LOCUS       sequence-471864-        8406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EGFR(L858R,T790M)-NanoLuc(R) Fusion Protein in Mammalian 
            Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8406)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8406)
  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: 
            "Unpublished"
COMMENT     Sequence Label: EGFR(L858R,T790M)-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    828..876
                     /note="synthetic polyadenylation signal"
     CDS             complement(940..1734)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1051..1070)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1661..1680
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1766..1813)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1828..2185)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1842..1977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1994..2185
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2142..2162
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2430..2551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2467..2486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2521..2540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2588..2606
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2694..3206)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             complement(join(3261..4886,4887..4955,4956..6818,
                     6819..6890))
                     /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
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLIMQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGRAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     primer_bind     complement(6903..6922)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6904..6922)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6964..6983)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          6966..7098
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(7234..7437)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7263..7283)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7438..7817
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    8008..8291
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgggta
      721 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
      781 gacaagggaa aacgcaagcg caaagaggat ccttatcgct atcgattcac acaaaaaacc
      841 aacacacaga tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat ggcaggttgg
      901 gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag
      961 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     1021 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     1081 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     1141 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     1201 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     1261 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     1321 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     1381 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     1441 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     1501 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
     1561 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     1621 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     1681 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgatctg
     1741 cctcctgggt ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg
     1801 attaattgtc aacagttaat taagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     1861 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     1921 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     1981 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     2041 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     2101 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     2161 ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat
