LOCUS       sequence-401142-        7221 bp    DNA     circular SYN 15-DEC-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 7221)
  AUTHORS   Balasubramanian H, Sankaran J, Pandey S, Goh CJH, Wohland T
  TITLE     The dependence of EGFR oligomerization on environment and structure:
            A camera-based N
  JOURNAL   Biophys J. 2022 Dec 6;121(23):4452-4466. doi: 
            10.1016/j.bpj.2022.11.003. Epub 2022 Nov 5.
  PUBMED    36335429
REFERENCE   2  (bases 1 to 7221)
  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"; doi: 
            "10.1016/j.bpj.2022.11.003"; journalName: "Biophys J"; date: 
            "2022-12-6- 6"; volume: "121"; issue: "23"; pages: "4452-4466"
COMMENT     Sequence Label: EGFR∆550-580-mEGFP
FEATURES             Location/Qualifiers
     source          1..7221
                     /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(4178..4180,4181..4183,4184..4894)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYQTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4223..4244)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4831..4852
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4925..4949)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4978..5099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4989..5008)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5043..5062
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5215..5232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5386..5974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5466..5485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6145..6936,6937..7005))
                     /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(7006..7110)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7178..7196
                     /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 agggagtttc agtgtgccca
     2281 ctacattgac ggcccccact gcgtcaagac ctgcccggca ggagtcatgg gagaaaacaa
     2341 caccctggtc tggaagtacg cagacgccgg ccatgtgtgc cacctgtgcc atccaaactg
     2401 cacctacgga tgcactgggc caggtcttga aggctgtcca acgaatgggc ctaagatccc
     2461 gtccatcgcc actgggatgg tgggggccct cctcttgctg ctggtggtgg ccctggggat
     2521 cggcctcttc atgcgaaggc gccacatcgt tcggaagcgc acgctgcgga ggctgctgca
     2581 ggagagggag cttgtggagc ctcttacacc cagtggagaa gctcccaacc aagctctctt
     2641 gaggatcttg aaggaaactg aattcaaaaa gatcaaagtg ctgggctccg gtgcgttcgg
     2701 cacggtgtat aagggactct ggatcccaga aggtgagaaa gttaaaattc ccgtcgctat
     2761 caaggaatta agagaagcaa catctccgaa agccaacaag gaaatcctcg atgaagccta
     2821 cgtgatggcc agcgtggaca acccccacgt gtgccgcctg ctgggcatct gcctcacctc
     2881 caccgtgcaa ctcatcacgc agctcatgcc cttcggctgc ctcctggact atgtccggga
     2941 acacaaagac aatattggct cccagtacct gctcaactgg tgtgtgcaga tcgcaaaggg
     3001 catgaactac ttggaggacc gtcgcttggt gcaccgcgac ctggcagcca ggaacgtact
     3061 ggtgaaaaca ccgcagcatg tcaagatcac agattttggg ctggccaaac tgctgggtgc
     3121 ggaagagaaa gaataccatg cagaaggagg caaagtgcct atcaagtgga tggcattgga
     3181 atcaatttta cacagaatct atacccacca gagtgatgtc tggagctacg gggtgaccgt
     3241 ttgggagttg atgacctttg gatccaagcc atatgacgga atccctgcca gcgagatctc
     3301 ctccatcctg gagaaaggag aacgcctccc tcagccaccc atatgtacca tcgatgtcta
     3361 catgatcatg gtcaagtgct ggatgataga cgcagatagt cgcccaaagt tccgtgagtt
     3421 gatcatcgaa ttctccaaaa tggcccgaga cccccagcgc taccttgtca ttcaggggga
     3481 tgaaagaatg catttgccaa gtcctacaga ctccaacttc taccgtgccc tgatggatga
     3541 agaagacatg gacgacgtgg tggatgccga cgagtacctc atcccacagc agggcttctt
     3601 cagcagcccc tccacgtcac ggactcccct cctgagctct ctgagtgcaa ccagcaacaa
     3661 ttccaccgtg gcttgcattg atagaaatgg gctgcaaagc tgtcccatca aggaagacag
     3721 cttcttgcag cgatacagct cagaccccac aggcgccttg actgaggaca gcatagacga
     3781 caccttcctc ccagtgcctg aatacataaa ccagtccgtt cccaaaaggc ccgctggctc
     3841 tgtgcagaat cctgtctatc acaatcagcc tctgaacccc gcgcccagca gagacccaca
     3901 ctaccaggac ccccacagca ctgcagtggg caaccccgag tatctcaaca ctgtccagcc
     3961 cacctgtgtc aacagcacat tcgacagccc tgcccactgg gcccagaaag gcagccacca
     4021 aattagcctg gacaaccctg actaccagca ggacttcttt cccaaggaag ccaagccaaa
     4081 tggcatcttt aagggctcca cagctgaaaa tgcagaatac ctaagggtcg cgccacaaag
     4141 cagtgaattt attggagcag gaggtggcgg gactagtatg gtgagcaagg gcgaggagct
     4201 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     4261 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     4321 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     4381 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     4441 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacca
     4501 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     4561 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     4621 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     4681 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     4741 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     4801 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     4861 cgggatcact ctcggcatgg acgagctgta caagggtggc ggagggtccg ctagctaaaa
     4921 gcttacccag ctttcttgta caaagtggtg atggccggcc gcttcgagca gacatgataa
     4981 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     5041 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     5101 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     5161 aaagcaagta aaacctctac aaatgtggta gcggccgcgg cgctcttccg cttcctcgct
     5221 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5281 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5521 cctgccgctt accggatacc tgtccgcctt tctctcttcg ggaagcgtgg cgctttctca
     5581 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5701 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5761 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5821 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5881 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5941 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6001 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6061 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7141 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7201 tcgtcggcgc gccgcggccg c
//
