LOCUS       V039752                 9381 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039752
VERSION     V039752
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4061
                     /label="Pik3cg(NM_001371300)"
                     /note="Pik3cg(NM_001371300)"
                     /gene="Pik3cg"
     CDS             4068..4133
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4157..4745
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4811..4859
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5061..5489
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5503..5832
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5899..6693
                     /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    6869..6990
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7039..7055)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7063..7079
                     /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(7087..7117)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7132..7153
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7441..8029)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8200..9060)
                     /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(9061..9165)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga gctggagaac tatgaacaac
      781 ctgtggttct gagagaggac aacctccgca ggcgccggag gatgaagcca cgcagcgcag
      841 caggcagcct gtcttccatg gagctcatcc ccattgagtt cgtactgccc accagccagc
      901 gcatcagcaa gacaccggaa acggccctgc tgcatgtggc tggccacggc aatgtggagc
      961 agatgaaagc tcaggtgtgg ttacgcgcac tggagaccag tgtggctgcg gaattctacc
     1021 accgattagg tccggaccaa ttcctcctcc tctaccagaa gaagggacag tggtatgaga
     1081 tctacgatag gtaccaagtg gtgcagaccc tagactgcct acgttactgg aagatgatgc
     1141 acaagagccc cggccagatc cacgtggtac agcgacatgt accttctgag gagaccctgg
     1201 ctttccagaa gcagctcacc tccctgattg gctacgacgt cactgacatc agcaacgtgc
     1261 acgatgatga gctagaattc actcgccgcc gtctggttac gccccgcatg gctgaagtgg
     1321 ctggccggga tgccaaactc tatgccatgc acccttgggt gacctccaaa cctcttccag
     1381 actacctgtc aaaaaaaatt gtcaacaatt gcattttcat tgtcatccat cgtggtacca
     1441 ccagtcaaac catcaaggtc tccgcagatg atactcctga caccatcctt cagagcttct
     1501 tcaccaagat ggccaagaag aagtccctaa tgaatatccc agaaagtcaa agtgagcagg
     1561 attttgtgct gcgggtctgt ggccgtgatg agtacctagt aggtgaaacg cccctcaaaa
     1621 atttccagtg ggtgaggcag tgcctcaaga acggagagga aatacacctg gtgctcgaca
     1681 cgcctccaga cccggcccta gatgaggtga ggaacgaaga atggccgctg gtggatgact
     1741 gcactggagt caccggctac catgagcagc tgaccatcca tggcaaggac cacgagagcg
     1801 ttttcacagt gtctttgtgg gactgtgacc gaaagttcag ggtcaagatc agaggcattg
     1861 atatccctgt cttgcctcgg aacaccgacc tcactgtttt tgtggaagca aatatccagc
     1921 atgggcaaca agtcctctgc caaaggagaa ccagccctaa gctcttcaca gaagaggtgc
     1981 tctggaatgt gtggctggag tttggcatta aaatcaaaga cttgcccaaa ggggccctac
     2041 tgaacctaca gatctactgc tgcaaaaccc catcactgtc cagtaaggct tccgtagaga
     2101 cgccaggctc cgagtccaag ggcaaagccc agctcctcta ttacgtgaac ttgctgttga
     2161 tagaccaccg cttcctcctc cgccatgggg gctacgtgct ccacatgtgg cagatatctg
     2221 ggaaggcaga ggaacagggc agcttcaatg ccgacaagct cacctccgca accaatcctg
     2281 acaaggagaa ctcaatgtcc atttccatct tgctggacaa ctactgtcac cccattgctt
     2341 tgcctaagca ccggcctact ccggacccag aaggagacag agttcgagct gaaatgccca
     2401 atcagcttcg aaagcaattg gaggcaatca tagccacaga tccacttaac cccctcacag
     2461 cagaggacaa agaattgctc tggcatttta gatatgaaag cctgaagcat ccaaaggcat
     2521 accctaaact attcagctca gtaaagtggg gacagcaaga aattgttgcc aaaacatacc
     2581 agctattagc cagaagggag atctgggatc aaagtgcttt ggatgttggc ttaaccatgc
     2641 aactcctgga ctgcaacttt tcagatgaaa acgtaagggc cattgcagtt cagaaactgg
     2701 agagcttgga ggacgatgac gttttacatt accttctcca gctggtgcag gctgtgaaat
     2761 ttgaaccata ccatgacagt gcgctggcca gattcttgct gaagcgtggc ttgagaaaca
     2821 agagaattgg tcacttctta ttctggttct tgcgaagtga gatagcccag tccagacact
     2881 atcagcagag gttcgctgtg atcctggaag cgtacctgcg cggctgtggc aaggccatgc
     2941 tacaggactt cacgcagcag gtccacgtga ttgagatgtt acagaaagtc accattgata
     3001 tcaaatcact ctcggctgag aagtatgatg tcagttccca agttatttca cagcttaagc
     3061 aaaagcttga aagcctgcag aattccagtc ttcctgagag ctttagagtt ccctatgatc
     3121 ctggactgaa agcaggtacc ctggtgattg agaagtgtaa agtgatggcc tccaagaaga
     3181 agcccctgtg gcttgagttt aaacgtgccg atcctacagt cctatccaat gaaaccattg
     3241 ggatcatctt taaacatggc gacgatctac gccaagacat gttgatctta cagattctac
     3301 gcatcatgga gtccatttgg gaggctgaat ctctggacct gtgccttctg ccttacggtt
     3361 gcatctcaac tggtgacaaa atagggatga tagagattgt gaaggacgcc acgacgattg
     3421 ctcaaattca gcagagcacg gtgggtaaca cgggggcatt caaagatgaa gtcctgaatc
     3481 actggctcaa ggaaaaatgc cctattgaag aaaagtttca ggcagcagtg gaaaggttcg
     3541 tttactcctg tgcaggctac tgtgtggcca catttgttct tgggataggt gacaggcaca
     3601 acgacaacat catgatctca gagacaggaa acctatttca catagacttt ggacacattc
     3661 ttggcaatta caagagtttc ctgggcatca ataaagagag agtacctttt gtcctaaccc
