LOCUS       V047250                 8074 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047250
VERSION     V047250
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..3894
                     /label="Dis3(NM_028315)"
                     /note="Dis3(NM_028315)"
                     /gene="Dis3"
     polyA_signal    3938..4162
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4208..4636
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4650..4979
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4991..5590
                     /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    5750..5871
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5920..5936)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5944..5960
                     /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(5968..5998)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6013..6034
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6322..6907)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7078..7938)
                     /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(7939..8043)
                     /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 atgctcaggt ccaagacgtt cttgaagaag acccgcgcgg gcggcgtggt gaagatcgtg
     1081 cgcgagcact acctgcggga tgacatcggc tgcggcgcgc cggcttgctc ggcctgcggc
     1141 ggggcgcacg cgggcccggc cctggagctg cagccccggg accaggcgag cagcctctgc
     1201 ccgtggccgc actaccttct gccggacacc aatgtgctgc tgcaccagat tgatgtcctc
     1261 gaacacccgg ccatcagaaa tgtcattgtg ctacaaacag tgatgcaaga agtgagaaac
     1321 cggagcgccc ccatctacaa gcgaatcagg gatgtgacca ataaccagga aaagcatttc
     1381 tataccttca ctaatgagca ccataaagaa acctacatcg agcaagagca gggagagaat
     1441 gccaatgaca ggaatgacag agccatccga gtcgcagcga agtggtacaa cgagcacctg
     1501 aagagggtgg cagcagacag tcagctgcaa gttatcctga taaccaatga caggaagaac
     1561 aaagagaaag ctgtgcaaga ggggatacca gccttcacgt gtgaagaata cgtaaagagc
     1621 ctgactgcta accctgaact tatagaccgt cttgcttact tgtccgatga aatgaatgaa
     1681 atagaaagtg ggaaaataat attttcagag catcttccct taagcaagct ccaacaaggc
     1741 ataaaatctg gttcctatct tcaaggaaca ttcagagcta gcagggaaaa ttatttggag
     1801 gctacagtat ggattcatgg agacaaagaa gaggaaaaag agatacttat acagggaatt
     1861 aagcatctaa acagagctgt gcatgaagac attgtggccg tggagctact gcccaggagc
     1921 cagtgggtgg caccgtcttc cgtggtttta gacgatgaag gtcaaaatga agacgatgtg
     1981 gagaaagatg aggagagaga actcctgctt aagactgctg taagtgaaaa aatgttacgg
     2041 cctacaggtc gagttgtggg gataataaaa aggaactgga gaccgtattg tggcatgctt
     2101 tccaagtctg atattaagga gtcaagaaga catctcttta cacccgctga taagagaatt
     2161 ccacgaattc ggatagaaac cagacaggct tctgcgttag aaggacggag aattattgtc
     2221 gctattgatg gttggcctag aaattctaga tatccaaatg gacactttgt aaagaattta
     2281 ggcgatgttg gagagaagga gacagaaacg gaagtgttgc tgctcgagca cgatgttcct
     2341 catcagccct tttcccaggc tgtgcttagc ttcctgccca ggatgccctg gagcattact
     2401 gaggaggaca tgaaaaaccg agaagacctg agacatctgt gtgtttgcag tgtggaccct
     2461 ccagggtgca ctgacataga tgacgctctg cattgtagag agctcagcaa tggaaacttg
     2521 gaggttggtg ttcatattgc ggatgttagc cattttatca ggccaggaaa tgcgttggat
     2581 caagaatctg caagaagagg aacaactgtt tatctttgtg aaaagaggat tgacatggtt
     2641 ccagagttgc tcagctccaa cctctgttcc ttaagatcca acgttgacag gttggcattt
     2701 tcctgtattt gggaaatgaa tcataatgct gaaatattaa aaacgagatt taccaaaagt
     2761 gtcattaatt caaaggcttc tcttacgtac gcggaagcac agatgagaat tgattcggcg
     2821 gctatgaatg atgatattac cactagtctc cgtggactca atcagctggc taaaattcta
     2881 aaaaagggaa ggattgaaaa gggggctttg actctgtctt ctccagagat ccgattccac
     2941 atggacagtg aaacccacga cccaatcgac ctgcagacga aagagctgag agaaacaaat
     3001 tccatggtgg aagaatttat gttacttgct aatatttctg tcgcaaaaaa aattcatgaa
     3061 gagttttctg aacatgctct gcttcggaaa cacccagctc cgcctccctc gaattatgac
     3121 attcttgtta aggctgcaaa gtccaagaat ttgcaaatta agactgatac agccaaatct
     3181 ttggccgact ctttggaccg ggctgaatct ccggatttcc catacctgaa tactctatta
     3241 aggatactgg ccactcgctg tatgatgcaa gctgtgtact tctgctctgg gatggataat
     3301 gattttcatc actatggctt agcctccccc atatacacac atttcacctc tcctatcaga
     3361 agatatgcag acataattgt ccatcggcta ttagctgtgg cgattggggc tgactgtact
     3421 taccctgagt tgacagacaa acacaagctt tcagatatat gtaaaaacct caatttccgg
     3481 cataaaatgg ctcagtatgc ccagcgtgct tcggtggctt ttcatacaca gttgtttttc
     3541 aaaagcaaag gaatagtaag tgaagaagcc tatattctct ttgtaaggaa aaatgcaatt
     3601 gtggtgttaa ttccaaagta tggcttagaa ggtacagttt tctttgaaga aaaagataaa
     3661 ccaaagccac gccttgctta cgatgatgag atcccttccc tgagaataga aggtacagtg
     3721 ttccatgtgt ttgataaggt taaagtgaag atcacattag attcatcaaa tcttcaacat
     3781 cagaaaatcc gcatggccct tgtagaacca cagatcccag gaataaatat tcctcctaat
     3841 gttgcagaca aggctcttac tgcaccaggg ggaaagaaga ggaagcttga gaagtagctc
     3901 gagtgaccgg taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3961 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4021 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4081 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4141 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4201 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4261 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4321 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4381 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4441 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4501 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4561 gatttataag ggattttggc catttcggcc tattggttaa aaaatgagct gatttaacaa
     4621 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4681 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4741 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4801 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4861 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4921 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4981 agctcccggg atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     5041 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     5101 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     5161 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     5221 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     5281 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     5341 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     5401 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     5461 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     5521 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     5581 cggtgcctga ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5641 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5701 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5761 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5821 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5881 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5941 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6001 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6061 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6121 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6181 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6721 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6841 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7381 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7501 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7681 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8041 gcgcacattt ccccgaaaag tgccacctga cgtc
//
