LOCUS Exported 7891 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for soluble ACE2-Fc fusion (IgG1) high affinity 3. ACCESSION . VERSION . KEYWORDS pcDNA3-sACE2v3-Fc(IgG1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7891) AUTHORS Chan KK, Dorosky D, Sharma P, Abbasi SA, Dye JM, Kranz DM, Herbert AS, Procko E TITLE Engineering human ACE2 to optimize binding to the spike protein of SARS coronavirus 2. JOURNAL Science. 2020 Sep 4;369(6508):1261-1265. doi: 10.1126/science.abc0870. Epub 2020 Aug 4. PUBMED 32753553 REFERENCE 2 (bases 1 to 7891) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7891 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..204 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 155..175 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 249..268 /label=T7 /note="T7 promoter, forward primer" promoter 249..267 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2868..2885) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2874..3098 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3144..3572 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3231..3250) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3441..3462 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3581..3601) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3586..3915 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3766..3901 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3828..3847 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3982..4776 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4036..4055) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4646..4665 /label=Neo-F /note="Neomycin resistance gene, forward primer" 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" primer_bind complement(5120..5136) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5120..5136) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5133..5155) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5144..5160 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5168..5198) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5213..5234 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5351..5368) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5522..6110) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5602..5621) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6281..7141) /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" primer_bind 6904..6923 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7142..7246) /gene="bla" /label=AmpR promoter primer_bind complement(7321..7340) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 7512..7891 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt 61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg 181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt 241 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgcca ccatgtcaag 301 ctcttcctgg ctccttctca gccttgttgc tgtaactgct gctcagtcca ccattgagga 361 acaggtcaag tactttttgg acaagtttaa ccacgaagcc gaagacctgt tctatcaaag 421 ttcacttgct tcttggaatt ataacaccaa tattactgaa gagaatgtcc aaaacatgaa 481 taatgctggg gacaaatggt ctgccttttt aaaggaacag tccacacttg cccaaatgta 541 tccactacaa gaaattcaga atctccaagt caagcttcag ctgcaggctc ttcagcaaaa 601 tgggtcttca gtgctctcag aagacaagag caaacggttg aacacaattc taaatacaat 661 gagcaccatc tacagtactg gaaaagtttg taacccagat aatccacaag aatgcttatt 721 acttgaacca ggtttgaatg aaataatggc aaacagttta gactacaatg agaggctctg 781 ggcttgggaa agctggagat ctgaggtcgg caagcagctg aggccattat atgaagagta 841 tgtggtcttg aaaaatgaga tggcaagagc aaatcattat gaggactatg gggattattg 901 gagaggagac tatgaagtaa atggggtaga tggctatgac tacagccgcg gccagttgat 961 tgaagatgtg gaacatacct ttgaagagat taaaccatta tatgaacatc ttcatgccta 1021 tgtgagggca aagttgatga atgcctatcc ttcctatatc agtccaattg gatgcctccc 1081 tgctcatttg cttggtgata tgtggggtag attttggaca aatctgtact ctttgacagt 1141 tccctttgga cagaaaccaa acatagatgt tactgatgca atggtggacc aggcctggga 1201 tgcacagaga atattcaagg aggccgagaa gttctttgta tctgttggtc ttcctaatat 1261 gactccagga ttctgggaaa attccatgct aacggaccca ggaaatgttc agaaagcagt 1321 ctgccatccc acagcttggg acctggggaa gggcgacttc aggattctta tgtgcacaaa 1381 ggtgacaatg gacgacttcc tgacagctca tcatgagatg gggcatatcc agtatgatat 1441 ggcatatctt gcacaacctt ttctgctaag aaatggagct aatgaaggat tccatgaagc 1501 tgttggggaa atcatgtcac tttctgcagc cacacctaag catttaaaat ccattggtct 1561 tctgtcaccc gattttcaag aagacaatga aacagaaata aacttcctgc tcaaacaagc 1621 actcacgatt gttgggactc tgccatttac ttacatgtta gagaagtgga ggtggatggt 1681 ctttaaaggg gaaattccca aagaccagtg gatgaaaaag tggtgggaga tgaagcgaga 1741 gatagttggg gtggtggaac ctgtgcccca tgatgaaaca tactgtgacc ccgcatctct 1801 gttccatgtt tctaatgatt actcattcat tcgatattac acaaggaccc tttaccaatt 1861 ccagtttcaa gaagcacttt gtcaagcagc taaacatgaa ggccctctgc acaaatgtga 1921 catctcaaac tctacagaag ctggacagaa actgttcaat atgctgaggc ttggaaaatc 1981 agaaccctgg accctagcat tggaaaatgt tgtaggagca aagaacatga atgtaaggcc 2041 actgctcaac tactttgagc ccttatttac ctggctgaaa gaccagaaca agaattcttt 2101 tgtgggatgg agtaccgact ggagtccata tgcagacgga tccgacaaaa ctcacacatg 2161 cccaccgtgc ccagcacctg aactcctggg gggaccgtca gtcttcctct tccccccaaa 2221 acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg tggtggacgt 2281 gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg aggtgcataa 2341 tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg tcagcgtcct 2401 caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg tctccaacaa 2461 agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc cccgagaacc 2521 acaggtgtac accctgcccc catcccggga ggagatgacc aagaaccagg tcagcctgac 2581 ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga gcaatgggca 2641 gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct ccttcttcct 2701 ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct tctcatgctc 2761 cgtgatgcac gaggctctgc acaaccacta cacgcagaag agcctctccc tgtctccggg 2821 taaatgataa ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 2881 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 2941 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 3001 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 3061 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 3121 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 3181 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 3241 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 3301 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 3361 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 3421 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 3481 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 3541 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 3601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3661 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3721 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 3781 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 3841 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 3901 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 3961 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 4021 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 4081 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 4141 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 4201 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 4261 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 4321 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 4381 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 4441 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 4501 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 4561 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 4621 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 4681 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 4741 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 4801 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 4861 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 4921 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 4981 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 5041 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 5101 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 5161 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 5221 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 5281 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 5341 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 5401 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 5461 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 5521 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 5581 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5641 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5701 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5761 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 5821 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5881 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5941 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 6001 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 6061 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 6121 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 6181 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 6241 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 6301 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6361 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6421 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6481 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6541 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6601 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6661 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6721 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6781 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6841 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6901 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6961 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 7021 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 7081 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 7141 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 7201 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 7261 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag 7321 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga 7381 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa 7441 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag 7501 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt 7561 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg 7621 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 7681 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 7741 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 7801 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 7861 catctacgta ttagtcatcg ctattaccat g //