LOCUS       sequence-163488-        8725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral construct for expressing human EGFR with L858R mutation 
            in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8725)
  AUTHORS   Greulich H, Chen TH, Feng W, Janne PA, Alvarez JV, Zappaterra M, 
            Bulmer SE, Frank DA, Hahn WC, Sellers WR, Meyerson M
  TITLE     Oncogenic Transformation by Inhibitor-Sensitive and -Resistant EGFR 
            Mutants.
  JOURNAL   PLoS Med. 2005 Oct 4. 2(11):e313.
  PUBMED    16187797
REFERENCE   2  (bases 1 to 8725)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Med. 
            2005 Oct 4. 2(11):e313."
COMMENT     Sequence Label: EGFR L858R
FEATURES             Location/Qualifiers
     source          1..8725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..72,73..1935,1936..2004,2005..3633)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGRAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     primer_bind     complement(3670..3690)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3675..4004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3855..3990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4014..4613
                     /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(4014..4033)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4510..4530
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4834..5303
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5433..5450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5604..6192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5684..5703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6352..7143,7144..7212))
                     /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"
     promoter        complement(7213..7317)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             7348..7817
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7882..8081
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             8282..8698
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8628..8650
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8666..8682
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 atgcgaccct ccgggacggc cggggcagcg ctcctggcgc tgctggctgc gctctgcccg
       61 gcgagtcggg ctctggagga aaagaaagtt tgccaaggca cgagtaacaa gctcacgcag
      121 ttgggcactt ttgaagatca ttttctcagc ctccagagga tgttcaataa ctgtgaggtg
      181 gtccttggga atttggaaat tacctatgtg cagaggaatt atgatctttc cttcttaaag
      241 accatccagg aggtggctgg ttatgtcctc attgccctca acacagtgga gcgaattcct
      301 ttggaaaacc tgcagatcat cagaggaaat atgtactacg aaaattccta tgccttagca
      361 gtcttatcta actatgatgc aaataaaacc ggactgaagg agctgcccat gagaaattta
      421 caggaaatcc tgcatggcgc cgtgcggttc agcaacaacc ctgccctgtg caacgtggag
      481 agcatccagt ggcgggacat agtcagcagt gactttctca gcaacatgtc gatggacttc
      541 cagaaccacc tgggcagctg ccaaaagtgt gatccaagct gtcccaatgg gagctgctgg
      601 ggtgcaggag aggagaactg ccagaaactg accaaaatca tctgtgccca gcagtgctcc
      661 gggcgctgcc gtggcaagtc ccccagtgac tgctgccaca accagtgtgc tgcaggctgc
      721 acaggccccc gggagagcga ctgcctggtc tgccgcaaat tccgagacga agccacgtgc
      781 aaggacacct gccccccact catgctctac aaccccacca cgtaccagat ggatgtgaac
      841 cccgagggca aatacagctt tggtgccacc tgcgtgaaga agtgtccccg taattatgtg
      901 gtgacagatc acggctcgtg cgtccgagcc tgtggggccg acagctatga gatggaggaa
      961 gacggcgtcc gcaagtgtaa gaagtgcgaa gggccttgcc gcaaagtgtg taacggaata
     1021 ggtattggtg aatttaaaga ctcactctcc ataaatgcta cgaatattaa acacttcaaa
     1081 aactgcacct ccatcagtgg cgatctccac atcctgccgg tggcatttag gggtgactcc
     1141 ttcacacata ctcctcctct ggatccacag gaactggata ttctgaaaac cgtaaaggaa
     1201 atcacagggt ttttgctgat tcaggcttgg cctgaaaaca ggacggacct ccatgccttt
     1261 gagaacctag aaatcatacg cggcaggacc aagcaacatg gtcagttttc tcttgcagtc
     1321 gtcagcctga acataacatc cttgggatta cgctccctca aggagataag tgatggagat
     1381 gtgataattt caggaaacaa aaatttgtgc tatgcaaata caataaactg gaaaaaactg
     1441 tttgggacct ccggtcagaa aaccaaaatt ataagcaaca gaggtgaaaa cagctgcaag
     1501 gccacaggcc aggtctgcca tgccttgtgc tcccccgagg gctgctgggg cccggagccc
     1561 agggactgcg tctcttgccg gaatgtcagc cgaggcaggg aatgcgtgga caagtgcaac
     1621 cttctggagg gtgagccaag ggagtttgtg gagaactctg agtgcataca gtgccaccca
     1681 gagtgcctgc ctcaggccat gaacatcacc tgcacaggac ggggaccaga caactgtatc
     1741 cagtgtgccc actacattga cggcccccac tgcgtcaaga cctgcccggc aggagtcatg
     1801 ggagaaaaca acaccctggt ctggaagtac gcagacgccg gccatgtgtg ccacctgtgc
     1861 catccaaact gcacctacgg atgcactggg ccaggtcttg aaggctgtcc aacgaatggg
     1921 cctaagatcc cgtccatcgc cactgggatg gtgggggccc tcctcttgct gctggtggtg
     1981 gccctgggga tcggcctctt catgcgaagg cgccacatcg ttcggaagcg cacgctgcgg
     2041 aggctgctgc aggagaggga gcttgtggag cctcttacac ccagtggaga agctcccaac
     2101 caagctctct tgaggatctt gaaggaaact gaattcaaaa agatcaaagt gctgggctcc
     2161 ggtgcgttcg gcacggtgta taagggactc tggatcccag aaggtgagaa agttaaaatt
     2221 cccgtcgcta tcaaggaatt aagagaagca acatctccga aagccaacaa ggaaatcctc
     2281 gatgaagcct acgtgatggc cagcgtggac aacccccacg tgtgccgcct gctgggcatc
     2341 tgcctcacct ccaccgtgca actcatcacg cagctcatgc ccttcggctg cctcctggac
     2401 tatgtccggg aacacaaaga caatattggc tcccagtacc tgctcaactg gtgtgtgcag
     2461 atcgcaaagg gcatgaacta cttggaggac cgtcgcttgg tgcaccgcga cctggcagcc
     2521 aggaacgtac tggtgaaaac accgcagcat gtcaagatca cagattttgg gagggccaaa
     2581 ctgctgggtg cggaagagaa agaataccat gcagaaggag gcaaagtgcc tatcaagtgg
     2641 atggcattgg aatcaatttt acacagaatc tatacccacc agagtgatgt ctggagctac
     2701 ggggtgaccg tttgggagtt gatgaccttt ggatccaagc catatgacgg aatccctgcc
     2761 agcgagatct cctccatcct ggagaaagga gaacgcctcc ctcagccacc catatgtacc
     2821 atcgatgtct acatgatcat ggtcaagtgc tggatgatag acgcagatag tcgcccaaag
     2881 ttccgtgagt tgatcatcga attctccaaa atggcccgag acccccagcg ctaccttgtc
     2941 attcaggggg atgaaagaat gcatttgcca agtcctacag actccaactt ctaccgtgcc
