LOCUS       V028276                 9188 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028276
VERSION     V028276
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             872..1579
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1616..4819
                     /label="PIK3CA(NM_006218)"
                     /note="PIK3CA(NM_006218)"
                     /gene="PIK3CA"
     CDS             4826..4852
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4856..4882
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4886..4912
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4950..5174
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5220..5648
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5662..5991
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6058..6657
                     /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    6831..6952
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7001..7017)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7025..7041
                     /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(7049..7079)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7094..7115
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7403..7988)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8159..9019)
                     /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(9020..9124)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagccac catggtgagc aagggcgagg aggataacat
      901 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      961 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1021 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1081 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     1141 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1201 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1261 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1321 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     1381 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     1441 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     1501 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1561 catggacgag ctgtacaagg gaggtggagg tagtggagga ggcggatcaa gcttcatgcc
     1621 tccacgacca tcatcaggtg aactgtgggg catccacttg atgcccccaa gaatcctagt
     1681 agaatgttta ctaccaaatg gaatgatagt gactttagaa tgcctccgtg aggctacatt
     1741 aataaccata aagcatgaac tatttaaaga agcaagaaaa taccccctcc atcaacttct
     1801 tcaagatgaa tcttcttaca ttttcgtaag tgttactcaa gaagcagaaa gggaagaatt
     1861 ttttgatgaa acaagacgac tttgtgacct tcggcttttt caaccctttt taaaagtaat
     1921 tgaaccagta ggcaaccgtg aagaaaagat cctcaatcga gaaattggtt ttgctatcgg
     1981 catgccagtg tgtgaatttg atatggttaa agatccagaa gtacaggact tccgaagaaa
     2041 tattctgaac gtttgtaaag aagctgtgga tcttagggac ctcaattcac ctcatagtag
     2101 agcaatgtat gtctatcctc caaatgtaga atcttcacca gaattgccaa agcacatata
     2161 taataaatta gataaagggc aaataatagt ggtgatctgg gtaatagttt ctccaaataa
     2221 tgacaagcag aagtatactc tgaaaatcaa ccatgactgt gtaccagaac aagtaattgc
     2281 tgaagcaatc aggaaaaaaa ctcgaagtat gttgctatcc tctgaacaac taaaactctg
     2341 tgttttagaa tatcagggca agtatatttt aaaagtgtgt ggatgtgatg aatacttcct
     2401 agaaaaatat cctctgagtc agtataagta tataagaagc tgtataatgc ttgggaggat
     2461 gcccaatttg atgttgatgg ctaaagaaag cctttattct caactgccaa tggactgttt
     2521 tacaatgcca tcttattcca gacgcatttc cacagctaca ccatatatga atggagaaac
     2581 atctacaaaa tccctttggg ttataaatag tgcactcaga ataaaaattc tttgtgcaac
     2641 ctacgtgaat gtaaatattc gagacattga taagatctat gttcgaacag gtatctacca
     2701 tggaggagaa cccttatgtg acaatgtgaa cactcaaaga gtaccttgtt ccaatcccag
     2761 gtggaatgaa tggctgaatt atgatatata cattcctgat cttcctcgtg ctgctcgact
     2821 ttgcctttcc atttgctctg ttaaaggccg aaagggtgct aaagaggaac actgtccatt
     2881 ggcatgggga aatataaact tgtttgatta cacagacact ctagtatctg gaaaaatggc
     2941 tttgaatctt tggccagtac ctcatggatt agaagatttg ctgaacccta ttggtgttac
     3001 tggatcaaat ccaaataaag aaactccatg cttagagttg gagtttgact ggttcagcag
     3061 tgtggtaaag ttcccagata tgtcagtgat tgaagagcat gccaattggt ctgtatcccg
     3121 agaagcagga tttagctatt cccacgcagg actgagtaac agactagcta gagacaatga
     3181 attaagggaa aatgacaaag aacagctcaa agcaatttct acacgagatc ctctctctga
     3241 aatcactgag caggagaaag attttctatg gagtcacaga cactattgtg taactatccc
     3301 cgaaattcta cccaaattgc ttctgtctgt taaatggaat tctagagatg aagtagccca
     3361 gatgtattgc ttggtaaaag attggcctcc aatcaaacct gaacaggcta tggaacttct
     3421 ggactgtaat tacccagatc ctatggttcg aggttttgct gttcggtgct tggaaaaata
     3481 tttaacagat gacaaacttt ctcagtattt aattcagcta gtacaggtcc taaaatatga
     3541 acaatatttg gataacttgc ttgtgagatt tttactgaag aaagcattga ctaatcaaag
     3601 gattgggcac tttttctttt ggcatttaaa atctgagatg cacaataaaa cagttagcca
     3661 gaggtttggc ctgcttttgg agtcctattg tcgtgcatgt gggatgtatt tgaagcacct
     3721 gaataggcaa gtcgaggcaa tggaaaagct cattaactta actgacattc tcaaacagga
