LOCUS       Exported                8451 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBabe puro Myr HA PIK3CA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  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 8451)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8451
                     /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             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1364..1405
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1424..1450
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4690..4710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4695..5024
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4875..5010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4937..4956
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5034..5633
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(5034..5053)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5530..5550
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5732..6325
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6526..6543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6697..7285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6777..6796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7463..8323)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8086..8105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8324..8428)
                     /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 taagctggcc 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 tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccggatct accatggggt cttcaaaatc
     1381 taaaccaaag gaccccagcc agcgccggcg caggatccga ggttacccat acgacgtgcc
     1441 agattacgcc agatcccagt gtggtggtac gtaccctcca agaccatcat caggtgaact
     1501 gtggggcatc cacttgatgc ccccaagaat cctagtagaa tgtttactac caaatggaat
     1561 gatagtgact ttagaatgcc tccgtgaggc tacattaata accataaagc atgaactatt
     1621 taaagaagca agaaaatacc ccctccatca acttcttcaa gatgaatctt cttacatttt
     1681 cgtaagtgtt actcaagaag cagaaaggga agaatttttt gatgaaacaa gacgactttg
     1741 tgaccttcgg ctttttcaac cctttttaaa agtaattgaa ccagtaggca accgtgaaga
     1801 aaagatcctc aatcgagaaa ttggttttgc tatcggcatg ccagtgtgtg aatttgatat
     1861 ggttaaagat ccagaagtac aggacttccg aagaaatgtt ctgaacgttt gtaaagaagc
     1921 tgtggatctt agggacctca attcacctca tagtagagca atgtatgtct atcctccaaa
     1981 tgtagaatct tcaccagaat tgccaaagca catatataat aaattagata aagggcaaat
     2041 aatagtggtg atctgggtaa tagtttctcc aaataatgac aagcagaagt atactctgaa
     2101 aatcaaccat gactgtgtac cagaacaagt aattgctgaa gcaatcagga aaaaaactcg
     2161 aagtatgttg ctatcctctg aacaactaaa actctgtgtt ttagaatatc agggcaagta
     2221 tattttaaaa gtgtgtggat gtgatgaata cttcctagaa aaatatcctc tgagtcagta
     2281 taagtatata agaagctgta taatgcttgg gaggatgccc aatttgatgt tgatggctaa
     2341 agaaagcctt tattctcaac tgccaatgga ctgttttaca atgccatctt attccagacg
     2401 catttccaca gctacaccat atatgaatgg agaaacatct acaaaatccc tttgggttat
     2461 aaatagtgca ctcagaataa aaattctttg tgcaacctac gtgaatgtaa atattcgaga
     2521 cattgataag atctatgttc gaacaggtat ctaccatgga ggagaaccct tatgtgacaa
     2581 tgtgaacact caaagagtac cttgttccaa tcccaggtgg aatgaatggc tgaattatga
     2641 tatatacatt cctgatcttc ctcgtgctgc tcgactttgc ctttccattt gctctgttaa
     2701 aggccgaaag ggtgctaaag aggaacactg tccattggca tggggaaata taaacttgtt
     2761 tgattacaca gacactctag tatctggaaa aatggctttg aatctttggc cagtacctca
     2821 tggattagaa gatttgctga accctattgg tgttactgga tcaaatccaa ataaagaaac
     2881 tccatgctta gagttggagt ttgactggtt cagcagtgtg gtaaagttcc cagatatgtc
     2941 agtgattgaa gagcatgcca attggtctgt atcccgagaa gcaggattta gctattccca
     3001 cgcaggactg agtaacagac tagctagaga caatgaatta agggaaaatg acaaagaaca
     3061 gctcaaagca atttctacac gagatcctct ctctgaaatc actgagcagg agaaagattt
     3121 tctatggagt cacagacact attgtgtaac tatccccgaa attctaccca aattgcttct
     3181 gtctgttaaa tggaattcta gagatgaagt agcccagatg tattgcttgg taaaagattg
     3241 gcctccaatc aaacctgaac aggctatgga acttctggac tgtaattacc cagatcctat
     3301 ggttcgaggt tttgctgttc ggtgcttgga aaaatattta acagatgaca aactttctca
     3361 gtatttaatt cagctagtac aggtcctaaa atatgaacaa tatttggata acttgcttgt
     3421 gagattttta ctgaagaaag cattgactaa tcaaaggatt gggcactttt tcttttggca
     3481 tttaaaatct gagatgcaca ataaaacagt tagccagagg tttggcctgc ttttggagtc
     3541 ctattgtcgt gcatgtggga tgtatttgaa gcacctgaat aggcaagtcg aggcaatgga
     3601 aaagctcatt aacttaactg acattctcaa acaggagaag aaggatgaaa cacaaaaggt
     3661 acagatgaag tttttagttg agcaaatgag gcgaccagat ttcatggatg ctctacaggg
     3721 ctttctgtct cctctaaacc ctgctcatca actaggaaac ctcaggcttg aagagtgtcg
     3781 aattatgtcc tctgcaaaaa ggccactgtg gttgaattgg gagaacccag acatcatgtc
     3841 agagttactg tttcagaaca atgagatcat ctttaaaaat ggggatgatt tacggcaaga
