LOCUS       Exported                8429 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Retroviral expression of HA tagged phosphoinositide 3-kinase H1047R 
            (Tumor-derived allele).
ACCESSION   .
VERSION     .
KEYWORDS    pBabe puro HA PIK3CA H1047R
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8429)
  AUTHORS   Zhao JJ, Liu Z, Wang L, Shin E, Loda MF, Roberts TM
  TITLE     The oncogenic properties of mutant p110alpha and p110beta 
            phosphatidylinositol 3-kinases in human mammary epithelial cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Dec 20. 102(51):18443-8.
  PUBMED    16339315
REFERENCE   2  (bases 1 to 8429)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8429
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1385..1411
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4663..4683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4779..4989
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4801..4997
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4848..4983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4910..4929
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5007..5606
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5007..5026)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5503..5523
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6503..6520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6674..7262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6754..6773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7441..8301)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8064..8083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8302..8406)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccaa gcttcaccat
     1381 ggggtaccca tacgacgtgc cagattacgc cagatcccag tgtggtggta cgtaccctcc
     1441 aagaccatca tcaggtgaac tgtggggcat ccacttgatg cccccaagaa tcctagtaga
     1501 atgtttacta ccaaatggaa tgatagtgac tttagaatgc ctccgtgagg ctacattaat
     1561 aaccataaag catgaactat ttaaagaagc aagaaaatac cccctccatc aacttcttca
     1621 agatgaatct tcttacattt tcgtaagtgt tactcaagaa gcagaaaggg aagaattttt
     1681 tgatgaaaca agacgacttt gtgaccttcg gctttttcaa ccctttttaa aagtaattga
     1741 accagtaggc aaccgtgaag aaaagatcct caatcgagaa attggttttg ctatcggcat
     1801 gccagtgtgt gaatttgata tggttaaaga tccagaagta caggacttcc gaagaaatgt
     1861 tctgaacgtt tgtaaagaag ctgtggatct tagggacctc aattcacctc atagtagagc
     1921 aatgtatgtc tatcctccaa atgtagaatc ttcaccagaa ttgccaaagc acatatataa
     1981 taaattagat aaagggcaaa taatagtggt gatctgggta atagtttctc caaataatga
     2041 caagcagaag tatactctga aaatcaacca tgactgtgta ccagaacaag taattgctga
     2101 agcaatcagg aaaaaaactc gaagtatgtt gctatcctct gaacaactaa aactctgtgt
     2161 tttagaatat cagggcaagt atattttaaa agtgtgtgga tgtgatgaat acttcctaga
     2221 aaaatatcct ctgagtcagt ataagtatat aagaagctgt ataatgcttg ggaggatgcc
     2281 caatttgatg ttgatggcta aagaaagcct ttattctcaa ctgccaatgg actgttttac
     2341 aatgccatct tattccagac gcatttccac agctacacca tatatgaatg gagaaacatc
     2401 tacaaaatcc ctttgggtta taaatagtgc actcagaata aaaattcttt gtgcaaccta
     2461 cgtgaatgta aatattcgag acattgataa gatctatgtt cgaacaggta tctaccatgg
     2521 aggagaaccc ttatgtgaca atgtgaacac tcaaagagta ccttgttcca atcccaggtg
     2581 gaatgaatgg ctgaattatg atatatacat tcctgatctt cctcgtgctg ctcgactttg
     2641 cctttccatt tgctctgtta aaggccgaaa gggtgctaaa gaggaacact gtccattggc
     2701 atggggaaat ataaacttgt ttgattacac agacactcta gtatctggaa aaatggcttt
     2761 gaatctttgg ccagtacctc atggattaga agatttgctg aaccctattg gtgttactgg
     2821 atcaaatcca aataaagaaa ctccatgctt agagttggag tttgactggt tcagcagtgt
     2881 ggtaaagttc ccagatatgt cagtgattga agagcatgcc aattggtctg tatcccgaga
     2941 agcaggattt agctattccc acgcaggact gagtaacaga ctagctagag acaatgaatt
     3001 aagggaaaat gacaaagaac agctcaaagc aatttctaca cgagatcctc tctctgaaat
     3061 cactgagcag gagaaagatt ttctatggag tcacagacac tattgtgtaa ctatccccga
     3121 aattctaccc aaattgcttc tgtctgttaa atggaattct agagatgaag tagcccagat
     3181 gtattgcttg gtaaaagatt ggcctccaat caaacctgaa caggctatgg aacttctgga
