LOCUS 40924_745 8432 bp DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression of epidermal growth factor receptor (EGFR) fused to EGFP . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8432) AUTHORS Carter RE, Sorkin A TITLE Endocytosis of functional epidermal growth factor receptor-green fluorescent protein chimera. JOURNAL J Biol Chem. 1998 Dec 25;273(52):35000-7. PUBMED 9857032 REFERENCE 2 (bases 1 to 8432) TITLE Direct Submission REFERENCE 3 (bases 1 to 8432) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Chem."; date: "1998-12-25"; volume: "273(52)"; pages: "35000-7" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..8432 /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" CDS 725..4354 /codon_start=1 /label=EGFR /note="precursor of human epidermal growth factor receptor" /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA" CDS 4385..5101 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" polyA_signal 5227..5348 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(5355..5810) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 5837..5941 /label=AmpR promoter promoter 5943..6300 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 6335..7126 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7317..7336) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7361..7408 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7737..8325 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" 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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct 4441 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac 4501 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc 4561 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat 4621 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat 4681 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac 4741 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg 4801 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa 4861 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct 4921 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa 4981 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat 5041 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa 5101 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt 5161 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 5221 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 5281 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 5341 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 5401 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 5461 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 5521 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 5581 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 5641 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 5701 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 5761 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac 5821 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 5881 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 5941 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 6001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 6061 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 6121 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6181 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6241 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 6301 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 6361 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 6421 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 6481 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg 6541 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 6601 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 6661 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 6721 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 6781 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 6841 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg 6901 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 6961 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 7021 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 7081 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 7141 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 7201 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 7261 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa 7321 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 7381 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 7441 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 7501 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 7561 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt 7621 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 7681 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 7741 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 7801 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 7861 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 7921 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 7981 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 8041 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 8101 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 8161 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 8221 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 8281 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 8341 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 8401 tcccctgatt ctgtggataa ccgtattacc gc //