LOCUS        EGFR-mEGFP     7314 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Number and brightness analysis of EGFR oligomerization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7314)
  AUTHORS   Gupta A, Lu D, Balasubramanian H, Chi Z, Wohland T
  TITLE     Heptanol-mediated phase separation determines phase preference of 
            molecules in live cell membranes.
  JOURNAL   J Lipid Res. 2022 Jun;63(6):100220. doi: 10.1016/j.jlr.2022.100220. 
            Epub 2022 Apr 28.
  PUBMED    35490741
REFERENCE   2  (bases 1 to 7314)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Lipid 
            Res."; date: "2022-06"; volume: "63(6):100220. doi"; pages: " 
            10.1016/j.jlr.2022.100220. Epub 2022 Apr 28"
COMMENT     Sequence Label: EGFR-mEGFP
FEATURES             Location/Qualifiers
     source          1..7314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(623..694,695..2557,2558..2626,2627..4252)
                     /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"
     CDS             join(4271..4273,4274..4276,4277..4987)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4316..4337)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4577..4596)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4924..4945
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5018..5042)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5071..5192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5082..5101)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5136..5155
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5308..5325)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5479..6067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5559..5578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6238..7029,7030..7098))
                     /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"
     promoter        complement(7099..7203)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7271..7289
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caactttgta tagaaaagtt gtagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc accggtgcca ccatgcgacc ctccgggacg gccggggcag cgctcctggc
      661 gctgctggct gcgctctgcc cggcgagtcg ggctctggag gaaaagaaag tttgccaagg
      721 cacgagtaac aagctcacgc agttgggcac ttttgaagat cattttctca gcctccagag
      781 gatgttcaat aactgtgagg tggtccttgg gaatttggaa attacctatg tgcagaggaa
      841 ttatgatctt tccttcttaa agaccatcca ggaggtggct ggttatgtcc tcattgccct
      901 caacacagtg gagcgaattc ctttggaaaa cctgcagatc atcagaggaa atatgtacta
      961 cgaaaattcc tatgccttag cagtcttatc taactatgat gcaaataaaa ccggactgaa
     1021 ggagctgccc atgagaaatt tacaggaaat cctgcatggc gccgtgcggt tcagcaacaa
     1081 ccctgccctg tgcaacgtgg agagcatcca gtggcgggac atagtcagca gtgactttct
     1141 cagcaacatg tcgatggact tccagaacca cctgggcagc tgccaaaagt gtgatccaag
     1201 ctgtcccaat gggagctgct ggggtgcagg agaggagaac tgccagaaac tgaccaaaat
     1261 catctgtgcc cagcagtgct ccgggcgctg ccgtggcaag tcccccagtg actgctgcca
     1321 caaccagtgt gctgcaggct gcacaggccc ccgggagagc gactgcctgg tctgccgcaa
     1381 attccgagac gaagccacgt gcaaggacac ctgcccccca ctcatgctct acaaccccac
     1441 cacgtaccag atggatgtga accccgaggg caaatacagc tttggtgcca cctgcgtgaa
     1501 gaagtgtccc cgtaattatg tggtgacaga tcacggctcg tgcgtccgag cctgtggggc
     1561 cgacagctat gagatggagg aagacggcgt ccgcaagtgt aagaagtgcg aagggccttg
     1621 ccgcaaagtg tgtaacggaa taggtattgg tgaatttaaa gactcactct ccataaatgc
     1681 tacgaatatt aaacacttca aaaactgcac ctccatcagt ggcgatctcc acatcctgcc
     1741 ggtggcattt aggggtgact ccttcacaca tactcctcct ctggatccac aggaactgga
     1801 tattctgaaa accgtaaagg aaatcacagg gtttttgctg attcaggctt ggcctgaaaa
     1861 caggacggac ctccatgcct ttgagaacct agaaatcata cgcggcagga ccaagcaaca
     1921 tggtcagttt tctcttgcag tcgtcagcct gaacataaca tccttgggat tacgctccct
     1981 caaggagata agtgatggag atgtgataat ttcaggaaac aaaaatttgt gctatgcaaa
     2041 tacaataaac tggaaaaaac tgtttgggac ctccggtcag aaaaccaaaa ttataagcaa
     2101 cagaggtgaa aacagctgca aggccacagg ccaggtctgc catgccttgt gctcccccga
     2161 gggctgctgg ggcccggagc ccagggactg cgtctcttgc cggaatgtca gccgaggcag
     2221 ggaatgcgtg gacaagtgca accttctgga gggtgagcca agggagtttg tggagaactc
     2281 tgagtgcata cagtgccacc cagagtgcct gcctcaggcc atgaacatca cctgcacagg
     2341 acggggacca gacaactgta tccagtgtgc ccactacatt gacggccccc actgcgtcaa
     2401 gacctgcccg gcaggagtca tgggagaaaa caacaccctg gtctggaagt acgcagacgc
     2461 cggccatgtg tgccacctgt gccatccaaa ctgcacctac ggatgcactg ggccaggtct
     2521 tgaaggctgt ccaacgaatg ggcctaagat cccgtccatc gccactggga tggtgggggc
     2581 cctcctcttg ctgctggtgg tggccctggg gatcggcctc ttcatgcgaa ggcgccacat
