LOCUS       sequence-478468-        7929 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express EGFR-HiBiT 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 7929)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7929)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: EGFR-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1065..1136,1137..2999,3000..3068,3069..4694)
                     /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(4872..4890)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4927..5048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4938..4957)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4992..5011
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5293..5650
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5293..5484
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5316..5336)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5501..5636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5665..5712
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5744..6538
                     /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(5798..5817)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6408..6427
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6602..6650
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(6889..7477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6969..6988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7593..7876
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatgcga ccctccggga
     1081 cggccggggc agcgctcctg gcgctgctgg ctgcgctctg cccggcgagt cgggctctgg
     1141 aggaaaagaa agtttgccaa ggcacgagta acaagctcac gcagttgggc acttttgaag
     1201 atcattttct cagcctccag aggatgttca ataactgtga ggtggtcctt gggaatttgg
     1261 aaattaccta tgtgcagagg aattatgatc tttccttctt aaagaccatc caggaggtgg
     1321 ctggttatgt cctcattgcc ctcaacacag tggagcgaat tcctttggaa aacctgcaga
     1381 tcatcagagg aaatatgtac tacgaaaatt cctatgcctt agcagtctta tctaactatg
     1441 atgcaaataa aaccggactg aaggagctgc ccatgagaaa tttacaggaa atcctgcatg
     1501 gcgccgtgcg gttcagcaac aaccctgccc tgtgcaacgt ggagagcatc cagtggcggg
     1561 acatagtcag cagtgacttt ctcagcaaca tgtcgatgga cttccagaac cacctgggca
     1621 gctgccaaaa gtgtgatcca agctgtccca atgggagctg ctggggtgca ggagaggaga
     1681 actgccagaa actgaccaaa atcatctgtg cccagcagtg ctccgggcgc tgccgtggca
     1741 agtcccccag tgactgctgc cacaaccagt gtgctgcagg ctgcacaggc ccccgggaga
     1801 gcgactgcct ggtctgccgc aaattccgag acgaagccac gtgcaaggac acctgccccc
     1861 cactcatgct ctacaacccc accacgtacc agatggatgt gaaccccgag ggcaaataca
     1921 gctttggtgc cacctgcgtg aagaagtgtc cccgtaatta tgtggtgaca gatcacggct
     1981 cgtgcgtccg agcctgtggg gccgacagct atgagatgga ggaagacggc gtccgcaagt
     2041 gtaagaagtg cgaagggcct tgccgcaaag tgtgtaacgg aataggtatt ggtgaattta
     2101 aagactcact ctccataaat gctacgaata ttaaacactt caaaaactgc acctccatca
     2161 gtggcgatct ccacatcctg ccggtggcat ttaggggtga ctccttcaca catactcctc
     2221 ctctggatcc acaggaactg gatattctga aaaccgtaaa ggaaatcaca gggtttttgc
     2281 tgattcaggc ttggcctgaa aacaggacgg acctccatgc ctttgagaac ctagaaatca
     2341 tacgcggcag gaccaagcaa catggtcagt tttctcttgc agtcgtcagc ctgaacataa
     2401 catccttggg attacgctcc ctcaaggaga taagtgatgg agatgtgata atttcaggaa
     2461 acaaaaattt gtgctatgca aatacaataa actggaaaaa actgtttggg acctccggtc
     2521 agaaaaccaa aattataagc aacagaggtg aaaacagctg caaggccaca ggccaggtct
     2581 gccatgcctt gtgctccccc gagggctgct ggggcccgga gcccagggac tgcgtctctt
     2641 gccggaatgt cagccgaggc agggaatgcg tggacaagtg caaccttctg gagggtgagc
     2701 caagggagtt tgtggagaac tctgagtgca tacagtgcca cccagagtgc ctgcctcagg
     2761 ccatgaacat cacctgcaca ggacggggac cagacaactg tatccagtgt gcccactaca
     2821 ttgacggccc ccactgcgtc aagacctgcc cggcaggagt catgggagaa aacaacaccc
     2881 tggtctggaa gtacgcagac gccggccatg tgtgccacct gtgccatcca aactgcacct
     2941 acggatgcac agggccaggt cttgaaggct gtccaacgaa tgggcctaag atcccgtcca
     3001 tcgccactgg gatggtgggg gccctcctct tgctgctggt ggtggccctg gggatcggcc
     3061 tcttcatgcg aaggcgccac atcgttcgga agcgcacgct gcggaggctg ctgcaggaga
     3121 gggagcttgt ggagcctctt acacccagtg gagaagctcc caaccaagct ctcttgagga
     3181 tcttgaagga aactgaattc aaaaagatca aagtgctggg ctccggtgcg ttcggcacgg
     3241 tgtataaggg actctggatc ccagaaggtg agaaagttaa aattcccgtc gctatcaagg
     3301 aattaagaga agcaacatct ccgaaagcca acaaggaaat cctcgatgaa gcctacgtga
     3361 tggccagcgt ggacaacccc cacgtgtgcc gcctgctggg catctgcctc acctccaccg
     3421 tgcagctcat cacgcagctc atgcccttcg gctgcctcct ggactatgtc cgggaacaca
     3481 aagacaatat tggctcccag tacctgctca actggtgtgt gcagatcgca aagggcatga
     3541 actacttgga ggaccgtcgc ttggtgcacc gcgacctggc agccaggaac gtactggtga
     3601 aaacaccgca gcatgtcaag atcacagatt ttgggctggc caaactgctg ggtgcggaag