     3001 ctgatggatg aagaagacat ggacgacgtg gtggatgccg acgagtacct catcccacag
     3061 cagggcttct tcagcagccc ctccacgtca cggactcccc tcctgagctc tctgagtgca
     3121 accagcaaca attccaccgt ggcttgcatt gatagaaatg ggctgcaaag ctgtcccatc
     3181 aaggaagaca gcttcttgca gcgatacagc tcagacccca caggcgcctt gactgaggac
     3241 agcatagacg acaccttcct cccagtgcct gaatacataa accagtccgt tcccaaaagg
     3301 cccgctggct ctgtgcagaa tcctgtctat cacaatcagc ctctgaaccc cgcgcccagc
     3361 agagacccac actaccagga cccccacagc actgcagtgg gcaaccccga gtatctcaac
     3421 actgtccagc ccacctgtgt caacagcaca ttcgacagcc ctgcccactg ggcccagaaa
     3481 ggcagccacc aaattagcct ggacaaccct gactaccagc aggacttctt tcccaaggaa
     3541 gccaagccaa atggcatctt taagggctcc acagctgaaa atgcagaata cctaagggtc
     3601 gcgccacaaa gcagtgaatt tattggagca tgagatgtag gaattcgcca gcacagtggt
     3661 cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3721 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3841 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3901 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3961 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt accatgaccg
     4021 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     4081 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     4141 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4201 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4261 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4321 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4381 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4441 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     4501 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4561 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     4621 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg ataaaataaa
     4681 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     4741 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     4801 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
     4861 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     4921 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     4981 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     5041 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     5101 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     5161 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     5221 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     5281 tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc
     5341 ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga
     5401 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5461 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5521 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5581 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5641 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5701 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5761 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5821 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5881 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5941 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6001 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6061 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6121 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6181 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6241 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6301 ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6361 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6421 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6481 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6541 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6601 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6661 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6721 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6781 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6841 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6901 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6961 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7021 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7081 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7141 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7201 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7261 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7321 atttccccga aaagtgccac ctgcagcctg aatatgggcc aaacaggata tctgtggtaa
     7381 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     7441 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     7501 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     7561 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     7621 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     7681 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     7741 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     7801 tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc
     7861 accgacccac caccgggagg caagctggcc agcaacttat ctgtgtctgt ccgattgtct
     7921 agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat
     7981 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg
     8041 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa
     8101 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     8161 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg
     8221 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     8281 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     8341 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     8401 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     8461 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     8521 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     8581 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     8641 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg
     8701 ccggatccca gtgtggtggt acatc
//
