LOCUS Exported 8429 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS perbB1Y1045F-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8429) AUTHORS Offterdinger M, Bastiaens PI TITLE Prolonged EGFR signaling by ERBB2-mediated sequestration at the plasma membrane. JOURNAL Traffic. 2008 Jan;9(1):147-55. Epub 2007 Nov 13. PUBMED 17956594 REFERENCE 2 (bases 1 to 8429) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8429 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 718..4347 /codon_start=1 /product="precursor of human epidermal growth factor receptor" /label=EGFR /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG PEPRDCVSCRNVSRGRECVDKCKLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA CIDRNGLQSCPIKEDSFLQRFSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA" CDS 4375..5094 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4375..5091 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4420..4441) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4681..4700) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5028..5049 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 5217..5338 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5254..5273) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5308..5327 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5345..5800) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5482..5503) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5694..5713 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5827..5931 /gene="bla" /label=AmpR promoter promoter 5933..6290 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5956..5976) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6141..6276 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6203..6222 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6325..7119 /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(6379..6398) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6989..7008 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7307..7326) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7351..7398 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7727..8315 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8216..8235 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagagc 601 ttatcgattc tagccgagtc cccgcctcgc cgccaacgcc acaaccaccg cgcacggccc 661 cctgactccg tccagtattg atcgggagag ccggagcgag ctcttcgggg agcagcgatg 721 cgaccctccg ggacggccgg ggcagcgctc ctggcgctgc tggctgcgct ctgcccggcg 781 agtcgggctc tggaggaaaa gaaagtttgc caaggcacga gtaacaagct cacgcagttg 841 ggcacttttg aagatcattt tctcagcctc cagaggatgt tcaataactg tgaggtggtc 901 cttgggaatt tggaaattac ctatgtgcag aggaattatg atctttcctt cttaaagacc 961 atccaggagg tggctggtta tgtcctcatt gccctcaaca cagtggagcg aattcctttg 1021 gaaaacctgc agatcatcag aggaaatatg tactacgaaa attcctatgc cttagcagtc 1081 ttatctaact atgatgcaaa taaaaccgga ctgaaggagc tgcccatgag aaatttacag 1141 gaaatcctgc atggcgccgt gcggttcagc aacaaccctg ccctgtgcaa cgtggagagc 1201 atccagtggc gggacatagt cagcagtgac tttctcagca acatgtcgat ggacttccag 1261 aaccacctgg gcagctgcca aaagtgtgat ccaagctgtc ccaatgggag ctgctggggt 1321 gcaggagagg agaactgcca gaaactgacc aaaatcatct gtgcccagca gtgctccggg 1381 cgctgccgtg gcaagtcccc cagtgactgc tgccacaacc agtgtgctgc aggctgcaca 1441 ggcccccggg agagcgactg cctggtctgc cgcaaattcc gagacgaagc cacgtgcaag 1501 gacacctgcc ccccactcat gctctacaac cccaccacgt accagatgga tgtgaacccc 1561 gagggcaaat acagctttgg tgccacctgc gtgaagaagt gtccccgtaa ttatgtggtg 1621 acagatcacg gctcgtgcgt ccgagcctgt ggggccgaca gctatgagat ggaggaagac 1681 ggcgtccgca agtgtaagaa gtgcgaaggg ccttgccgca aagtgtgtaa cggaataggt 1741 attggtgaat ttaaagactc actctccata aatgctacga atattaaaca cttcaaaaac 1801 tgcacctcca tcagtggcga tctccacatc ctgccggtgg catttagggg tgactccttc 1861 acacatactc ctcctctgga tccacaggaa ctggatattc tgaaaaccgt aaaggaaatc 1921 acagggtttt tgctgattca ggcttggcct gaaaacagga cggacctcca tgcctttgag 1981 aacctagaaa tcatacgcgg caggaccaag caacatggtc agttttctct tgcagtcgtc 2041 agcctgaaca taacatcctt gggattacgc tccctcaagg agataagtga tggagatgtg 2101 ataatttcag gaaacaaaaa tttgtgctat gcaaatacaa taaactggaa aaaactgttt 2161 gggacctccg gtcagaaaac caaaattata agcaacagag gtgaaaacag ctgcaaggcc 2221 acaggccagg tctgccatgc cttgtgctcc cccgagggct gctggggccc ggagcccagg 2281 gactgcgtct cttgccggaa tgtcagccga ggcagggaat gcgtggacaa gtgcaagctt 2341 ctggagggtg agccaaggga gtttgtggag aactctgagt gcatacagtg ccacccagag 2401 tgcctgcctc aggccatgaa catcacctgc acaggacggg gaccagacaa ctgtatccag 2461 tgtgcccact acattgacgg cccccactgc gtcaagacct gcccggcagg agtcatggga 2521 gaaaacaaca ccctggtctg gaagtacgca gacgccggcc atgtgtgcca cctgtgccat 2581 ccaaactgca cctacggatg cactgggcca ggtcttgaag gctgtccaac gaatgggcct 2641 aagatcccgt ccatcgccac tgggatggtg ggggccctcc tcttgctgct ggtggtggcc 2701 ctggggatcg gcctcttcat gcgaaggcgc cacatcgttc ggaagcgcac gctgcggagg 2761 ctgctgcagg agagggagct tgtggagcct cttacaccca gtggagaagc tcccaaccaa 2821 gctctcttga ggatcttgaa ggaaactgaa ttcaaaaaga tcaaagtgct gggctccggt 2881 gcgttcggca cggtgtataa gggactctgg atcccagaag gtgagaaagt taaaattccc 2941 gtcgctatca aggaattaag agaagcaaca tctccgaaag ccaacaagga aatcctcgat 3001 gaagcctacg tgatggccag cgtggacaac ccccacgtgt gccgcctgct gggcatctgc 3061 ctcacctcca ccgtgcaact catcacgcag ctcatgccct tcggctgcct cctggactat 3121 gtccgggaac acaaagacaa tattggctcc cagtacctgc tcaactggtg tgtgcagatc 3181 gcaaagggca tgaactactt ggaggaccgt cgcttggtgc