LOCUS Exported 7891 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for soluble ACE2-Fc fusion (IgG1) high affinity 2.2. ACCESSION . VERSION . KEYWORDS pcDNA3-sACE2v2.2-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" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3248..3265) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3254..3478 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3524..3952 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3611..3630) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3821..3842 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3961..3981) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3966..4295 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4146..4281 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4208..4227 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4362..5156 /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(4416..4435) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5026..5045 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5330..5451 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5367..5386) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5421..5440 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5500..5516) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5500..5516) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5513..5535) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5524..5540 /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(5548..5578) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5593..5614 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5731..5748) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5902..6490) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5982..6001) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6661..7521) /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 7284..7303 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7522..7626) /gene="bla" /label=AmpR promoter primer_bind complement(7701..7720) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcgcca ccatgtcaag ctcttcctgg ctccttctca gccttgttgc tgtaactgct 721 gctcagtcca ccattgagga acaggccaag tactttttgg acaagtttaa ccacgaagcc 781 gaagacctgt tctatcaaag ttcacttgct tcttggaatt ataacaccaa tattactgaa 841 gagaatgtcc aaaacatgaa taatgctggg gacaaatggt ctgccttttt aaaggaacag 901 tccacacttg cccaaatgta tccactacaa gaaattcaga atctcacagt caagcttcag 961 ctgcaggctc ttcagcaaaa tgggtcttca gtgctctcag aagacaagag caaacggttg 1021 aacacaattc taaatacaat gagcaccatc tacagtactg gaaaagtttg taacccagat 1081 aatccacaag aatgcttatt acttgaacca ggtttgaatg aaataatggc aaacagttta 1141 gactacaatg agaggctctg ggcttgggaa agctggagat ctgaggtcgg caagcagctg 1201 aggccattat atgaagagta tgtggtcttg aaaaatgaga tggcaagagc aaatcattat 1261 gaggactatg gggattattg gagaggagac tatgaagtaa atggggtaga tggctatgac 1321 tacagccgcg gccagttgat tgaagatgtg gaacatacct ttgaagagat taaaccatta 1381 tatgaacatc ttcatgccta tgtgagggca aagttgatga atgcctatcc ttcctatatc 1441 agtccaattg gatgcctccc tgctcatttg cttggtgata tgtggggtag attttggaca 1501 aatctgtact ctttgacagt tccctttgga cagaaaccaa acatagatgt tactgatgca 1561 atggtggacc aggcctggga tgcacagaga atattcaagg aggccgagaa gttctttgta 1621 tctgttggtc ttcctaatat gactcaagga ttctgggaat attccatgct aacggaccca 1681 ggaaatgttc agaaagcagt ctgccatccc acagcttggg acctggggaa gggcgacttc 1741 aggattctta tgtgcacaaa ggtgacaatg gacgacttcc tgacagctca tcatgagatg 1801 gggcatatcc agtatgatat ggcatatctt gcacaacctt ttctgctaag aaatggagct 1861 aatgaaggat tccatgaagc tgttggggaa atcatgtcac tttctgcagc cacacctaag 1921 catttaaaat ccattggtct tctgtcaccc gattttcaag aagacaatga aacagaaata 1981 aacttcctgc tcaaacaagc actcacgatt gttgggactc tgccatttac ttacatgtta 2041 gagaagtgga ggtggatggt ctttaaaggg gaaattccca aagaccagtg gatgaaaaag 2101 tggtgggaga tgaagcgaga gatagttggg gtggtggaac ctgtgcccca tgatgaaaca 2161 tactgtgacc ccgcatctct gttccatgtt tctaatgatt actcattcat tcgatattac 2221 acaaggaccc tttaccaatt ccagtttcaa gaagcacttt gtcaagcagc taaacatgaa 2281 ggccctctgc acaaatgtga catctcaaac tctacagaag ctggacagaa actgttcaat 2341 atgctgaggc ttggaaaatc agaaccctgg accctagcat tggaaaatgt tgtaggagca 2401 aagaacatga atgtaaggcc actgctcaac tactttgagc ccttatttac ctggctgaaa 2461 gaccagaaca agaattcttt tgtgggatgg agtaccgact ggagtccata tgcagacgga 2521 tccgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 2581 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 2641 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 2701 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 2761 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 2821 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 2881 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 2941 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 3001 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 3061 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 3121 gggaacgtct tctcatgctc cgtgatgcac gaggctctgc acaaccacta cacgcagaag 3181 agcctctccc tgtctccggg taaatgataa ctcgagtcta gagggcccgt ttaaacccgc 3241 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 3301 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 3361 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 3421 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 3481 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 3541 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3601 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3661 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 3721 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3781 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3841 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 3901 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 3961 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 4021 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 4081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 4141 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 4201 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 4261 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 4321 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 4381 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 4441 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 4501 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 4561 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4621 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4681 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4741 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4801 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4861 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 4921 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4981 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 5041 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 5101 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 5161 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 5221 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 5281 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 5341 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5401 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 5461 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 5521 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 5581 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 5641 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 5701 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 5761 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 5821 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 5881 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 5941 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 6001 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 6061 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 6121 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 6181 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 6241 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 6301 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 6361 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 6421 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6481 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6541 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6601 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 6661 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6721 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6781 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6841 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6901 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6961 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7021 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7081 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7141 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7201 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 7261 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7321 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7381 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7441 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7501 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7561 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 7621 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat 7681 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg 7741 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca 7801 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg 7861 cttcgcgatg tacgggccag atatacgcgt t //