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033456
VERSION     V033456
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        4..383
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        386..661
                     /label="chicken β-actin promoter"
     intron          662..1674
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     regulatory      1758..1767
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             1773..1796
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1797..1910
                     /codon_start=1
                     /note="selected from a peptide library; binds streptavidin
                     with nanomolar affinity (Keefe et al., 2001)"
                     /product="streptavidin-binding peptide"
                     /transl_table=1
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
                     /label="SBP"
     CDS             1941..2114
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /transl_table=1
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
                     /label="ProtA"
     CDS             2115..2288
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /transl_table=1
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
                     /label="ProtA"
     CDS             2376..2549
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /transl_table=1
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
                     /label="ProtA"
     CDS             2550..2723
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /transl_table=1
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
                     /label="ProtA"
     CDS             2754..2774
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             2775..2798
                     /codon_start=1
                     /product="recognition and cleavage site for human
                     rhinovirus 3C and PreScission proteases"
                     /transl_table=1
                     /translation="LEVLFQGP"
                     /label="HRV 3C site"
     CDS             2805..4811
                     /label="HDAC10(NM_032019)"
                     /note="HDAC10(NM_032019)"
                     /gene="HDAC10"
     polyA_signal    5002..5057
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(5417..5433)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5441..5457
                     /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(5465..5495)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5510..5531
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5590..5785
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    5791..5925
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(6164..6752)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6923..7783)
                     /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(7784..7888)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattctg cagtcgacct
     1741 cgatcgacgg tatcgatgcc accatggcta gcgattacaa ggatgacgat gacaagatgg
     1801 acgagaagac caccggctgg cggggcggcc acgtggtgga gggcctggcc ggcgagctgg
     1861 agcagctgcg ggccaggctg gagcaccacc ctcagggcca gcgggagccc accgcggctc
     1921 ttgcgcaaca cgatgaagcc gtggacaaca aattcaacaa agaacaacaa aacgcgttct
     1981 atgagatctt acatttacct aacttaaacg aagaacaacg aaacgccttc atccaaagtt
     2041 taaaagatga cccaagccaa agcgctaacc ttttagcaga agctaaaaag ctaaatgatg
     2101 ctcaggcgcc gaaagtagac aacaaattca acaaagaaca acaaaacgcg ttctatgaga
     2161 tcttacattt acctaactta aacgaagaac aacgaaacgc cttcatccaa agtttaaaag
     2221 atgacccaag ccaaagcgct aaccttttag cagaagctaa aaagctaaat ggtgctcagg
     2281 cgccgaaagt agacgcgaat tccgcgggga agtcaaccgg agggtctggt gggagtgggg
     2341 gctcaaccgc ggctcttgcg caacacgatg aagccgtgga caacaaattc aacaaagaac
     2401 aacaaaacgc gttctatgag atcttacatt tacctaactt aaacgaagaa caacgaaacg
     2461 ccttcatcca aagtttaaaa gatgacccaa gccaaagcgc taacctttta gcagaagcta
     2521 aaaagctaaa tgatgctcag gcgccgaaag tagacaacaa attcaacaaa gaacaacaaa
     2581 acgcgttcta tgagatctta catttaccta acttaaacga agaacaacga aacgccttca
     2641 tccaaagttt aaaagatgac ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc
     2701 taaatggtgc tcaggcgccg aaagtagacg cgaattccgc ggggaagtca accgagaacc
     2761 tgtacttcca atccctggaa gttctgttcc aggggcccga gctcatgggg accgcgcttg
     2821 tgtaccatga ggacatgacg gccacccggc tgctctggga cgaccccgag tgcgagatcg
     2881 agcgtcctga gcgcctgacc gcagccctgg atcgcctgcg gcagcgcggc ctggaacaga
     2941 ggtgtctgcg gttgtcagcc cgcgaggcct cggaagagga gctgggcctg gtgcacagcc
     3001 cagagtatgt atccctggtc agggagaccc aggtcctagg caaggaggag ctgcaggcgc
     3061 tgtccggaca gttcgacgcc atctacttcc acccgagtac ctttcactgc gcgcggctgg
     3121 ccgcaggggc tggactgcag ctggtggacg ctgtgctcac tggagctgtg caaaatgggc
     3181 ttgccctggt gaggcctccc gggcaccatg gccagagggc ggctgccaac gggttctgtg
     3241 tgttcaacaa cgtggccata gcagctgcac atgccaagca gaaacacggg ctacacagga
     3301 tcctcgtcgt ggactgggat gtgcaccatg gccaggggat ccagtatctc tttgaggatg