     2641 cgttcggaag cgcacgctgc ggaggctgct gcaggagagg gagcttgtgg agcctcttac
     2701 acccagtgga gaagctccca accaagctct cttgaggatc ttgaaggaaa ctgaattcaa
     2761 aaagatcaaa gtgctgggct ccggtgcgtt cggcacggtg tataagggac tctggatccc
     2821 agaaggtgag aaagttaaaa ttcccgtcgc tatcaaggaa ttaagagaag caacatctcc
     2881 gaaagccaac aaggaaatcc tcgatgaagc ctacgtgatg gccagcgtgg acaaccccca
     2941 cgtgtgccgc ctgctgggca tctgcctcac ctccaccgtg caactcatca cgcagctcat
     3001 gcccttcggc tgcctcctgg actatgtccg ggaacacaaa gacaatattg gctcccagta
     3061 cctgctcaac tggtgtgtgc agatcgcaaa gggcatgaac tacttggagg accgtcgctt
     3121 ggtgcaccgc gacctggcag ccaggaacgt actggtgaaa acaccgcagc atgtcaagat
     3181 cacagatttt gggctggcca aactgctggg tgcggaagag aaagaatacc atgcagaagg
     3241 aggcaaagtg cctatcaagt ggatggcatt ggaatcaatt ttacacagaa tctataccca
     3301 ccagagtgat gtctggagct acggggtgac cgtttgggag ttgatgacct ttggatccaa
     3361 gccatatgac ggaatccctg ccagcgagat ctcctccatc ctggagaaag gagaacgcct
     3421 ccctcagcca cccatatgta ccatcgatgt ctacatgatc atggtcaagt gctggatgat
     3481 agacgcagat agtcgcccaa agttccgtga gttgatcatc gaattctcca aaatggcccg
     3541 agacccccag cgctaccttg tcattcaggg ggatgaaaga atgcatttgc caagtcctac
     3601 agactccaac ttctaccgtg ccctgatgga tgaagaagac atggacgacg tggtggatgc
     3661 cgacgagtac ctcatcccac agcagggctt cttcagcagc ccctccacgt cacggactcc
     3721 cctcctgagc tctctgagtg caaccagcaa caattccacc gtggcttgca ttgatagaaa
     3781 tgggctgcaa agctgtccca tcaaggaaga cagcttcttg cagcgataca gctcagaccc
     3841 cacaggcgcc ttgactgagg acagcataga cgacaccttc ctcccagtgc ctgaatacat
     3901 aaaccagtcc gttcccaaaa ggcccgctgg ctctgtgcag aatcctgtct atcacaatca
     3961 gcctctgaac cccgcgccca gcagagaccc acactaccag gacccccaca gcactgcagt
     4021 gggcaacccc gagtatctca acactgtcca gcccacctgt gtcaacagca cattcgacag
     4081 ccctgcccac tgggcccaga aaggcagcca ccaaattagc ctggacaacc ctgactacca
     4141 gcaggacttc tttcccaagg aagccaagcc aaatggcatc tttaagggct ccacagctga
     4201 aaatgcagaa tacctaaggg tcgcgccaca aagcagtgaa tttattggag caggaggtgg
     4261 cgggactagt atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4321 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     4381 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4441 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4501 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     4561 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     4621 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     4681 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     4741 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     4801 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4861 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     4921 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     4981 gtacaagggt ggcggagggt ccgctagcta aaagcttacc cagctttctt gtacaaagtg
     5041 gtgatggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     5101 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     5161 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     5221 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     5281 gtagcggccg cggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5341 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5401 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5641 ctttctctct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5881 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6001 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6121 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6181 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6241 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6301 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6361 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6421 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6481 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6541 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6601 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6661 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6721 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6781 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6841 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6901 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6961 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7021 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7081 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7141 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7201 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7261 aacctataaa aataggcgta tcacgaggcc ctttcgtcgg cgcgccgcgg ccgc
//