LOCUS       V035366                 9789 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035366
VERSION     V035366
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             759..776
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             783..4469
                     /label="HDAC6(NM_001321225)"
                     /note="HDAC6(NM_001321225)"
                     /gene="HDAC6"
     CDS             4476..4541
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4565..5153
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5219..5267
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5469..5897
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5911..6240
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6307..7101
                     /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    7277..7398
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7447..7463)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7471..7487
                     /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(7495..7525)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7540..7561
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7849..8437)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8608..9468)
                     /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(9469..9573)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgca ccatcaccat caccatctcg
      781 agatgagtgg agcgaaccgc ggcaggggcc aagcctcctc aactatgacc tcaaccggcc
      841 aggattccac cacaaccagg cagcgaagaa gtaggcagaa cccccagtcg ccccctcagg
      901 actccagtgt cacttcgaag cgaaatatta aaaagggagc cgttccccgc tctatcccca
      961 atctagcgga ggtaaagaag aaaggcaaaa tgaagaagct cggccaagca atggaagaag
     1021 acctaatcgt gggactgcaa gggatggatc tgaaccttga ggctgaagca ctggctggca
     1081 ctggcttggt gttggatgag cagttaaatg aattccattg cctctgggat gacagcttcc
     1141 cggaaggccc tgagcggctc catgccatca aggagcaact gatccaggag ggcctcctag
     1201 atcgctgcgt gtcctttcag gcccggtttg ctgaaaagga agagctgatg ttggttcaca
     1261 gcctagaata tattgatctg atggaaacaa cccagtacat gaatgaggga gaactccgtg
     1321 tcctagcaga cacctacgac tcagtttatc tgcatccgaa ctcatactcc tgtgcctgcc
     1381 tggcctcagg ctctgtcctc aggctggtgg atgcggtcct gggggctgag atccggaatg
     1441 gcatggccat cattaggcct cctggacatc acgcccagca cagtcttatg gatggctatt
     1501 gcatgttcaa ccacgtggct gtggcagccc gctatgctca acagaaacac cgcatccgga
     1561 gggtccttat cgtagattgg gatgtgcacc acggtcaagg aacacagttc accttcgacc
     1621 aggaccccag tgtcctctat ttctccatcc accgctacga gcagggtagg ttctggcccc
     1681 acctgaaggc ctctaactgg tccaccacag gtttcggcca aggccaagga tataccatca
     1741 atgtgccttg gaaccaggtg gggatgcggg atgctgacta cattgctgct ttcctgcacg
     1801 tcctgctgcc agtcgccctc gagttccagc ctcagctggt cctggtggct gctggatttg
     1861 atgccctgca aggggacccc aagggtgaga tggccgccac tccggcaggg ttcgcccagc
     1921 taacccacct gctcatgggt ctggcaggag gcaagctgat cctgtctctg gagggtggct
     1981 acaacctccg cgccctggct gaaggcgtca gtgcttcgct ccacaccctt ctgggagacc
     2041 cttgccccat gctggagtca cctggtgccc cctgccggag tgcccaggct tcagtttcct
     2101 gtgctctgga agcccttgag cccttctggg aggttcttgt gagatcaact gagaccgtgg
     2161 agagggacaa catggaggag gacaatgtag aggagagcga ggaggaagga ccctgggagc
     2221 cccctgtgct cccaatcctg acatggccag tgctacagtc tcgcacaggg ctggtctatg
     2281 accaaaatat gatgaatcac tgcaacttgt gggacagcca ccaccctgag gtaccccagc
     2341 gcatcttgcg gatcatgtgc cgtctggagg agctgggcct tgccgggcgc tgcctcaccc
     2401 tgacaccgcg ccctgccaca gaggctgagc tgctcacctg tcacagtgct gagtacgtgg
     2461 gtcatctccg ggccacagag aaaatgaaaa cccgggagct gcaccgtgag agttccaact
     2521 ttgactccat ctatatctgc cccagtacct tcgcctgtgc acagcttgcc actggcgctg
     2581 cctgccgcct ggtggaggct gtgctctcag gagaggttct gaatggtgct gctgtggtgc
     2641 gtcccccagg acaccacgca gagcaggatg cagcttgcgg tttttgcttt ttcaactctg
     2701 tggctgtggc tgctcgccat gcccagacta tcagtgggca tgccctacgg atcctgattg
     2761 tggattggga tgtccaccac ggtaatggaa ctcagcacat gtttgaggat gaccccagtg
     2821 tgctatatgt gtccctgcac cgctatgatc atggcacctt cttccccatg ggggatgagg
     2881 gtgccagcag ccagatcggc cgggctgcgg gcacaggctt caccgtcaac gtggcatgga
     2941 acgggccccg catgggtgat gctgactacc tagctgcctg gcatcgcctg gtgcttccca
     3001 ttgcctacga gtttaaccca gaactggtgc tggtctcagc tggctttgat gctgcacggg
     3061 gggatccgct ggggggctgc caggtgtcac ctgagggtta tgcccacctc acccacctgc
     3121 tgatgggcct tgccagtggc cgcattatcc ttatcctaga gggtggctat aacctgacat
     3181 ccatctcaga gtccatggct gcctgcactc gctccctcct tggagaccca ccacccctgc
     3241 tgaccctgcc acggccccca ctatcagggg ccctggcctc aatcactgag accatccaag
     3301 tccatcgcag atactggcgc agcttacggg tcatgaaggt agaagacaga gaaggaccct
     3361 ccagttctaa gttggtcacc aagaaggcac cccaaccagc caaacctagg ttagctgagc
     3421 ggatgaccac acgagaaaag aaggttctgg aagcaggcat ggggaaagtc acctcggcat
     3481 catttgggga agagtccact ccaggccaga ctaactcaga gacagctgtg gtggccctca
     3541 ctcaggacca gccctcagag gcagccacag ggggagccac tctggcccag accatttctg
     3601 aggcagccat tgggggagcc atgctgggcc agaccacctc agaggaggct gtcgggggag
     3661 ccactccgga ccagaccacc tcagaggaga ctgtgggagg agccattctg gaccagacca
     3721 cctcagagga tgctgttggg ggagccacgc tgggccagac tacctcagag gaggctgtag
     3781 gaggagctac actggcccag accacctcgg aggcagccat ggagggagcc acactggacc
     3841 agactacgtc agaggaggct ccagggggca ccgagctgat ccaaactcct ctagcctcga
     3901 gcacagacca ccagaccccc ccaacctcac ctgtgcaggg aactacaccc cagatatctc
     3961 ccagtacact gattgggagt ctcaggacct tggagctagg cagcgaatct cagggggcct
     4021 cagaatctca ggccccagga gaggagaacc tactaggaga ggcagctgga ggtcaggaca
