LOCUS Exported 8725 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Retroviral construct for expressing human EGFR with L858R mutation in human cells. ACCESSION . VERSION . KEYWORDS EGFR L858R 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8725 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1..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" primer_bind 3917..3936 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" 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(6352..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" primer_bind 6975..6994 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" 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 //