LOCUS       Exported                8410 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBabe puro HA PIK3CA E545K
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8410)
  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 8410)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8410
                     /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             1377..1403
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4649..4669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4654..4983
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4834..4969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4896..4915
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4993..5592
                     /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(4993..5012)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5489..5509
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5691..6284
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6485..6502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6656..7244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6736..6755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7422..8282)
                     /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     8045..8064
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8283..8387)
                     /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 ggccggatcc aagcttcacc atggggtacc
     1381 catacgacgt gccagattac gccagatccc agtgtggtgg tacgtaccct ccaagaccat
     1441 catcaggtga actgtggggc atccacttga tgcccccaag aatcctagta gaatgtttac
     1501 taccaaatgg aatgatagtg actttagaat gcctccgtga ggctacatta ataaccataa
     1561 agcatgaact atttaaagaa gcaagaaaat accccctcca tcaacttctt caagatgaat
     1621 cttcttacat tttcgtaagt gttactcaag aagcagaaag ggaagaattt tttgatgaaa
     1681 caagacgact ttgtgacctt cggctttttc aacccttttt aaaagtaatt gaaccagtag
     1741 gcaaccgtga agaaaagatc ctcaatcgag aaattggttt tgctatcggc atgccagtgt
     1801 gtgaatttga tatggttaaa gatccagaag tacaggactt ccgaagaaat gttctgaacg
     1861 tttgtaaaga agctgtggat cttagggacc tcaattcacc tcatagtaga gcaatgtatg
     1921 tctatcctcc aaatgtagaa tcttcaccag aattgccaaa gcacatatat aataaattag
     1981 ataaagggca aataatagtg gtgatctggg taatagtttc tccaaataat gacaagcaga
     2041 agtatactct gaaaatcaac catgactgtg taccagaaca agtaattgct gaagcaatca
     2101 ggaaaaaaac tcgaagtatg ttgctatcct ctgaacaact aaaactctgt gttttagaat
     2161 atcagggcaa gtatatttta aaagtgtgtg gatgtgatga atacttccta gaaaaatatc
     2221 ctctgagtca gtataagtat ataagaagct gtataatgct tgggaggatg cccaatttga
     2281 tgttgatggc taaagaaagc ctttattctc aactgccaat ggactgtttt acaatgccat
     2341 cttattccag acgcatttcc acagctacac catatatgaa tggagaaaca tctacaaaat
     2401 ccctttgggt tataaatagt gcactcagaa taaaaattct ttgtgcaacc tacgtgaatg
     2461 taaatattcg agacattgat aagatctatg ttcgaacagg tatctaccat ggaggagaac
     2521 ccttatgtga caatgtgaac actcaaagag taccttgttc caatcccagg tggaatgaat
     2581 ggctgaatta tgatatatac attcctgatc ttcctcgtgc tgctcgactt tgcctttcca
     2641 tttgctctgt taaaggccga aagggtgcta aagaggaaca ctgtccattg gcatggggaa
     2701 atataaactt gtttgattac acagacactc tagtatctgg aaaaatggct ttgaatcttt
     2761 ggccagtacc tcatggatta gaagatttgc tgaaccctat tggtgttact ggatcaaatc
     2821 caaataaaga aactccatgc ttagagttgg agtttgactg gttcagcagt gtggtaaagt
     2881 tcccagatat gtcagtgatt gaagagcatg ccaattggtc tgtatcccga gaagcaggat
     2941 ttagctattc ccacgcagga ctgagtaaca gactagctag agacaatgaa ttaagggaaa
     3001 atgacaaaga acagctcaaa gcaatttcta cacgagatcc tctctctgaa atcactaagc
     3061 aggagaaaga ttttctatgg agtcacagac actattgtgt aactatcccc gaaattctac
     3121 ccaaattgct tctgtctgtt aaatggaatt ctagagatga agtagcccag atgtattgct
     3181 tggtaaaaga ttggcctcca atcaaacctg aacaggctat ggaacttctg gactgtaatt
     3241 acccagatcc tatggttcga ggttttgctg ttcggtgctt ggaaaaatat ttaacagatg
     3301 acaaactttc tcagtattta attcagctag tacaggtcct aaaatatgaa caatatttgg
     3361 ataacttgct tgtgagattt ttactgaaga aagcattgac taatcaaagg attgggcact
     3421 ttttcttttg gcatttaaaa tctgagatgc acaataaaac agttagccag aggtttggcc
     3481 tgcttttgga gtcctattgt cgtgcatgtg ggatgtattt gaagcacctg aataggcaag
     3541 tcgaggcaat ggaaaagctc attaacttaa ctgacattct caaacaggag aagaaggatg
     3601 aaacacaaaa ggtacagatg aagtttttag ttgagcaaat gaggcgacca gatttcatgg
     3661 atgctctaca gggctttctg tctcctctaa accctgctca tcaactagga aacctcaggc
     3721 ttgaagagtg tcgaattatg tcctctgcaa aaaggccact gtggttgaat tgggagaacc
     3781 cagacatcat gtcagagtta ctgtttcaga acaatgagat catctttaaa aatggggatg
     3841 atttacggca agatatgcta acacttcaaa ttattcgtat tatggaaaat atctggcaaa
     3901 atcaaggtct tgatcttcga atgttacctt atggttgtct gtcaatcggt gactgtgtgg
     3961 gacttattga ggtggtgcga aattctcaca ctattatgca aattcagtgc aaaggcggct
     4021 tgaaaggtgc actgcagttc aacagccaca cactacatca gtggctcaaa gacaagaaca
     4081 aaggagaaat atatgatgca gccattgacc tgtttacacg ttcatgtgct ggatactgtg
     4141 tagctacctt cattttggga attggagatc gtcacaatag taacatcatg gtgaaagacg
     4201 atggacaact gtttcatata gattttggac actttttgga tcacaagaag aaaaaatttg
     4261 gttataaacg agaacgtgtg ccatttgttt tgacacagga tttcttaata gtgattagta
     4321 aaggagccca agaatgcaca aagacaagag aatttgagag gtttcaggag atgtgttaca
     4381 aggcttatct agctattcga cagcatgcca atctcttcat aaatcttttc tcaatgatgc
     4441 ttggctctgg aatgccagaa ctacaatctt ttgatgacat tgcatacatt cgaaagaccc
     4501 tagccttaga taaaactgag caagaggctt tggagtattt catgaaacaa atgaatgatg
     4561 cacatcatgg tggctggaca acaaaaatgg attggatctt ccacacaatt aaacagcatg
     4621 cattgaactg agtcgacctc gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4681 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4741 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4801 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4861 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4921 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4981 aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     5041 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     5101 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     5161 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     5221 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5281 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5341 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5401 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5461 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5521 accgtcaccg ccgacgtcga gtgcccgaag gaccgcgcga cctggtgcat gacccgcaag
     5581 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     5641 catgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     5701 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     5761 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat
     5821 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     5881 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     5941 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
     6001 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     6061 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     6121 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
     6181 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     6241 tctcctctga gtgattgact acccgtcagc gggggtcttt catttccgac ttgtggtctc
     6301 gctgccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tcttcacatg
     6361 cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag
     6421 ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg cgctcttccg
     6481 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6541 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6601 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6661 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6721 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6781 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6841 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6901 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6961 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7021 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7081 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7141 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7201 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7261 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7321 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt ttgcggccgc
     7381 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7441 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7501 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7561 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7621 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7681 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7741 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7801 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7861 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7921 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7981 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     8041 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8101 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8161 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8221 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8281 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8341 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8401 aaagtgccac
//