     2221 ttcaggccat ggtgctgcgc agatccgcgt atgcggtgtg aaataccgca cagatgcgta
     2281 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgatttta
     2341 ccacatttgt agaggtttta cttgcttaaa aaaacctccc acatctcccc ctgaacctga
     2401 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2461 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2521 gtggtttgtc caaactcatc aatgtatctt atcatgtctg ctcgaagcgg ccgccccaag
     2581 gggttatgct agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggcttt
     2641 gttagcagcc ggatcagctt gcatgcctgc aggtcgactc tagagtcgcg gccttacgcc
     2701 agaatgcgtt cgcacagccg ccagccggtc actccgttga tggttactcg gaacagcagg
     2761 gagccgtcgg ggttgatcag gcgctcgtcg ataattttgt tgccgttcca cagggtccct
     2821 gttacagtga tctttttgcc gtcgaacacg gcgatgcctt catacggccg tccgaaatag
     2881 tcgatcatgt tcggcgtaac cccgtcgatt accagtgtgc catagtgcag gatcacctta
     2941 aagtgatgat catccacagg gtacaccacc ttaaaaattt tttcgatctg gcccatttgg
     3001 tcgccgctca gaccttcata cgggatgatg acatggatgt cgatcttcag cccattttca
     3061 ccgctcagga caatcctttg gatcggagtt acggacaccc cgagattctg aaacaaactg
     3121 gacacacctc cctgttcaag gacttggtcc aggttgtagc cggctgtctg tcgccagtcc
     3181 ccaacgaaat cttcgagtgt gaagactcca ccgctgctcc ctccaccgct cccgccacca
     3241 cctgaaccac cgagagaaac tgctccaata aattcactgc tttgtggcgc gacccttagg
     3301 tattctgcat tttcagctgt ggagccctta aagatgccat ttggcttggc ttccttggga
     3361 aagaagtcct gctggtagtc agggttgtcc aggctaattt ggtggctgcc tttctgggcc
     3421 cagtgggcag ggctgtcgaa tgtgctgttg acacaggtgg gctggacagt gttgagatac
     3481 tcggggttgc ccactgcagt gctgtggggg tcctggtagt gtgggtctct gctgggcgcg
     3541 gggttcagag gctgattgtg atagacagga ttctgcacag agccagcggg ccttttggga
     3601 acggactggt ttatgtattc aggcactggg aggaaggtgt cgtctatgct gtcctcagtc
     3661 aaggcgcctg tggggtctga gctgtatcgc tgcaagaagc tgtcttcctt gatgggacag
     3721 ctttgcagcc catttctatc aatgcaagcc acggtggaat tgttgctggt tgcactcaga
     3781 gagctcagga ggggagtccg tgacgtggag gggctgctga agaagccctg ctgtgggatg
     3841 aggtactcgt cggcatccac cacgtcgtcc atgtcttctt catccatcag ggcacggtag
     3901 aagttggagt ctgtaggact tggcaaatgc attctttcat ccccctgaat gacaaggtag
     3961 cgctgggggt ctcgggccat tttggagaat tcgatgatca actcacggaa ctttgggcga
     4021 ctatctgcgt ctatcatcca gcacttgacc atgatcatgt agacatcgat ggtacatatg
     4081 ggtggctgag ggaggcgttc tcctttctcc aggatggagg agatctcgct ggcagggatt
     4141 ccgtcatatg gcttggatcc aaaggtcatc aactcccaaa cggtcacccc gtagctccag
     4201 acatcactct ggtgggtata gattctgtgt aaaattgatt ccaatgccat ccacttgata
     4261 ggcactttgc ctccttctgc atggtattct ttctcttccg cacccagcag tttggcccgc
     4321 ccaaaatctg tgatcttgac atgctgcggt gttttcacca gtacgttcct ggctgccagg
     4381 tcgcggtgca ccaagcgacg gtcctccaag tagttcatgc cctttgcgat ctgcacacac
     4441 cagttgagca ggtactggga gccaatattg tctttgtgtt cccggacata gtccaggagg
     4501 cagccgaagg gcatgagctg catgatgagt tgcacggtgg aggtgaggca gatgcccagc
     4561 aggcggcaca cgtgggggtt gtccacgctg gccatcacgt aggcttcatc gaggatttcc
     4621 ttgttggctt tcggagatgt tgcttctctt aattccttga tagcgacggg aattttaact
     4681 ttctcacctt ctgggatcca gagtccctta tacaccgtgc cgaacgcacc ggagcccagc
     4741 actttgatct ttttgaattc agtttccttc aagatcctca agagagcttg gttgggagct
     4801 tctccactgg gtgtaagagg ctccacaagc tccctctcct gcagcagcct ccgcagcgtg
     4861 cgcttccgaa cgatgtggcg ccttcgcatg aagaggccga tccccagggc caccaccagc
     4921 agcaagagga gggcccccac catcccagtg gcgatggacg ggatcttagg cccattcgtt
     4981 ggacagcctt caagacctgg ccctgtgcat ccgtaggtgc agtttggatg gcacaggtgg
     5041 cacacatggc cggcgtctgc gtacttccag accagggtgt tgttttctcc catgactcct
     5101 gccgggcagg tcttgacgca gtgggggccg tcaatgtagt gggcacactg gatacagttg
     5161 tctggtcccc gtcctgtgca ggtgatgttc atggcctgag gcaggcactc tgggtggcac