     3241 ctgtaattac ccagatccta tggttcgagg ttttgctgtt cggtgcttgg aaaaatattt
     3301 aacagatgac aaactttctc agtatttaat tcagctagta caggtcctaa aatatgaaca
     3361 atatttggat aacttgcttg tgagattttt actgaagaaa gcattgacta atcaaaggat
     3421 tgggcacttt ttcttttggc atttaaaatc tgagatgcac aataaaacag ttagccagag
     3481 gtttggcctg cttttggagt cctattgtcg tgcatgtggg atgtatttga agcacctgaa
     3541 taggcaagtc gaggcaatgg aaaagctcat taacttaact gacattctca aacaggagaa
     3601 gaaggatgaa acacaaaagg tacagatgaa gtttttagtt gagcaaatga ggcgaccaga
     3661 tttcatggat gctctacagg gctttctgtc tcctctaaac cctgctcatc aactaggaaa
     3721 cctcaggctt gaagagtgtc gaattatgtc ctctgcaaaa aggccactgt ggttgaattg
     3781 ggagaaccca gacatcatgt cagagttact gtttcagaac aatgagatca tctttaaaaa
     3841 tggggatgat ttacggcaag atatgctaac acttcaaatt attcgtatta tggaaaatat
     3901 ctggcaaaat caaggtcttg atcttcgaat gttaccttat ggttgtctgt caatcggtga
     3961 ctgtgtggga cttattgagg tggtgcgaaa ttctcacact attatgcaaa ttcagtgcaa
     4021 aggcggcttg aaaggtgcac tgcagttcaa cagccacaca ctacatcagt ggctcaaaga
     4081 caagaacaaa ggagaaatat atgatgcagc cattgacctg tttacacgtt catgtgctgg
     4141 atactgtgta gctaccttca ttttgggaat tggagatcgt cacaatagta acatcatggt
     4201 gaaagacgat ggacaactgt ttcatataga ttttggacac tttttggatc acaagaagaa
     4261 aaaatttggt tataaacgag aacgtgtgcc atttgttttg acacaggatt tcttaatagt
     4321 gattagtaaa ggagcccaag aatgcacaaa gacaagagaa tttgagaggt ttcaggagat
     4381 gtgttacaag gcttatctag ctattcgaca gcatgccaat ctcttcataa atcttttctc
     4441 aatgatgctt ggctctggaa tgccagaact acaatctttt gatgacattg catacattcg
     4501 aaagacccta gccttagata aaactgagca agaggctttg gagtatttca tgaaacaaat
     4561 gaatgatgca cgtcatggtg gctggacaac aaaaatggat tggatcttcc acacaattaa
     4621 acagcatgca ttgaactgag tcgacctcga cctcgaccct gtggaatgtg tgtcagttag
     4681 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4741 agtcagcaac cataggcgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4801 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4861 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4921 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4981 gcctaggctt ttgcaaaaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca
     5041 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     5101 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     5161 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     5221 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     5281 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     5341 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5401 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5461 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5521 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5581 gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa
     5641 ggagcgcacg accccatgca tcgataaaat aaaagatttt atttagtctc cagaaaaagg
     5701 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     5761 aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg
     5821 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     5881 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     5941 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
     6001 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     6061 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     6121 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
     6181 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     6241 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggg cagttgcatc
     6301 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     6361 cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt
     6421 acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6481 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7081 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc
     7381 aaatcaatct aaagtatata tgagtaaact aagtatatat gagtaaactt ggtctgacag
     7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8401 tccgcgcaca tttccccgaa aagtgccac
//
