LOCUS       V036338                10396 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036338
VERSION     V036338
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             931..5832
                     /label="PIK3C2B(NM_001377334)"
                     /note="PIK3C2B(NM_001377334)"
                     /gene="PIK3C2B"
     CDS             5839..5928
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    5963..6187
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6233..6661
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6675..7004
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7071..7865
                     /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    8039..8160
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8209..8225)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8233..8249
                     /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(8257..8287)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8302..8323
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8611..9196)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9367..10227)
                     /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(10228..10332)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgtcttcga ctcagggcaa tggggaacac
      961 tggaagtccc tggagtcagt gggcatcagc cgcaaagagc tagcgatggc cgaagccctg
     1021 cagatggagt atgatgccct gtcccggctc cggcatgaca aggaggagaa cagagccaag
     1081 cagaacgcag acccctctct catcagctgg gatgagcctg gggtagactt ttacagcaag
     1141 ccagcaggaa ggcggaccga cctcaagctg ttacgcggtc tctctggctc tgatcctacc
     1201 cttaactaca actcactctc cccacaggaa gggccgccca accactctac ctcccaaggg
     1261 ccacagcctg gctcagatcc ctggcccaaa ggctccctgt ctggagacta tctctacatt
     1321 tttgatggtt cagatggggg agtctcttcg tccccaggac caggggacat agagggctct
     1381 tgcaagaaac tatccccacc tcctctgcct ccccgagctt ctatctggga tacccctccc
     1441 ctgcctccca gaaaggggtc cccctcatcc tccaagatct cccagcccag tgacatcaac
     1501 actttctctt tggtcgaaca attgccgggc aaactgctag agcatcggat cctagaagag
     1561 gaagaggtgc tgggaggtgg gggtcagggg cgcctactgg ggtctgtgga ctatgatggt
     1621 atcaatgatg caattactag gctcaacttg aaatcgacct atgatgcgga gatgttgcgg
     1681 gatgccacca ggggctggaa ggagggccga gggccgctgg acttcagcaa agacacctct
     1741 ggaaaacccg tggccaggag caagactatg ccccctcagg tgcccccccg cacctatgcc
     1801 tcccgctatg gcaaccgaaa gaatgcgacg cctggcaaga accgccggat ttctgcagcc
     1861 ccggtgggct cccggcccca cactgttgcc aatggccatg agttgtttga ggtctcagaa
     1921 gagagagatg aggaggttgc tgcattttgc cacatgctgg atatccttcg atctggctct
     1981 gacatccaag actacttcct cactggctat gtctggagtg ctgtcacccc tagcccagag
     2041 cacctcgggg atgaggtcaa cctgaaggtg actgtgttgt gtgacaggct tcaagaggca
     2101 ctcactttca cctgcaactg ttcctccact gtagacttgc ttatctacca gaccctgtgc
     2161 tacacccatg atgacctgag gaatgtggac gtgggtgact ttgtgctaaa gccctgcggg
     2221 ctggaggagt tcctgcagaa caagcatgcc ttgggcagtc atgagtacat ccaatactgc
     2281 cgcaagtttg acattgacat tcggctacag ctgatggagc agaaggttgt gcgcagtgac
     2341 ctggcccgga cggtgaatga tgaccagagc ccctccacct tgaactacct cgtccatctc
     2401 caagagaggc ctgtcaagca gaccatcagc aggcaggccc tgagtcttct gttcgacact
     2461 taccacaatg aggtggatgc cttcctgctg gctgatggag acttcccact gaaggctgac
     2521 agggtggtcc agtccgtcaa ggccatctgc aacgccctgg ccgccgtgga aacccctgag
     2581 atcaccagtg ctctcaacca gctgcccccc tgcccctccc gcatgcagcc taaaattcag
     2641 aaggatccca gtgtcttggc tgtgagggaa aaccgagaga aggtcgtgga agccctgacc
     2701 gctgccatct tggacctggt ggagctgtac tgcaacacat tcaacgcaga cttccagacg
     2761 gcagtgcccg ggagccgcaa gcatgacctg gtccaggagg cctgccattt cgccaggtcc
     2821 ctggccttca ctgtctatgc cacccaccgc atccccatca tctgggctac cagctatgaa
     2881 gatttctacc tctcctgctc cctcagccat ggcggcaagg agctgtgcag ccccctgcag
     2941 acccgaagag ctcacttctc caagtacctc ttccacctca tcgtctggga ccagcagatc
     3001 tgcttcccag tgcaggtgaa ccggctgcct cgggagacac tgctgtgtgc cactctctat
     3061 gctctgccca tccccccacc ggggagctcc tcagaggcca ataagcagcg gcgggtgcct
     3121 gaagccctgg gctgggtcac taccccactc ttcaacttca ggcaggtcct gacctgtggc
     3181 cggaagcttc tgggtttgtg gccagcaaca caggaaaatc ccagcgcccg ttggagtgca
     3241 cctaatttcc accagccaga cagtgtcatc ctgcagattg acttccccac ctcggccttt
     3301 gacatcaagt tcaccagccc ccctggagac aagttcagcc cccgctatga gtttggcagc
     3361 ctccgggaag aagaccagcg caagcttaaa gacatcatgc agaaagagtc cttgtactgg
     3421 ctcactgatg ctgacaagaa gcgcctgtgg gagaagcgat attactgcca ctcggaggtg
     3481 agctcgctcc ccctggtgct cgccagcgcc cccagctggg agtgggcttg cctgcctgac
     3541 atctatgttc tcctgaagca gtggacccac atgaaccacc aggatgccct ggggctcctg
     3601 catgccacct tcccggacca ggaggtgcgt cgtatggctg tgcagtggat tggctcactc
     3661 tcagatgctg agctgctaga ctacctgccc cagctggtac aggccctgaa gtatgaatgc
     3721 tacctggaca gcccgttggt gcgcttcctc ctgaaacgag ctgtgtctga cttgagagtg
     3781 actcactact tcttctggtt actgaaggac ggcctcaagg actctcagtt cagcatccgc
     3841 taccagtatc tgctggcagc cttactgtgc tgctgtggca aggggctgag agaagagttt
     3901 aaccgccagt gctggcttgt caatgccctg gccaaactgg cccagcaggt ccgggaggca
     3961 gccccatctg caaggcaggg aatcctccgc acgggcctgg aggaggtgaa gcagttcttt
     4021 gccctcaatg gctcgtgccg cttgccactc agccccagtc tgctggttaa gggaattgtg
     4081 cccagggact gttcctactt caactccaat gctgtccccc tcaaactctc cttccaaaat
     4141 gtggatcccc tgggtgagaa catccgtgtc atcttcaagt gtggggacga ccttcgccag
     4201 gacatgctaa cgctgcagat gattcgcatc atgagcaaga tctgggtcca ggaggggctg
     4261 gacatgcgca tggtcatctt ccgctgcttc tccaccggcc ggggcagagg gatggtggag
     4321 atgatcccta atgctgagac cctgcgtaag atccaggtgg agcatggggt gaccggctcg
     4381 ttcaaggacc ggcccctggc agactggctg cagaaacaca accctgggga ggacgagtat
     4441 gagaaggctg tggagaactt tatctactcc tgcgctggct gctgcgtggc cacgtacgtc
     4501 ttgggcatct gtgaccgaca taatgacaac atcatgctga agaccactgg tcacatgttc
     4561 cacattgatt ttggccgctt cctgggccat gcccagatgt ttggcaacat caagcgggac
     4621 cgtgccccct ttgtcttcac ctcggacatg gcgtatgtca tcaacggggg tgacaagcct
     4681 tccagccgct tccatgattt tgttgacctt tgctgccaag cctacaacct cattcgcaag