     3901 tatgctaaca cttcaaatta ttcgtattat ggaaaatatc tggcaaaatc aaggtcttga
     3961 tcttcgaatg ttaccttatg gttgtctgtc aatcggtgac tgtgtgggac ttattgaggt
     4021 ggtgcgaaat tctcacacta ttatgcaaat tcagtgcaaa ggcggcttga aaggtgcact
     4081 gcagttcaac agccacacac tacatcagtg gctcaaagac aagaacaaag gagaaatata
     4141 tgatgcagcc attgacctgt ttacacgttc atgtgctgga tactgtgtag ctaccttcat
     4201 tttgggaatt ggagatcgtc acaatagtaa catcatggtg aaagacgatg gacaactgtt
     4261 tcatatagat tttggacact ttttggatca caagaagaaa aaatttggtt ataaacgaga
     4321 acgtgtgcca tttgttttga cacaggattt cttaatagtg attagtaaag gagcccaaga
     4381 atgcacaaag acaagagaat ttgagaggtt tcaggagatg tgttacaagg cttatctagc
     4441 tattcgacag catgccaatc tcttcataaa tcttttctca atgatgcttg gctctggaat
     4501 gccagaacta caatcttttg atgacattgc atacattcga aagaccctag ccttagataa
     4561 aactgagcaa gaggctttgg agtatttcat gaaacaaatg aatgatgcac atcatggtgg
     4621 ctggacaaca aaaatggatt ggatcttcca cacaattaaa cagcatgcat tgaactgagt
     4681 cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4741 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4861 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4921 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4981 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt accatgaccg
     5041 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     5101 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     5161 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5221 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5281 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     5341 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     5401 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5461 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5521 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5581 agtgcccgaa ggaccgcgcg acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     5641 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg ataaaataaa
     5701 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     5761 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     5821 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
     5881 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     5941 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     6001 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     6061 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     6121 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     6181 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     6241 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     6301 tacccgtcag cgggggtctt tcatttccga cttgtggtct cgctgccttg ggagggtctc
     6361 ctctgagtga ttgactaccc gtcagcgggg gtcttcacat gcagcatgta tcaaaattaa
     6421 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     6481 aacgcgcggg gagaggcggt ttgcgtattg gcgctcttcc gcttcctcgc tcactgactc
     6541 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6601 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6661 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6721 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6781 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6841 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6901 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6961 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7021 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7081 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7141 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7201 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7261 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7321 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7381 cacctagatc cttttaaatt aaaaatgaag tttgcggccg caaatcaatc taaagtatat
     7441 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7501 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7561 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7621 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7681 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7741 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7801 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7861 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7921 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7981 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8041 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     8101 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8161 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8221 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8281 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8341 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8401 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