     3661 agaaagaata ccatgcagaa ggaggcaaag tgcctatcaa gtggatggca ttggaatcaa
     3721 ttttacacag aatctatacc caccagagtg atgtctggag ctacggggtg accgtttggg
     3781 agttgatgac ctttggatcc aagccatatg acggaatccc tgccagcgag atctcctcca
     3841 tcctggagaa aggagaacgc ctccctcagc cacccatatg taccatcgat gtctacatga
     3901 tcatggtcaa gtgctggatg atagacgcag atagtcgccc aaagttccgt gagttgatca
     3961 tcgaattctc caaaatggcc cgagaccccc agcgctacct tgtcattcag ggggatgaaa
     4021 gaatgcattt gccaagtcct acagactcca acttctaccg tgccctgatg gatgaagaag
     4081 acatggacga cgtggtggat gccgacgagt acctcatccc acagcagggc ttcttcagca
     4141 gcccctccac gtcacggact cccctcctga gctctctgag tgcaaccagc aacaattcca
     4201 ccgtggcttg cattgataga aatgggctgc aaagctgtcc catcaaggaa gacagcttct
     4261 tgcagcgata cagctcagac cccacaggcg ccttgactga ggacagcata gacgacacct
     4321 tcctcccagt gcctgaatac ataaaccagt ccgttcccaa aaggcccgct ggctctgtgc
     4381 agaatcctgt ctatcacaat cagcctctga accccgcgcc cagcagagac ccacactacc
     4441 aggaccccca cagcactgca gtgggcaacc ccgagtatct caacactgtc cagcccacct
     4501 gtgtcaacag cacattcgac agccctgccc actgggccca gaaaggcagc caccaaatta
     4561 gcctggacaa ccctgactac cagcaggact tctttcccaa ggaagccaag ccaaatggca
     4621 tctttaaggg ctccacagct gaaaatgcag aatacctaag ggtcgcgcca caaagcagtg
     4681 aatttattgg agcagtttct caaggcagtt caggtggtgg cgggagcgga ggtggaggct
     4741 cgagcggtgt gagcggctgg cggctgttca agaagattag ctaaggccgc gactctagag
     4801 tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     4861 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag
     4921 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     4981 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     5041 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     5101 aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcgaattt taacaaaata
     5161 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5221 accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     5281 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     5341 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5401 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5461 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5521 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5581 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5641 cttttgcaaa aagcttaatt aactgttgac aattaatcat cggcatagta tatcggcata
     5701 gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg
     5761 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5821 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5881 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5941 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6001 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6061 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6121 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6181 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6241 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6301 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6361 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6421 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6481 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6541 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     6601 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     6661 gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     6721 catccggggt cagcaccgtt tctgcggact ggctttctac ccggtatcag ctcactcaaa
     6781 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6841 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6901 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6961 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7021 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7081 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7141 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7201 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7261 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7321 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7381 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7441 caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac
     7501 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7561 aaaaaggatc ttcacctaga tccttttata gtccggaaat acaggaacgc acgctggatg
     7621 gcccttcgct gggatggtga aaccatgaaa aatggcagct tcagtggatt aagtgggggt
     7681 aatgtggcct gtaccctctg gttgcatagg tattcatacg gttaaaattt atcaggcgcg
     7741 attgcggcag tttttcgggt ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg
     7801 ctgaacgcgt tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg
     7861 ccctcgtagc catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga
     7921 tcgagatct
//
