LOCUS       Exported                8327 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Retroviral expression of HA tagged phosphoinositide 3-kinase.
ACCESSION   .
VERSION     .
KEYWORDS    pBabe puro HA PIK3CA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8327)
  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 8327)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8327
                     /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(4657..4677)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4662..4991
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4842..4977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4904..4923
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5001..5600
                     /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(5001..5020)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5497..5517
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5821..6290
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6420..6437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6591..7179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6671..6690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7339..8199)
                     /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     7962..7981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8200..8304)
                     /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 catcatggtg gctggacaac aaaaatggat tggatcttcc acacaattaa
     4621 acagcatgca ttgaactgag tcgacctcga ccctgtggaa tgtgtgtcag ttagggtgtg
     4681 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4741 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4801 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4861 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4921 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4981 gcttttgcaa aaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     5041 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     5101 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     5161 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     5221 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     5281 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     5341 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5401 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     5461 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5521 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5581 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
     5641 cacgacccca tgcatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     5701 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     5761 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     5821 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     5881 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5941 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     6001 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     6061 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     6121 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     6181 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     6241 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     6301 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     6361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7021 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7081 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7141 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7201 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7261 tcaaaaagga tcttcaccta gatccttttg cggccgcaaa tcaatctaaa gtatatatga
     7321 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7381 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7441 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7501 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7561 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7621 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7681 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7741 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7801 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7861 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7921 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7981 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8041 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8101 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8161 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8221 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8281 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
