LOCUS       V032306                 6586 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032306
VERSION     V032306
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"
     CDS             937..2337
                     /label="KAT8(NM_182958)"
                     /note="KAT8(NM_182958)"
                     /gene="KAT8"
     CDS             2350..2415
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2450..2674
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2720..3148
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3162..3491
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3503..4102
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4262..4383
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4432..4448)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4456..4472
                     /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(4480..4510)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4525..4546
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4834..5419)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5590..6450)
                     /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(6451..6555)
                     /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 aagctttcta gagaattcga tatcggatcc gccaccatgg cggcacaggg agctgctgcg
      961 gcggttgcgg cggggacttc aggggtcgcg ggggagggcg agcccgggcc cggggagaat
     1021 gcggccgctg aggggaccgc cccatccccg ggccgcgtct ctccgccgac cccggcgcgc
     1081 ggcgagccgg aagtcacggt ggagatcgga gaaacgtacc tgtgccggcg accggatagc
     1141 acctggcatt ctgctgaagt gatccagtct cgagtgaacg accaggaggg ccgagaggaa
     1201 ttctatgtac actacgtggg ctttaaccgg cggctggacg agtgggtaga caagaaccgg
     1261 ctggcgctga ccaagacagt gaaggatgct gtacagaaga actcagagaa gtacctgagc
     1321 gagctcgcag agcagcctga gcgcaagatc actcgcaacc aaaagcgcaa gcatgatgag
     1381 atcaaccatg tgcagaagac ttatgcagag atggacccca ccacagcagc cttggagaag
     1441 gagcatgagg cgatcaccaa ggtgaagtat gtggacaaga tccacatcgg gaactacgaa
     1501 attgatgcct ggtatttctc accattcccc gaagactatg ggaaacagcc caagctctgg
     1561 ctctgcgagt actgcctcaa gtacatgaaa tatgagaaga gctaccgctt ccacttgggt
     1621 cagtgccagt ggcggcagcc ccccgggaaa gagatctacc gcaagagcaa catctccgtg
     1681 tacgaagttg atggcaaaga ccataagatt tactgtcaga acctgtgtct gctggccaag
     1741 cttttcctgg accataagac actgtacttt gacgtggagc cgttcgtctt ttacatcctg
     1801 actgaggtgg accggcaggg ggcccacatt gttggctact tctccaagga gaaggagtcc
     1861 ccggatggaa acaatgtggc ctgcatcctg accttgcccc cctaccaacg ccgcggctac
     1921 gggaagttcc tcatcgcttt cagttatgag ctctccaagc tggagagcac agtcggctcc
     1981 ccggagaagc cactgtctga cctgggcaag ctcagctacc gcagctactg gtcctgggtg
     2041 ctgctagaga tcctgcggga cttccggggc acactgtcca tcaaggacct cagccagatg
     2101 accagtatca cccaaaatga catcatcagt accctgcaat ccctcaatat ggtcaagtac
     2161 tggaagggcc agcacgtgat ctgtgtcaca cccaagctgg tggaggagca cctcaaaagt
     2221 gcccagtata agaaaccacc catcacaggt gggtgggggg ctgctgtgtg tcgggggcgg
     2281 tgggggagtg tcagtatatg gactggtagg agtcaaggcc tccttattgc tgtcacactc
     2341 gagaccggtg actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag
     2401 gatgacgatg acaagtgacc ggtaagttta aaccgctgat cagcctcgac tgtgccttct
     2461 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2521 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2581 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2641 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2701 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2761 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2821 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2881 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2941 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3001 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3061 gtctattctt ttgatttata agggattttg gccatttcgg cctattggtt aaaaaatgag
     3121 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3241 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3301 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3361 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3421 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3481 gcttttgcaa aaagctcccg ggatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3541 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3601 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3661 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3721 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3781 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3841 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3901 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     3961 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4021 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4081 gacccgcaag cccggtgcct gattcgaaat gaccgaccaa gcgacgccca acctgccatc
     4141 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4201 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4261 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4321 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4381 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4441 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4501 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4561 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4621 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4681 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4741 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4801 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4861 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4921 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4981 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5041 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5101 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5161 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5221 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5281 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5341 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5401 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5461 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5521 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5581 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5641 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5701 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5761 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5821 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5881 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5941 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6001 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6061 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6121 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6181 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6241 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6301 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6361 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6421 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6481 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6541 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
