LOCUS       sequence-367576-        8155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGF receptor (EGFR) labelled with near-infrared fluorescent protein 
            miRFP670nano3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8155)
  AUTHORS   Oliinyk OS, Baloban M, Clark CL, Carey E, Pletnev S, Nimmerjahn A, 
            Verkhusha VV
  TITLE     Single-domain near-infrared protein provides a scaffold for 
            antigen-dependent fluorescent nanobodies.
  JOURNAL   Nat Methods. 2022 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6.
  PUBMED    35606446
REFERENCE   2  (bases 1 to 8155)
  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: "Nat 
            Methods. 2022 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6."
COMMENT     Sequence Label: pEGFR-miRFP670nano3
FEATURES             Location/Qualifiers
     source          1..8155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(725..796,797..2659,2660..2728,2729..4354)
                     /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"
     regulatory      4379..4388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4950..5071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4987..5006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5041..5060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5078..5533)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5215..5236)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5427..5446
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5560..5664
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5666..6023
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5666..5857
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5689..5709)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5874..6009
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6058..6852
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6058..6852
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6112..6131)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6722..6741
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7040..7059)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7084..7131
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7460..8048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7949..7968
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agagagctta tcgattctag ccgagtcccc gcctcgccgc caacgccaca accaccgcgc
      661 acggccccct gactccgtcc agtattgatc gggagagccg gagcgagctc ttcggggagc
      721 agcgatgcga ccctccggga cggccggggc agcgctcctg gcgctgctgg ctgcgctctg
      781 cccggcgagt cgggctctgg aggaaaagaa agtttgccaa ggcacgagta acaagctcac
      841 gcagttgggc acttttgaag atcattttct cagcctccag aggatgttca ataactgtga
      901 ggtggtcctt gggaatttgg aaattaccta tgtgcagagg aattatgatc tttccttctt
      961 aaagaccatc caggaggtgg ctggttatgt cctcattgcc ctcaacacag tggagcgaat
     1021 tcctttggaa aacctgcaga tcatcagagg aaatatgtac tacgaaaatt cctatgcctt
     1081 agcagtctta tctaactatg atgcaaataa aaccggactg aaggagctgc ccatgagaaa
     1141 tttacaggaa atcctgcatg gcgccgtgcg gttcagcaac aaccctgccc tgtgcaacgt
     1201 ggagagcatc cagtggcggg acatagtcag cagtgacttt ctcagcaaca tgtcgatgga
     1261 cttccagaac cacctgggca gctgccaaaa gtgtgatcca agctgtccca atgggagctg
     1321 ctggggtgca ggagaggaga actgccagaa actgaccaaa atcatctgtg cccagcagtg
     1381 ctccgggcgc tgccgtggca agtcccccag tgactgctgc cacaaccagt gtgctgcagg
     1441 ctgcacaggc ccccgggaga gcgactgcct ggtctgccgc aaattccgag acgaagccac
     1501 gtgcaaggac acctgccccc cactcatgct ctacaacccc accacgtacc agatggatgt
     1561 gaaccccgag ggcaaataca gctttggtgc cacctgcgtg aagaagtgtc cccgtaatta
     1621 tgtggtgaca gatcacggct cgtgcgtccg agcctgtggg gccgacagct atgagatgga
     1681 ggaagacggc gtccgcaagt gtaagaagtg cgaagggcct tgccgcaaag tgtgtaacgg
     1741 aataggtatt ggtgaattta aagactcact ctccataaat gctacgaata ttaaacactt
     1801 caaaaactgc acctccatca gtggcgatct ccacatcctg ccggtggcat ttaggggtga
     1861 ctccttcaca catactcctc ctctggatcc acaggaactg gatattctga aaaccgtaaa
     1921 ggaaatcaca gggtttttgc tgattcaggc ttggcctgaa aacaggacgg acctccatgc
     1981 ctttgagaac ctagaaatca tacgcggcag gaccaagcaa catggtcagt tttctcttgc
     2041 agtcgtcagc ctgaacataa catccttggg attacgctcc ctcaaggaga taagtgatgg
     2101 agatgtgata atttcaggaa acaaaaattt gtgctatgca aatacaataa actggaaaaa
     2161 actgtttggg acctccggtc agaaaaccaa aattataagc aacagaggtg aaaacagctg
     2221 caaggccaca ggccaggtct gccatgcctt gtgctccccc gagggctgct ggggcccgga
     2281 gcccagggac tgcgtctctt gccggaatgt cagccgaggc agggaatgcg tggacaagtg
     2341 caaccttctg gagggtgagc caagggagtt tgtggagaac tctgagtgca tacagtgcca
     2401 cccagagtgc ctgcctcagg ccatgaacat cacctgcaca ggacggggac cagacaactg
     2461 tatccagtgt gcccactaca ttgacggccc ccactgcgtc aagacctgcc cggcaggagt
     2521 catgggagaa aacaacaccc tggtctggaa gtacgcagac gccggccatg tgtgccacct
     2581 gtgccatcca aactgcacct acggatgcac tgggccaggt cttgaaggct gtccaacgaa
     2641 tgggcctaag atcccgtcca tcgccactgg gatggtgggg gccctcctct tgctgctggt
     2701 ggtggccctg gggatcggcc tcttcatgcg aaggcgccac atcgttcgga agcgcacgct
     2761 gcggaggctg ctgcaggaga gggagcttgt ggagcctctt acacccagtg gagaagctcc
     2821 caaccaagct ctcttgagga tcttgaagga aactgaattc aaaaagatca aagtgctggg
     2881 ctccggtgcg ttcggcacgg tgtataaggg actctggatc ccagaaggtg agaaagttaa
     2941 aattcccgtc gctatcaagg aattaagaga agcaacatct ccgaaagcca acaaggaaat
     3001 cctcgatgaa gcctacgtga tggccagcgt ggacaacccc cacgtgtgcc gcctgctggg
     3061 catctgcctc acctccaccg tgcaactcat cacgcagctc atgcccttcg gctgcctcct