     3361 accccagcgt cctttacttc tcctggcacc gctatgagca tgggcgcttc tggcctttcc
     3421 tgcgagagtc agatgcagac gcagtggggc ggggacaggg cctcggcttc actgtcaacc
     3481 tgccctggaa ccaggttggg atgggaaacg ctgactacgt ggctgccttc ctgcacctgc
     3541 tgctcccact ggcctttgag tttgaccctg agctggtgct ggtctcggca ggatttgact
     3601 cagccatcgg ggaccctgag gggcaaatgc aggccacgcc agagtgcttc gcccacctca
     3661 cacagctgct gcaggtgctg gccggcggcc gggtctgtgc cgtgctggag ggcggctacc
     3721 acctggagtc actggcggag tcagtgtgca tgacagtaca gacgctgctg ggtgacccgg
     3781 ccccacccct gtcagggcca atggcgccat gtcagagtgc cctagagtcc atccagagtg
     3841 cccgtgctgc ccaggccccg cactggaaga gcctccagca gcaagatgtg accgctgtgc
     3901 cgatgagccc cagcagccac tccccagagg ggaggcctcc acctctgctg cctgggggtc
     3961 cagtgtgtaa ggcagctgca tctgcaccga gctccctcct ggaccagccg tgcctctgcc
     4021 ccgcaccctc tgtccgcacc gctgttgccc tgacaacgcc ggatatcaca ttggttctgc
     4081 cccctgacgt catccaacag gaagcgtcag ccctgaggga ggagacagaa gcctgggcca
     4141 ggccacacga gtccctggcc cgggaggagg ccctcactgc acttgggaag ctcctgtacc
     4201 tcttagatgg gatgctggat gggcaggtga acagtggtat agcagccact ccagcctctg
     4261 ctgcagcagc caccctggat gtggctgttc ggagaggcct gtcccacgga gcccagaggc
     4321 tgctgtgcgt ggccctggga cagctggacc ggcctccaga cctcgcccat gacgggagga
     4381 gtctgtggct gaacatcagg ggcaaggagg cggctgccct atccatgttc catgtctcca
     4441 cgccactgcc agtgatgacc ggtggtttcc tgagctgcat cttgggcttg gtgctgcccc
     4501 tggcctatgg cttccagcct gacctggtgc tggtggcgct ggggcctggc catggcctgc
     4561 agggccccca cgctgcactc ctggctgcaa tgcttcgggg gctggcaggg ggccgagtcc
     4621 tggccctcct ggaggagaac tccacacccc agctagcagg gatcctggcc cgggtgctga
     4681 atggagaggc acctcctagc ctaggccctt cctctgtggc ctccccagag gacgtccagg
     4741 ccctgatgta cctgagaggg cagctggagc ctcagtggaa gatgttgcag tgccatcctc
     4801 acctggtggc ttgagcggcc gcacggctcg agcggacgcg tcggtaagtc gaggaattca
     4861 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     4921 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     4981 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     5041 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     5101 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     5161 gtggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     5221 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     5281 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     5341 actactccca gtcatagctg tccctcttct cttatgaaga tccctcgacc tgcagcccaa
     5401 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5461 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5521 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5581 agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5641 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5701 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5761 ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac
     5821 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5881 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa
     5941 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6001 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6061 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6121 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6181 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6241 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6301 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     6361 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6421 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6481 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6541 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6601 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6661 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6721 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6781 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6841 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6901 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6961 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7021 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7081 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7141 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7201 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7261 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7321 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7381 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7441 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7501 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7561 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7621 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7681 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7741 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7801 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7861 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctg
//
