LOCUS       sequence-479808-        8420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EGFR-LgBiT 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 8420)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8420)
  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            "Unpublished"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-LgBiT, BiBiT-Ready Vector
FEATURES             Location/Qualifiers
     source          1..8420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(806..2431,2432..2500,2501..4363,
                     4364..4435))
                     /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"
     promoter        complement(4474..4492)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4475..4492)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        4612..4915
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5070..5090
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        5123..5170
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5202..5996
                     /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(5256..5275)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5866..5885
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6154..6742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6234..6253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6868..6916
                     /note="synthetic polyadenylation signal"
     CDS             complement(6940..7977)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             6953..6961
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(8008..8365)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8022..8157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8174..8365
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8322..8342
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaactgctc caataaattc actgctttgt ggcgcgaccc
      841 ttaggtattc tgcattttca gctgtggagc ccttaaagat gccatttggc ttggcttcct
      901 tgggaaagaa gtcctgctgg tagtcagggt tgtccaggct aatttggtgg ctgcctttct
      961 gggcccagtg ggcagggctg tcgaatgtgc tgttgacaca ggtgggctgg acagtgttga
     1021 gatactcggg gttgcccact gcagtgctgt gggggtcctg gtagtgtggg tctctgctgg
     1081 gcgcggggtt cagaggctga ttgtgataga caggattctg cacagagcca gcgggccttt
     1141 tgggaacgga ctggtttatg tattcaggca ctgggaggaa ggtgtcgtct atgctgtcct
     1201 cagtcaaggc gcctgtgggg tctgagctgt atcgctgcaa gaagctgtct tccttgatgg
     1261 gacagctttg cagcccattt ctatcaatgc aagccacggt ggaattgttg ctggttgcac
     1321 tcagagagct caggagggga gtccgtgacg tggaggggct gctgaagaag ccctgctgtg
     1381 ggatgaggta ctcgtcggca tccaccacgt cgtccatgtc ttcttcatcc atcagggcac
     1441 ggtagaagtt ggagtctgta ggacttggca aatgcattct ttcatccccc tgaatgacaa
     1501 ggtagcgctg ggggtctcgg gccattttgg agaattcgat gatcaactca cggaactttg
     1561 ggcgactatc tgcgtctatc atccagcact tgaccatgat catgtagaca tcgatggtac
     1621 atatgggtgg ctgagggagg cgttctcctt tctccaggat ggaggagatc tcgctggcag
     1681 ggattccgtc atatggcttg gatccaaagg tcatcaactc ccaaacggtc accccgtagc
     1741 tccagacatc actctggtgg gtatagattc tgtgtaaaat tgattccaat gccatccact
     1801 tgataggcac tttgcctcct tctgcatggt attctttctc ttccgcaccc agcagtttgg
     1861 ccagcccaaa atctgtgatc ttgacatgct gcggtgtttt caccagtacg ttcctggctg
     1921 ccaggtcgcg gtgcaccaag cgacggtcct ccaagtagtt catgcccttt gcgatctgca
     1981 cacaccagtt gagcaggtac tgggagccaa tattgtcttt gtgttcccgg acatagtcca
     2041 ggaggcagcc gaagggcatg agctgcgtga tgagctgcac ggtggaggtg aggcagatgc
     2101 ccagcaggcg gcacacgtgg gggttgtcca cgctggccat cacgtaggct tcatcgagga
     2161 tttccttgtt ggctttcgga gatgttgctt ctcttaattc cttgatagcg acgggaattt
     2221 taactttctc accttctggg atccagagtc ccttatacac cgtgccgaac gcaccggagc
     2281 ccagcacttt gatctttttg aattcagttt ccttcaagat cctcaagaga gcttggttgg
     2341 gagcttctcc actgggtgta agaggctcca caagctccct ctcctgcagc agcctccgca
     2401 gcgtgcgctt ccgaacgatg tggcgccttc gcatgaagag gccgatcccc agggccacca
     2461 ccagcagcaa gaggagggcc cccaccatcc cagtggcgat ggacgggatc ttaggcccat
     2521 tcgttggaca gccttcaaga cctggccctg tgcatccgta ggtgcagttt ggatggcaca
     2581 ggtggcacac atggccggcg tctgcgtact tccagaccag ggtgttgttt tctcccatga
     2641 ctcctgccgg gcaggtcttg acgcagtggg ggccgtcaat gtagtgggca cactggatac
     2701 agttgtctgg tccccgtcct gtgcaggtga tgttcatggc ctgaggcagg cactctgggt
     2761 ggcactgtat gcactcagag ttctccacaa actcccttgg ctcaccctcc agaaggttgc
     2821 acttgtccac gcattccctg cctcggctga cattccggca agagacgcag tccctgggct
     2881 ccgggcccca gcagccctcg ggggagcaca aggcatggca gacctggcct gtggccttgc
     2941 agctgttttc acctctgttg cttataattt tggttttctg accggaggtc ccaaacagtt
     3001 ttttccagtt tattgtattt gcatagcaca aatttttgtt tcctgaaatt atcacatctc
     3061 catcacttat ctccttgagg gagcgtaatc ccaaggatgt tatgttcagg ctgacgactg
     3121 caagagaaaa ctgaccatgt tgcttggtcc tgccgcgtat gatttctagg ttctcaaagg
     3181 catggaggtc cgtcctgttt tcaggccaag cctgaatcag caaaaaccct gtgatttcct
     3241 ttacggtttt cagaatatcc agttcctgtg gatccagagg aggagtatgt gtgaaggagt
     3301 cacccctaaa tgccaccggc aggatgtgga gatcgccact gatggaggtg cagtttttga