     3121 ggactatgtc cgggaacaca aagacaatat tggctcccag tacctgctca actggtgtgt
     3181 gcagatcgca aagggcatga actacttgga ggaccgtcgc ttggtgcacc gcgacctggc
     3241 agccaggaac gtactggtga aaacaccgca gcatgtcaag atcacagatt ttgggctggc
     3301 caaactgctg ggtgcggaag agaaagaata ccatgcagaa ggaggcaaag tgcctatcaa
     3361 gtggatggca ttggaatcaa ttttacacag aatctatacc caccagagtg atgtctggag
     3421 ctacggggtg accgtttggg agttgatgac ctttggatcc aagccatatg acggaatccc
     3481 tgccagcgag atctcctcca tcctggagaa aggagaacgc ctccctcagc cacccatatg
     3541 taccatcgat gtctacatga tcatggtcaa gtgctggatg atagacgcag atagtcgccc
     3601 aaagttccgt gagttgatca tcgaattctc caaaatggcc cgagaccccc agcgctacct
     3661 tgtcattcag ggggatgaaa gaatgcattt gccaagtcct acagactcca acttctaccg
     3721 tgccctgatg gatgaagaag acatggacga cgtggtggat gccgacgagt acctcatccc
     3781 acagcagggc ttcttcagca gcccctccac gtcacggact cccctcctga gctctctgag
     3841 tgcaaccagc aacaattcca ccgtggcttg cattgataga aatgggctgc aaagctgtcc
     3901 catcaaggaa gacagcttct tgcagcgata cagctcagac cccacaggcg ccttgactga
     3961 ggacagcata gacgacacct tcctcccagt gcctgaatac ataaaccagt ccgttcccaa
     4021 aaggcccgct ggctctgtgc agaatcctgt ctatcacaat cagcctctga accccgcgcc
     4081 cagcagagac ccacactacc aggaccccca cagcactgca gtgggcaacc ccgagtatct
     4141 caacactgtc cagcccacct gtgtcaacag cacattcgac agccctgccc actgggccca
     4201 gaaaggcagc caccaaatta gcctggacaa ccctgactac cagcaggact tctttcccaa
     4261 ggaagccaag ccaaatggca tctttaaggg ctccacagct gaaaatgcag aatacctaag
     4321 ggtcgcgcca caaagcagtg aatttattgg agcaccgcgg gcccgggatc caccggtcgc
     4381 caccatggca aacctggaca agatgctgaa caccaccgtg accgaggtgc gcaagttcct
     4441 gcaagcggac agagtgtgcg tgttcaagtt cgaggaagat tactccggca ccgtctcgca
     4501 cgaagccgtg gacgacagat ggattagcat cctgaagacc caggtgcagg acagatactt
     4561 catggaaacc agaggcgagg aatacgtcca cggcagatac caggccatcg ccgacatcta
     4621 cacagccaat ctggtcgagt gctacagaga cctgctgatc gagtttcagg tgcgggccat
     4681 tctggctgtc cccatcctgc aaggcaagaa gctgtggggc ctgctggtgg cccaccaact
     4741 ggccggccct cgggagtggc agacctggga aatcgacttc ctgaaacagc aagccgtggt
     4801 gatgggcatc gccatccagc agagctaggc ggccgcgact ctagatcata atcagccata
     4861 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4921 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4981 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5041 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     5101 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5161 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5221 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5281 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5341 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5401 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5461 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5521 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5581 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5641 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     5701 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5761 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5821 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5881 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5941 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6001 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     6061 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6121 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6181 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     6241 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6301 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6361 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6421 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6481 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6541 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     6601 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6661 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6721 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6781 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6841 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6901 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6961 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     7021 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     7081 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     7141 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     7201 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     7261 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     7321 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7381 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7441 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7501 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7561 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7621 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7681 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7741 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7801 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7861 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7921 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7981 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8041 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8101 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgc
//
