LOCUS       Exported                8725 bp ds-DNA     circular SYN 23-SEP-2022
DEFINITION  Retroviral construct for expressing human EGFR in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    EGFR WT
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 Sep 23, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8725
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             23..492
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    557..756
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             957..1373
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1303..1325
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1341..1357
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1401..5033
                     /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
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     primer_bind     complement(5070..5090)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5075..5404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5255..5390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5317..5336
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5414..6013
                     /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(5414..5433)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5910..5930
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6234..6703
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6833..6850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7004..7592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7084..7103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7752..8612)
                     /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     8375..8394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8613..8717)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca gcctgaatat gggccaaaca ggatatctgt ggtaagcagt
       61 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
      121 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
      181 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
      241 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
      301 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct
      361 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
      421 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
      481 gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga
      541 cccaccaccg ggaggcaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
      601 ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc
      661 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
      721 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
      781 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
      841 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
      901 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
      961 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1021 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1081 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1141 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1201 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1261 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1321 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggccgga
     1381 tcccagtgtg gtggtacatc atgcgaccct ccgggacggc cggggcagcg ctcctggcgc
     1441 tgctggctgc gctctgcccg gcgagtcggg ctctggagga aaagaaagtt tgccaaggca
     1501 cgagtaacaa gctcacgcag ttgggcactt ttgaagatca ttttctcagc ctccagagga
     1561 tgttcaataa ctgtgaggtg gtccttggga atttggaaat tacctatgtg cagaggaatt
     1621 atgatctttc cttcttaaag accatccagg aggtggctgg ttatgtcctc attgccctca
     1681 acacagtgga gcgaattcct ttggaaaacc tgcagatcat cagaggaaat atgtactacg
     1741 aaaattccta tgccttagca gtcttatcta actatgatgc aaataaaacc ggactgaagg
     1801 agctgcccat gagaaattta caggaaatcc tgcatggcgc cgtgcggttc agcaacaacc
     1861 ctgccctgtg caacgtggag agcatccagt ggcgggacat agtcagcagt gactttctca
     1921 gcaacatgtc gatggacttc cagaaccacc tgggcagctg ccaaaagtgt gatccaagct
     1981 gtcccaatgg gagctgctgg ggtgcaggag aggagaactg ccagaaactg accaaaatca
     2041 tctgtgccca gcagtgctcc gggcgctgcc gtggcaagtc ccccagtgac tgctgccaca
     2101 accagtgtgc tgcaggctgc acaggccccc gggagagcga ctgcctggtc tgccgcaaat
     2161 tccgagacga agccacgtgc aaggacacct gccccccact catgctctac aaccccacca
     2221 cgtaccagat ggatgtgaac cccgagggca aatacagctt tggtgccacc tgcgtgaaga
     2281 agtgtccccg taattatgtg gtgacagatc acggctcgtg cgtccgagcc tgtggggccg
     2341 acagctatga gatggaggaa gacggcgtcc gcaagtgtaa gaagtgcgaa gggccttgcc
     2401 gcaaagtgtg taacggaata ggtattggtg aatttaaaga ctcactctcc ataaatgcta
     2461 cgaatattaa acacttcaaa aactgcacct ccatcagtgg cgatctccac atcctgccgg
     2521 tggcatttag gggtgactcc ttcacacata ctcctcctct ggatccacag gaactggata
     2581 ttctgaaaac cgtaaaggaa atcacagggt ttttgctgat tcaggcttgg cctgaaaaca
     2641 ggacggacct ccatgccttt gagaacctag aaatcatacg cggcaggacc aagcaacatg
     2701 gtcagttttc tcttgcagtc gtcagcctga acataacatc cttgggatta cgctccctca
     2761 aggagataag tgatggagat gtgataattt caggaaacaa aaatttgtgc tatgcaaata
     2821 caataaactg gaaaaaactg tttgggacct ccggtcagaa aaccaaaatt ataagcaaca
     2881 gaggtgaaaa cagctgcaag gccacaggcc aggtctgcca tgccttgtgc tcccccgagg
     2941 gctgctgggg cccggagccc agggactgcg tctcttgccg gaatgtcagc cgaggcaggg
     3001 aatgcgtgga caagtgcaac cttctggagg gtgagccaag ggagtttgtg gagaactctg
     3061 agtgcataca gtgccaccca gagtgcctgc ctcaggccat gaacatcacc tgcacaggac
     3121 ggggaccaga caactgtatc cagtgtgccc actacattga cggcccccac tgcgtcaaga
