LOCUS Exported 7891 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for soluble ACE2-Fc fusion (IgG1). ACCESSION . VERSION . KEYWORDS pcDNA3-sACE2(WT)-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" enhancer 155..534 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 535..738 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 689..709 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 783..802 /label=T7 /note="T7 promoter, forward primer" promoter 783..801 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3402..3419) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3408..3632 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3678..4106 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3765..3784) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3975..3996 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4115..4135) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4120..4449 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4300..4435 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4362..4381 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4516..5310 /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(4570..4589) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5180..5199 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5484..5605 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5521..5540) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5575..5594 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5654..5670) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5654..5670) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5667..5689) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5678..5694 /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(5702..5732) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5747..5768 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5885..5902) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6056..6644) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6136..6155) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6815..7675) /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 7438..7457 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7676..7780) /gene="bla" /label=AmpR promoter primer_bind complement(7855..7874) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca 61 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg 121 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa 181 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa 241 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata 301 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag 361 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc 421 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta 481 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg 541 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt 601 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca 661 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag 721 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa 781 attaatacga ctcactatag ggagacccaa gctggctagc gccaccatgt caagctcttc 841 ctggctcctt ctcagccttg ttgctgtaac tgctgctcag tccaccattg aggaacaggc 901 caagacattt ttggacaagt ttaaccacga agccgaagac ctgttctatc aaagttcact 961 tgcttcttgg aattataaca ccaatattac tgaagagaat gtccaaaaca tgaataatgc 1021 tggggacaaa tggtctgcct ttttaaagga acagtccaca cttgcccaaa tgtatccact 1081 acaagaaatt cagaatctca cagtcaagct tcagctgcag gctcttcagc aaaatgggtc 1141 ttcagtgctc tcagaagaca agagcaaacg gttgaacaca attctaaata caatgagcac 1201 catctacagt actggaaaag tttgtaaccc agataatcca caagaatgct tattacttga 1261 accaggtttg aatgaaataa tggcaaacag tttagactac aatgagaggc tctgggcttg 1321 ggaaagctgg agatctgagg tcggcaagca gctgaggcca ttatatgaag agtatgtggt 1381 cttgaaaaat gagatggcaa gagcaaatca ttatgaggac tatggggatt attggagagg 1441 agactatgaa gtaaatgggg tagatggcta tgactacagc cgcggccagt tgattgaaga 1501 tgtggaacat acctttgaag agattaaacc attatatgaa catcttcatg cctatgtgag 1561 ggcaaagttg atgaatgcct atccttccta tatcagtcca attggatgcc tccctgctca 1621 tttgcttggt gatatgtggg gtagattttg gacaaatctg tactctttga cagttccctt 1681 tggacagaaa ccaaacatag atgttactga tgcaatggtg gaccaggcct gggatgcaca 1741 gagaatattc aaggaggccg agaagttctt tgtatctgtt ggtcttccta atatgactca 1801 aggattctgg gaaaattcca tgctaacgga cccaggaaat gttcagaaag cagtctgcca 1861 tcccacagct tgggacctgg ggaagggcga cttcaggatt cttatgtgca caaaggtgac 1921 aatggacgac ttcctgacag ctcatcatga gatggggcat atccagtatg atatggcata 1981 tgctgcacaa ccttttctgc taagaaatgg agctaatgaa ggattccatg aagctgttgg 2041 ggaaatcatg tcactttctg cagccacacc taagcattta aaatccattg gtcttctgtc 2101 acccgatttt caagaagaca atgaaacaga aataaacttc ctgctcaaac aagcactcac 2161 gattgttggg actctgccat ttacttacat gttagagaag tggaggtgga tggtctttaa 2221 aggggaaatt cccaaagacc agtggatgaa aaagtggtgg gagatgaagc gagagatagt 2281 tggggtggtg gaacctgtgc cccatgatga aacatactgt gaccccgcat ctctgttcca 2341 tgtttctaat gattactcat tcattcgata ttacacaagg accctttacc aattccagtt 2401 tcaagaagca ctttgtcaag cagctaaaca tgaaggccct ctgcacaaat gtgacatctc 2461 aaactctaca gaagctggac agaaactgtt caatatgctg aggcttggaa aatcagaacc 2521 ctggacccta gcattggaaa atgttgtagg agcaaagaac atgaatgtaa ggccactgct 2581 caactacttt gagcccttat ttacctggct gaaagaccag aacaagaatt cttttgtggg 2641 atggagtacc gactggagtc catatgcaga cggatccgac aaaactcaca catgcccacc 2701 gtgcccagca cctgaactcc tggggggacc gtcagtcttc ctcttccccc caaaacccaa 2761 ggacaccctc atgatctccc ggacccctga ggtcacatgc gtggtggtgg acgtgagcca 2821 cgaagaccct gaggtcaagt tcaactggta cgtggacggc gtggaggtgc ataatgccaa 2881 gacaaagccg cgggaggagc agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt 2941 cctgcaccag gactggctga atggcaagga gtacaagtgc aaggtctcca acaaagccct 3001 cccagccccc atcgagaaaa ccatctccaa agccaaaggg cagccccgag aaccacaggt 3061 gtacaccctg cccccatccc gggaggagat gaccaagaac caggtcagcc tgacctgcct 3121 ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga 3181 gaacaactac aagaccacgc ctcccgtgct ggactccgac ggctccttct tcctctacag 3241 caagctcacc gtggacaaga gcaggtggca gcaggggaac gtcttctcat gctccgtgat 3301 gcacgaggct ctgcacaacc actacacgca gaagagcctc tccctgtctc cgggtaaatg 3361 ataactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag 3421 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 3481 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 3541 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 3601 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 3661 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 3721 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 3781 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 3841 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 3901 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 3961 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 4021 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 4081 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 4141 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 4201 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 4261 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 4321 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 4381 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 4441 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga 4501 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 4561 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 4621 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 4681 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 4741 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 4801 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 4861 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 4921 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 4981 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 5041 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 5101 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 5161 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 5221 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 5281 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 5341 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 5401 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 5461 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 5521 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 5581 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 5641 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 5701 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 5761 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 5821 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 5881 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 5941 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 6001 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 6061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 6121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 6181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 6241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 6301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 6361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 6421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 6481 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 6541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 6601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 6661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 6721 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 6781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 6841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 6901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 6961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 7021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 7081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 7141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 7201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 7261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 7321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 7381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 7441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 7501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 7561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 7621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 7681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 7741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 7801 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat 7861 ctgctctgat gccgcatagt taagccagta t //