LOCUS       V044694                 8245 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044694
VERSION     V044694
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             928..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             988..1014
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1021..4065
                     /label="Hira(NM_010435)"
                     /note="Hira(NM_010435)"
                     /gene="Hira"
     polyA_signal    4109..4333
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4379..4807
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4821..5150
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5162..5761
                     /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    5921..6042
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6091..6107)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6115..6131
                     /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(6139..6169)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6184..6205
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6493..7078)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7249..8109)
                     /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(8110..8214)
                     /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 gagaattcgc caccatgtac ccatacgacg tacctgatta tgcaacttat
      961 ccttatgatg tacctgatta tgctagttat ccttatgatg tcccagacta cgcaggatcc
     1021 atgaagctct tgaagccaac ctgggtcaac cacaatggga agccaatttt ttcagttgat
     1081 attcaccctg atgggaccaa gtttgcaact ggaggacaag ggcaggattc tgggaaggtt
     1141 gtgatctgga atatgtctcc agtcctccag gaggatgacg agaaggatga aaatattccc
     1201 aagatgcttt gccagatgga caatcactta gcatgtgtga actgtgtgcg gtggtcaaac
     1261 agtgggatgt atttagcttc tgggggagat gacaaactga ttatggtgtg gaagcgggct
     1321 acgtacattg ggcccagcac tgtgtttggt tccagtggta agcttgccaa tgtggagcaa
     1381 tggcggtgtg tctccatcct ccggagtcac tcaggcgatg tgatggatgt agcatggtct
     1441 ccccacgacg cctggctggc ctcatgcagc gtggataaca ctgttgtcat ttggaatgcc
     1501 gtgaagttcc cagaaattct tgcaactctg agaggtcatt ctggcttagt aaaaggtttg
     1561 acttgggatc ccgttggtaa atatattgcc tctcaagctg atgatcgaag tttgaaggta
     1621 tggaggacgc tggactggca gctagagact agcatcacca agccttttga tgagtgtgga
     1681 ggaacgacgc atgttctccg gcttagttgg tcacctgatg gccattacct ggtatctgcc
     1741 catgccatga ataattctgg ccccactgct cagatcatcg aaagagaggg ctggaagacc
     1801 aacatggact ttgtgggtca ccggaaagct gtgactgttg tgaaattcaa cccaaaaatc
     1861 ttcaagaaga agcagaagaa tgggagctct acaaagccca gctgcccata ctgctgctgt
     1921 gctgttggca gcaaggaccg ctcactctct gtctggctca catgtttgaa acggcctctg
     1981 gttgtcatcc atgaactgtt tgacaagtcc atcatggata tttcctggac tctgaatggg
     2041 ttgggtatcc tggtgtgctc catggacggc tctgtggcgt tccttgattt ctctcaggat
     2101 gaactcggag accccctgag tgaggaggaa aagagccgaa ttcaccagtc tacctatggc
     2161 aagagcctgg caataatgac tgaggcccag ctttccacag ctgttattga gaaccctgag
     2221 atgctcaagt accagcggag acagcaacag cagcagctgg atcagaagaa tgccactact
     2281 agggagacaa gctcagcatc ctcagtcacg ggtgtggtca atggggaaag tctagaagat
     2341 atcagaaaga atcttttgaa gaaacaagtg gaaactcgga cagcagatgg tcggaggaga
     2401 atcacgcctc tttgcatagc acagctggac actggggact tctccacggc attcttcaac
     2461 agcatcccac tctccagctc cctagcaggc accatgctct cctctcctag tggtcagcag
     2521 ctactaccac tggactccag taccccctcc ttcggcgcct caaagccttg cacagaacca
     2581 gtggcagcca ccagtgccag gcctacaggc gaatctgtca gtaaggacag tatgaatgct
     2641 acctctactc ctgctgcatc gtcaccctct gtgttaacaa ccccatccaa gattgaaccc
     2701 atgaaagcat ttgattcccg gttcacagaa cggtccaaag ccacaccagg tgctccttcc
     2761 ttgaccagtg tgattccaac agctgttgaa aggttgaaag agcagaacct cgtcaaggag
     2821 ctgaggtccc gggaactgga gagcagcagt gacagcgatg agaaggtcca cctagccaag
     2881 ccctcttcac tgtccaagcg caaacttgag cttgaggtag agacggtgga aaagaagaag
     2941 aaaggtcgcc ctaggaagga ttcacgtctt ttacccatgt ctctgtccgt ccagtctcca
     3001 gctgccctgt ctacagagaa ggaggccatg tgtctgtctg caccagcact tgcactgaag
     3061 ctgccaattc caggcccaca gagagcgttc accctccagg tgagctctga cccctccatg
     3121 tacattgagg tggagaatga agtgaccacc gttgggggga taaggctgag tcgcctgaag
     3181 tgcaaccgtg aagggaagga gtgggagaca gtgctcagca gtcgcgtcct cactgctgcc
     3241 ggcagctgtg atgtggtatg tgttgcctgt gaaaagagga tgctgtcggt gttctctacc
     3301 tgtggtcgcc gtctcctccc tcccatcctc cttccatctc caatctctac tttgcactgc
     3361 acgggcccct acgtcatggc actcaccgct gcagccacac tgtctgtctg ggatgttcac
     3421 agacaggtgg ttgtggtgaa agaagaatct ctacactcca tcttgtcagg aagtgatatg
     3481 acggtgtcac agatcttgct aacacagcat ggaatcccag tgatgaattt gtctgatggg
     3541 aaagcatact gctttaatcc atccctctcc acatggaacc tggtttctga caagcaagat
     3601 tcattggccc agtgtgcaga cttcaggaac agcctgccgt cccaggatgc catgctttgt
     3661 tcaggaccgt tagccataat tcagggccgc acctccaact ctggaaggca agctgcccgg
     3721 ctcttctccg tgcctcatgt ggtacagcag gagaccacac tggcctacct agagaaccag
     3781 gttgctgcgg cacttaccct gcagtcaagc cacgagtatc gtcactggct cctcctttat
     3841 gctcggtacc ttgtgaatga agggtttgaa taccgcctcc gtgaaatatg caaggacttg
     3901 ctgggtccag ttcactgctc cactggaagt cagtgggagt caacagtagt gggtctgcgg
     3961 aaaagggagc tgctgaagga actgttgcca gtcattgggc agaatcttcg attccagcgc
     4021 ctctttactg agtgccagga acaactggac atcctgagag acaagtagct cgagtgaccg
     4081 gtaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4141 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4201 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4261 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4321 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4381 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4441 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4501 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4561 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4621 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4681 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4741 gggattttgg ccatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4801 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4861 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4921 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4981 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5041 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5101 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5161 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     5221 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     5281 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     5341 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     5401 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     5461 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     5521 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     5581 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     5641 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     5701 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     5761 attcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5821 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5881 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6001 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6061 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6121 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6181 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6241 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6301 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6361 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6421 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6481 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6541 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6601 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6661 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6721 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6781 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6841 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6901 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6961 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7021 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7081 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7141 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7201 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7261 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7321 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7381 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7441 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7501 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7561 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7621 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7681 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7741 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7801 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7861 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7921 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7981 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8041 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8101 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8161 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8221 tccccgaaaa gtgccacctg acgtc
//