     3181 cctgcccggc aggagtcatg ggagaaaaca acaccctggt ctggaagtac gcagacgccg
     3241 gccatgtgtg ccacctgtgc catccaaact gcacctacgg atgcactggg ccaggtcttg
     3301 aaggctgtcc aacgaatggg cctaagatcc cgtccatcgc cactgggatg gtgggggccc
     3361 tcctcttgct gctggtggtg gccctgggga tcggcctctt catgcgaagg cgccacatcg
     3421 ttcggaagcg cacgctgcgg aggctgctgc aggagaggga gcttgtggag cctcttacac
     3481 ccagtggaga agctcccaac caagctctct tgaggatctt gaaggaaact gaattcaaaa
     3541 agatcaaagt gctgggctcc ggtgcgttcg gcacggtgta taagggactc tggatcccag
     3601 aaggtgagaa agttaaaatt cccgtcgcta tcaaggaatt aagagaagca acatctccga
     3661 aagccaacaa ggaaatcctc gatgaagcct acgtgatggc cagcgtggac aacccccacg
     3721 tgtgccgcct gctgggcatc tgcctcacct ccaccgtgca actcatcacg cagctcatgc
     3781 ccttcggctg cctcctggac tatgtccggg aacacaaaga caatattggc tcccagtacc
     3841 tgctcaactg gtgtgtgcag atcgcaaagg gcatgaacta cttggaggac cgtcgcttgg
     3901 tgcaccgcga cctggcagcc aggaacgtac tggtgaaaac accgcagcat gtcaagatca
     3961 cagattttgg gctggccaaa ctgctgggtg cggaagagaa agaataccat gcagaaggag
     4021 gcaaagtgcc tatcaagtgg atggcattgg aatcaatttt acacagaatc tatacccacc
     4081 agagtgatgt ctggagctac ggggtgaccg tttgggagtt gatgaccttt ggatccaagc
     4141 catatgacgg aatccctgcc agcgagatct cctccatcct ggagaaagga gaacgcctcc
     4201 ctcagccacc catatgtacc atcgatgtct acatgatcat ggtcaagtgc tggatgatag
     4261 acgcagatag tcgcccaaag ttccgtgagt tgatcatcga attctccaaa atggcccgag
     4321 acccccagcg ctaccttgtc attcaggggg atgaaagaat gcatttgcca agtcctacag
     4381 actccaactt ctaccgtgcc ctgatggatg aagaagacat ggacgacgtg gtggatgccg
     4441 acgagtacct catcccacag cagggcttct tcagcagccc ctccacgtca cggactcccc
     4501 tcctgagctc tctgagtgca accagcaaca attccaccgt ggcttgcatt gatagaaatg
     4561 ggctgcaaag ctgtcccatc aaggaagaca gcttcttgca gcgatacagc tcagacccca
     4621 caggcgcctt gactgaggac agcatagacg acaccttcct cccagtgcct gaatacataa
     4681 accagtccgt tcccaaaagg cccgctggct ctgtgcagaa tcctgtctat cacaatcagc
     4741 ctctgaaccc cgcgcccagc agagacccac actaccagga cccccacagc actgcagtgg
     4801 gcaaccccga gtatctcaac actgtccagc ccacctgtgt caacagcaca ttcgacagcc
     4861 ctgcccactg ggcccagaaa ggcagccacc aaattagcct ggacaaccct gactaccagc
     4921 aggacttctt tcccaaggaa gccaagccaa atggcatctt taagggctcc acagctgaaa
     4981 atgcagaata cctaagggtc gcgccacaaa gcagtgaatt tattggagca tgagatgtag
     5041 gaattcgcca gcacagtggt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5101 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5161 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5221 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5281 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5341 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5401 caaaaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     5461 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     5521 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     5581 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5641 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     5701 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     5761 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     5821 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     5881 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     5941 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     6001 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
     6061 ccatgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     6121 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     6181 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata
     6241 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     6301 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     6361 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     6421 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     6481 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     6541 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     6601 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     6661 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa
     6721 aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa
     6781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6841 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6901 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7441 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7561 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7621 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7681 ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact
     7741 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7801 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7861 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7921 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7981 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8041 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8101 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8161 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8221 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8281 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8341 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8401 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8461 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8521 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8581 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8641 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8701 caaatagggg ttccgcgcac atttc
//
