LOCUS       Exported                8261 bp ds-DNA     circular SYN 12-JUL-2022
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 8261)
  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 8261)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 12, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8261
                     /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        4707..4917
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4729..4925
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4776..4911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4838..4857
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4935..5534
                     /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(4935..4954)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5431..5451
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5755..6224
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6354..6371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6525..7113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6605..6624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7273..8133)
                     /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     7896..7915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8134..8238)
                     /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 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4801 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4861 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4921 gcaaaaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     4981 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5041 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5101 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5161 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     5221 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5281 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5341 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5401 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5461 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5521 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
     5581 cccatgcatc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     5641 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     5701 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat
     5761 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     5821 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     5881 cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc
     5941 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     6001 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     6061 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     6121 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     6181 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca
     6241 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     6301 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6361 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6421 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6481 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6541 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6601 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6661 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6721 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6781 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6841 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6901 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6961 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7021 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7081 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7141 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7201 aggatcttca cctagatcct tttgcggccg caaatcaatc taaagtatat atgagtaaac
     7261 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7321 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7381 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7441 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7501 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7561 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7621 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7681 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7741 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7801 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7861 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7921 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7981 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8041 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8101 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8161 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8221 acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