     5221 tgtatgcact cagagttctc cacaaactcc cttggctcac cctccagaag gttgcacttg
     5281 tccacgcatt ccctgcctcg gctgacattc cggcaagaga cgcagtccct gggctccggg
     5341 ccccagcagc cctcggggga gcacaaggca tggcagacct ggcctgtggc cttgcagctg
     5401 ttttcacctc tgttgcttat aattttggtt ttctgaccgg aggtcccaaa cagttttttc
     5461 cagtttattg tatttgcata gcacaaattt ttgtttcctg aaattatcac atctccatca
     5521 cttatctcct tgagggagcg taatcccaag gatgttatgt tcaggctgac gactgcaaga
     5581 gaaaactgac catgttgctt ggtcctgccg cgtatgattt ctaggttctc aaaggcatgg
     5641 aggtccgtcc tgttttcagg ccaagcctga atcagcaaaa accctgtgat ttcctttacg
     5701 gttttcagaa tatccagttc ctgtggatcc agaggaggag tatgtgtgaa ggagtcaccc
     5761 ctaaatgcca ccggcaggat gtggagatcg ccactgatgg aggtgcagtt tttgaagtgt
     5821 ttaatattcg tagcatttat ggagagtgag tctttaaatt caccaatacc tattccgtta
     5881 cacactttgc ggcaaggccc ttcgcacttc ttacacttgc ggacgccgtc ttcctccatc
     5941 tcatagctgt cggccccaca ggctcggacg cacgagccgt gatctgtcac cacataatta
     6001 cggggacact tcttcacgca ggtggcacca aagctgtatt tgccctcggg gttcacatcc
     6061 atctggtacg tggtggggtt gtagagcatg agtggggggc aggtgtcctt gcacgtggct
     6121 tcgtctcgga atttgcggca gaccaggcag tcgctctccc gggggcctgt gcagcctgca
     6181 gcacactggt tgtggcagca gtcactgggg gacttgccac ggcagcgccc ggagcactgc
     6241 tgggcacaga tgattttggt cagtttctgg cagttctcct ctcctgcacc ccagcagctc
     6301 ccattgggac agcttggatc acacttttgg cagctgccca ggtggttctg gaagtccatc
     6361 gacatgttgc tgagaaagtc actgctgact atgtcccgcc actggatgct ctccacgttg
     6421 cacagggcag ggttgttgct gaaccgcacg gcgccatgca ggatttcctg taaatttctc
     6481 atgggcagct ccttcagtcc ggttttattt gcatcatagt tagataagac tgctaaggca
     6541 taggaatttt cgtagtacat atttcctctg atgatctgca ggttttccaa aggaattcgc
     6601 tccactgtgt tgagggcaat gaggacataa ccagccacct cctggatggt ctttaagaag
     6661 gaaagatcat aattcctctg cacataggta atttccaaat tcccaaggac cacctcacag
     6721 ttattgaaca tcctctggag gctgagaaaa tgatcttcaa aagtgcccaa ctgcgtgagc
     6781 ttgttactcg tgccttggca aactttcttt tcctccagag cccgactcgc cgggcagagc
     6841 gcagccagca gcgccaggag cgctgccccg gccgtcccgg agggtcgcat ggcgatcgct
     6901 agccctatag tgagtcgtat taatactcta gccttaagag ctgtaattga actgggagtg
     6961 gacacctgtg gagagaaagg caaagtggat gtcagtaaga ccaataggtg cctatcagaa
     7021 acgcaagagt cttctctgtc tcgacaagcc cagtttctat tggtctcctt aaacctgtct
     7081 tgtaaccttg atacttacct gcccagtgcc tcacgaccaa cttctgcagc ttaagttcga
     7141 gactgttgtg tcagaagcac tgactgcgtt agcaatttaa ctgtgataaa ctaccgcaat
     7201 aaagcttcta gtgatctgac ggttcactaa accagctctg cttatataga cctcccaccg
     7261 tacacgccta ccgcccattt gcgtcaacgg ggcggggtta ttacgacatt ttggaaagtc
     7321 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     7381 tccccgtgag tcaaaccgct atccacgccc attggtgtac tgccaaaacc gcatcaccat
     7441 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc gtaaggtcat
     7501 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcggact
     7561 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     7621 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     7681 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     7741 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     7801 agtcaataat caatgccaac atggcggtca tattggacat gagccaatat aaatgtacat
     7861 attatgatat agatacaacg tatgcaatgg ccaatagcca atattgattt atgctatata
     7921 accaatgaat aatatggcta atggccaata ttgaagatct cgatcctcta cgccggacgc
     7981 atcgtggccg gagtactgcg gtttatctcg atggctacga gggcagacag taagtggatt
     8041 taccataatc ccttaattgt acgcaccgct aaaacgcgtt cagcgcgatc acggcagcag
     8101 acaggtaaaa atggcaacaa accacccgaa aaactgccgc aatcgcgcct gataaatttt
     8161 aaccgtatga atacctatgc aaccagaggg tacaggccac attaccccca cttaatccac
     8221 tgaagctgcc atttttcatg gtttcaccat cccagcgaag ggccatccag cgtgcgttcc
     8281 tgtatttccg gactataaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8401 atcttc
//