     3361 agtgtttaat attcgtagca tttatggaga gtgagtcttt aaattcacca atacctattc
     3421 cgttacacac tttgcggcaa ggcccttcgc acttcttaca cttgcggacg ccgtcttcct
     3481 ccatctcata gctgtcggcc ccacaggctc ggacgcacga gccgtgatct gtcaccacat
     3541 aattacgggg acacttcttc acgcaggtgg caccaaagct gtatttgccc tcggggttca
     3601 catccatctg gtacgtggtg gggttgtaga gcatgagtgg ggggcaggtg tccttgcacg
     3661 tggcttcgtc tcggaatttg cggcagacca ggcagtcgct ctcccggggg cctgtgcagc
     3721 ctgcagcaca ctggttgtgg cagcagtcac tgggggactt gccacggcag cgcccggagc
     3781 actgctgggc acagatgatt ttggtcagtt tctggcagtt ctcctctcct gcaccccagc
     3841 agctcccatt gggacagctt ggatcacact tttggcagct gcccaggtgg ttctggaagt
     3901 ccatcgacat gttgctgaga aagtcactgc tgactatgtc ccgccactgg atgctctcca
     3961 cgttgcacag ggcagggttg ttgctgaacc gcacggcgcc atgcaggatt tcctgtaaat
     4021 ttctcatggg cagctccttc agtccggttt tatttgcatc atagttagat aagactgcta
     4081 aggcatagga attttcgtag tacatatttc ctctgatgat ctgcaggttt tccaaaggaa
     4141 ttcgctccac tgtgttgagg gcaatgagga cataaccagc cacctcctgg atggtcttta
     4201 agaaggaaag atcataattc ctctgcacat aggtaatttc caaattccca aggaccacct
     4261 cacagttatt gaacatcctc tggaggctga gaaaatgatc ttcaaaagtg cccaactgcg
     4321 tgagcttgtt actcgtgcct tggcaaactt tcttttcctc cagagcccga ctcgccgggc
     4381 agagcgcagc cagcagcgcc aggagcgctg ccccggccgt cccggagggt cgcatggcga
     4441 tcgctagctg caaaaagaac aagtagcttg tattctatag tgtcacctaa atgggcgatc
     4501 tgacggttca ctaaaccagc tctgcttata taggcctccc tggcgcgcca ctagttatta
     4561 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     4621 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     4681 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     4741 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     4801 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     4861 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     4921 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     4981 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     5041 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     5101 ggtcaaatcg aattctaata ctgttgacaa ttaatcatcg gcatagtata tcggcatagt
     5161 ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat
     5221 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5281 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5341 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5401 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5461 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5521 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5581 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5641 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5701 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5761 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5821 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5881 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5941 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg
     6001 aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt
     6061 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6721 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6841 ctagatcctt ttatagtccg gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa
     6901 ataaagatat tttattgcgg ccatcgtgat ggctagagtc taaactcgac ctacctcctt
     6961 agcgcgcggt cgtgtactgg gccggcggtt gccgctgtcg gccagcacct cgacgcagcc
     7021 gtcggtccat acggctgcgc tccggcgagc gatttgtgtg cgaccgacgg tgccggcccc
     7081 gctgcggacg atggcatcgc agcggccttg agcccaggca gcatcgtcga agttgccgtc
     7141 cacgaggctc tggtagagct gatccaggcc gatgcgcagc atgtaggctc gcagacgagg
     7201 gctgccggcc agctcgggat gccggcgctc gaagtagcga gtctgctgct ccatgcaagc
     7261 cagccagggc cgccagaaga agatgttggc cacctcgtac tgactgtccc cgaacatagc
     7321 ttcggaccag tcgattacgg cggtgatgcg gccgttgtcg gtcaggacgt tgttgctgcc
     7381 gaagtcggca tggacgaggt ggcgcacctc gggacagtct tcggcccaca gcatcagttc
     7441 gtccagggct tgagctacgc tggcggacac ggtgtcgtcc atcacggtct gccagtggta
     7501 gacatgggga tcagcaatgg cgcaaatgaa atcccgccag gttgtgtact ggccgatgcc
     7561 ttggggcccg aatgggccga agccgctggt ttgactgagg tcggcggcgg caatagcatc
     7621 catagcttcg gcgacaggct gtaacacagc aggcagctct gtttcgggga ggtcttggag
     7681 agtaacgcct tgggcgcgtc tactgatgca gtatgtcagg ctctcgctga actcgccgat
     7741 gtccaacact tcggggatgg gtagtgcagc gctggcgaag tggcggtaca cgtagcggtc
     7801 tttgtagaag ccatcagcgc agctattcac ccgcagtaca tagccgcgtc cgccgacatc
     7861 gaagctgaag gctcggctct cttcgccctc cgacaactgc atcaggtcgc tcacactgtc
     7921 gaacttctcg atgagaaatt tttcaacgct ggtagcggtg agttcgggct tcttcatggt
     7981 ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     8041 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     8101 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     8161 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     8221 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     8281 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     8341 ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat
     8401 ttcaggccat ggtgctgcgc
//