     3721 cggacttctt gtttgtgatg ggaacttcgg gcaaaaagac aagtccacac ttccagaaat
     3781 tccaggatgt ctgcgttagg gcttacctag ctcttcgcca ccacacaaac ctgctgatca
     3841 tcctgttctc catgatgctg atgacaggaa tgccgcagct gacaagcaaa gaggacattg
     3901 aatacatccg ggatgccctc atggtgggga gaagcgagga ggacgccaag aagtatttcc
     3961 ttgatcagat tgaagtctgc agggacaaag gatggactgt gcagtttaac tggttcctcc
     4021 atcttgttct tggcatcaag caaggagaga aacactccgc tggatccgac tacaaagacc
     4081 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     4141 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4201 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4261 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4321 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4381 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4441 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4501 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     4561 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4621 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4681 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4741 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4801 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     4861 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4921 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4981 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     5041 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5101 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5161 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     5221 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5281 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5341 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5401 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     5461 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5521 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5581 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5641 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5701 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5761 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5821 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5881 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5941 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6001 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6061 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6121 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6181 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6241 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6301 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6361 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6421 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6481 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6541 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6601 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6661 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     6721 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6781 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6841 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6901 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6961 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     7021 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     7081 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     7141 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     7201 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7261 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7321 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7621 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7861 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     8161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     8221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8281 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8341 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8401 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8461 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8521 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8581 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8641 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8701 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8761 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8821 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8881 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8941 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     9001 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     9061 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     9121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     9181 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     9241 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     9301 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     9361 tgcatgaaga atctgcttag g
//