     3781 gaagaaggat gaaacacaaa aggtacagat gaagttttta gttgagcaaa tgaggcgacc
     3841 agatttcatg gatgctctac agggctttct gtctcctcta aaccctgctc atcaactagg
     3901 aaacctcagg cttgaagagt gtcgaattat gtcctctgca aaaaggccac tgtggttgaa
     3961 ttgggagaac ccagacatca tgtcagagtt actgtttcag aacaatgaga tcatctttaa
     4021 aaatggggat gatttacggc aagatatgct aacacttcaa attattcgta ttatggaaaa
     4081 tatctggcaa aatcaaggtc ttgatcttcg aatgttacct tatggttgtc tgtcaatcgg
     4141 tgactgtgtg ggacttattg aggtggtgcg aaattctcac actattatgc aaattcagtg
     4201 caaaggcggc ttgaaaggtg cactgcagtt caacagccac acactacatc agtggctcaa
     4261 agacaagaac aaaggagaaa tatatgatgc agccattgac ctgtttacac gttcatgtgc
     4321 tggatactgt gtagctacct tcattttggg aattggagat cgtcacaata gtaacatcat
     4381 ggtgaaagac gatggacaac tgtttcatat agattttgga cactttttgg atcacaagaa
     4441 gaaaaaattt ggttataaac gagaacgtgt gccatttgtt ttgacacagg atttcttaat
     4501 agtgattagt aaaggagccc aagaatgcac aaagacaaga gaatttgaga ggtttcagga
     4561 gatgtgttac aaggcttatc tagctattcg acagcatgcc aatctcttca taaatctttt
     4621 ctcaatgatg cttggctctg gaatgccaga actacaatct tttgatgaca ttgcatacat
     4681 tcgaaagacc ctagccttag ataaaactga gcaagaggct ttggagtatt tcatgaaaca
     4741 aatgaatgat gcacatcatg gtggctggac aacaaaaatg gattggatct tccacacaat
     4801 taaacagcat gcattgaacc tcgagtaccc atacgacgta cctgattatg caacttatcc
     4861 ttatgatgta cctgattatg ctagctatcc ttatgatgtc ccagactacg catagtgagg
     4921 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4981 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5041 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5101 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5161 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     5221 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5281 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5341 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5401 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5461 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5521 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5581 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5641 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5821 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5881 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5941 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     6001 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     6061 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc
     6121 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc
     6181 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     6241 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     6301 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     6361 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     6421 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6481 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6541 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     6601 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagcg
     6661 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6721 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6781 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     6841 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6901 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6961 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     7021 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7081 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7141 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7201 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7261 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7321 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7921 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7981 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8041 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8101 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8161 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8221 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8281 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8341 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8401 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8461 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8521 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8581 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8641 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8701 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8761 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8821 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8881 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8941 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9001 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9061 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9121 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     9181 cctatggt
//