     4741 cacacccacc tcttcctcaa ccttctgggc ctgatgttgt cctgtgggat ccctgaactc
     4801 tcagacctgg aggacctcaa gtatgtgtac gatgccctga ggcctcagga tacagaggcc
     4861 aatgccacta cctacttcac taggttgatt gagtccagcc tgggcagtgt agccacaaag
     4921 ctcaattttt tcatccataa tctggctcag atgaagttca cgggctcaga tgaccggctg
     4981 accctctcct ttgcctcccg aacacacact ctcaagagct ctggccgaat cagtgatgtt
     5041 ttcctctgcc gccatgagaa gatcttccac cccaacaaag gctatatata tgtggtaaag
     5101 gtgatgcgag agaacactca cgaggccacc tacatccagc ggacctttga ggagttccag
     5161 gaattacaca ataagttgcg gctgctcttc ccttcttccc acttgcccag cttccctagt
     5221 cgcttcgtga tcggccgctc ccggggagag gcggtggccg agcggcggag ggaggagcta
     5281 aacggttaca tctggcactt gatccacgca ccccctgagg tggccgagtg tgatttggtg
     5341 tacaccttct tccacccact gccccgggat gagaaggcta tgggcaccag cccagctcct
     5401 aagtcctcag atggcacatg ggcccggccc gtcggaaagg tgggagggga ggtgaagctg
     5461 tccatctcct acaaaaacaa taaactcttc atcatggtga tgcatattcg gggcttgcaa
     5521 ctgctccagg atggaaatga ccctgacccc tatgtgaaaa tttacctcct tcctgaccct
     5581 cagaaaacca ctaagaggaa aaccaaagtg gcccggaaaa cctgcaatcc tacctacaat
     5641 gagatgttgg tatatgatgg gatccccaag ggtgacctgc agcagcggga gctccagctg
     5701 agcgtgctga gtgagcaggg attctgggag aacgtcctcc tcggtgaggt gaacatccgc
     5761 ctgcgagagc tggacctggc tcaggagaag accggctggt tcgccctggg atctcgaagt
     5821 catggcacct tgctcgagga acaaaaacta atatcagagg aagatttgga acagaaacta
     5881 atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttatg agggcccgtt
     5941 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6001 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     6061 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     6121 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     6181 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     6241 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6301 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6361 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6421 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6481 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6541 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6601 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6661 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     6721 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     6781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     6841 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6901 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     6961 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     7021 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     7081 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     7141 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     7201 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     7261 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     7321 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     7381 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     7441 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     7501 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     7561 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     7621 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     7681 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     7741 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     7801 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     7861 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     7921 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     7981 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     8041 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8101 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8161 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     8221 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     8281 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     8341 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     8401 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     8461 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     8521 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     8581 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     8641 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     8701 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     8761 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     8821 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     8881 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     8941 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     9001 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     9061 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     9121 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     9181 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     9241 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     9301 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     9361 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     9421 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     9481 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     9541 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     9601 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     9661 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     9721 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     9781 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     9841 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     9901 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     9961 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    10021 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    10081 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    10141 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    10201 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    10261 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
    10321 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
    10381 cccgatcccc tatggt
//