accgcgacct ggcagccagg 3241 aacgtactgg tgaaaacacc gcagcatgtc aagatcacag attttgggct ggccaaactg 3301 ctgggtgcgg aagagaaaga ataccatgca gaaggaggca aagtgcctat caagtggatg 3361 gcattggaat caattttaca cagaatctat acccaccaga gtgatgtctg gagctacggg 3421 gtgaccgttt gggagttgat gacctttgga tccaagccat atgacggaat ccctgccagc 3481 gagatctcct ccatcctgga gaaaggagaa cgcctccctc agccacccat atgtaccatc 3541 gatgtctaca tgatcatggt caagtgctgg atgatagacg cagatagtcg cccaaagttc 3601 cgtgagttga tcatcgaatt ctccaaaatg gcccgagacc cccagcgcta ccttgtcatt 3661 cagggggatg aaagaatgca tttgccaagt cctacagact ccaacttcta ccgtgccctg 3721 atggatgaag aagacatgga cgacgtggtg gatgccgacg agtacctcat cccacagcag 3781 ggcttcttca gcagcccctc cacgtcacgg actcccctcc tgagctctct gagtgcaacc 3841 agcaacaatt ccaccgtggc ttgcattgat agaaatgggc tgcaaagctg tcccatcaag 3901 gaagacagct tcttgcagcg attcagctca gaccccacag gcgccttgac tgaggacagc 3961 atagacgaca ccttcctccc agtgcctgaa tacataaacc agtccgttcc caaaaggccc 4021 gctggctctg tgcagaatcc tgtctatcac aatcagcctc tgaaccccgc gcccagcaga 4081 gacccacact accaggaccc ccacagcact gcagtgggca accccgagta tctcaacact 4141 gtccagccca cctgtgtcaa cagcacattc gacagccctg cccactgggc ccagaaaggc 4201 agccaccaaa ttagcctgga caaccctgac taccagcagg acttctttcc caaggaagcc 4261 aagccaaatg gcatctttaa gggctccaca gctgaaaatg cagaatacct aagggtcgcg 4321 ccacaaagca gtgaatttat tggagcagcg atcggtggag gtggaggagg caccatggtg 4381 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 4441 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 4501 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 4561 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 4621 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 4681 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 4741 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 4801 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 4861 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 4921 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 4981 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 5041 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc 5101 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa 5161 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact 5221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 5281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag 5341 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc 5401 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc 5461 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac 5521 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca 5581 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg 5641 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag 5701 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc 5761 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga 5821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 5881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg 5941 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 6001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 6061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 6121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 6181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 6241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa 6301 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 6361 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 6421 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 6481 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg 6541 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 6601 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 6661 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 6721 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 6781 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 6841 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 6901 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 6961 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 7021 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 7081 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 7141 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 7201 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 7261 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg 7321 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg 7381 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata 7441 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc 7501 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca 7561 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 7621 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 7681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 7741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 7801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 7861 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 7921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 7981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 8041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 8101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 8161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 8221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 8281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 8341 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 8401 ctgtggataa ccgtattacc gccatgcat //