     4081 tggctgattc gatgctgatg cagggatcta ggggcctcac tgatcaggcc atattttatg
     4141 ctgtgacacc actgccctgg tgtccccatt tggtggcagt atgccccata cctgcagcag
     4201 gcctagacgt gacccaacct tgtggggact gtggaacaat ccaagagaat tgggtgtgtc
     4261 tctcttgcta tcaggtctac tgtggtcgtt acatcaatgg ccacatgctc caacaccatg
     4321 gaaattctgg acacccgctg gtcctcagct acatcgacct gtcagcctgg tgttactact
     4381 gtcaggccta tgtccaccac caggctctcc tagatgtgaa gaacatcgcc caccagaaca
     4441 agtttgggga ggatatgccc cacccacacg gatccgacta caaagaccat gacggtgatt
     4501 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     4561 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4621 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4681 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4741 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4801 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4861 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4921 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     4981 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5041 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5101 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     5161 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5221 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     5281 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5341 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5401 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     5461 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5521 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5581 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     5641 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5701 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5761 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5821 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     5881 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     6001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6061 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     6181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6241 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     6301 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6361 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6421 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6481 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6541 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6601 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6661 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6721 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6781 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6841 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6901 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6961 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7021 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     7081 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     7141 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     7201 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     7261 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7321 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7381 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     7441 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7501 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7561 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7621 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7681 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7741 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7801 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7861 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7921 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7981 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8041 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8101 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8161 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8221 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8281 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8341 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8401 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8461 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8521 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8581 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8641 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8701 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8761 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8821 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8881 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8941 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9001 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     9061 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9121 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9181 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9241 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9301 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9361 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9421 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9481 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9541 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9601 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     9661 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     9721 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     9781 ctgcttagg
//
