LOCUS Exported 7813 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express human ACE2 with C9 tag at C-terminal in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA3.1-hACE2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7813) AUTHORS Shang J, Ye G, Shi K, Wan Y, Luo C, Aihara H, Geng Q, Auerbach A, Li F TITLE Structural basis of receptor recognition by SARS-CoV-2. JOURNAL Nature. 2020 Mar 30. pii: 10.1038/s41586-020-2179-y. doi: 10.1038/s41586-020-2179-y. PUBMED 32225175 REFERENCE 2 (bases 1 to 7813) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7813 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 170..549 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 550..753 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 704..724 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 798..817 /label=T7 /note="T7 promoter, forward primer" promoter 798..816 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3341..3358) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3347..3571 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3617..4045 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3704..3723) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3914..3935 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4054..4074) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4059..4388 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4239..4374 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4301..4320 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4509..4528) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5117..5136 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5421..5542 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5458..5477) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5512..5531 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5591..5607) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5591..5607) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5604..5626) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5615..5631 /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(5639..5669) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5684..5705 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5822..5839) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5993..6581) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6073..6092) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6752..7612) /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 7375..7394 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7613..7717) /gene="bla" /label=AmpR promoter primer_bind complement(7792..7811) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc 61 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg 121 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 181 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 241 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 301 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 361 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 421 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 481 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 541 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 601 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 661 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 721 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct 781 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 841 aacgggccct ctagaccatg tcaagctctt cctggctcct tctcagcctt gttgctgtaa 901 ctgctgctca gtccaccatt gaggaacagg ccaagacatt tttggacaag tttaaccacg 961 aagccgaaga cctgttctat caaagttcac ttgcttcttg gaattataac accaatatta 1021 ctgaagagaa tgtccaaaac atgaataatg ctggggacaa atggtctgcc tttttaaagg 1081 aacagtccac acttgcccaa atgtatccac tacaagaaat tcagaatctc acagtcaagc 1141 ttcagctgca ggctcttcag caaaatgggt cttcagtgct ctcagaagac aagagcaaac 1201 ggttgaacac aattctaaat acaatgagca ccatctacag tactggaaaa gtttgtaacc 1261 cagataatcc acaagaatgc ttattacttg aaccaggttt gaatgaaata atggcaaaca 1321 gtttagacta caatgagagg ctctgggctt gggaaagctg gagatctgag gtcggcaagc 1381 agctgaggcc attatatgaa gagtatgtgg tcttgaaaaa tgagatggca agagcaaatc 1441 attatgagga ctatggggat tattggagag gagactatga agtaaatggg gtagatggct 1501 atgactacag ccgcggccag ttgattgaag atgtggaaca tacctttgaa gagattaaac 1561 cattatatga acatcttcat gcctatgtga gggcaaagtt gatgaatgcc tatccttcct 1621 atatcagtcc aattggatgc ctccctgctc atttgcttgg tgatatgtgg ggtagatttt 1681 ggacaaatct gtactctttg acagttccct ttggacagaa accaaacata gatgttactg 1741 atgcaatggt ggaccaggcc tgggatgcac agagaatatt caaggaggcc gagaagttct 1801 ttgtatctgt tggtcttcct aatatgactc aaggattctg ggaaaattcc atgctaacgg 1861 acccaggaaa tgttcagaaa gcagtctgcc atcccacagc ttgggacctg gggaagggcg 1921 acttcaggat ccttatgtgc acaaaggtga caatggacga cttcctgaca gctcatcatg 1981 agatggggca tatccagtat gatatggcat atgctgcaca accttttctg ctaagaaatg 2041 gagctaatga aggattccat gaagctgttg gggaaatcat gtcactttct gcagccacac 2101 ctaagcattt aaaatccatt ggtcttctgt cacccgattt tcaagaagac aatgaaacag 2161 aaataaactt cctgctcaaa caagcactca cgattgttgg gactctgcca tttacttaca 2221 tgttagagaa gtggaggtgg atggtcttta aaggggaaat tcccaaagac cagtggatga 2281 aaaagtggtg ggagatgaag cgagagatag ttggggtggt ggaacctgtg ccccatgatg 2341 aaacatactg tgaccccgca tctctgttcc atgtttctaa tgattactca ttcattcgat 2401 attacacaag gaccctttac caattccagt ttcaagaagc actttgtcaa gcagctaaac 2461 atgaaggccc tctgcacaaa tgtgacatct caaactctac agaagctgga cagaaactgt 2521 tcaatatgct gaggcttgga aaatcagaac cctggaccct agcattggaa aatgttgtag 2581 gagcaaagaa catgaatgta aggccactgc tcaactactt tgagccctta tttacctggc 2641 tgaaagacca gaacaagaat tcttttgtgg gatggagtac cgactggagt ccatatgcag 2701 accaaagcat caaagtgagg ataagcctaa aatcagctct tggagataaa gcatatgaat 2761 ggaacgacaa tgaaatgtac ctgttccgat catctgttgc atatgctatg aggcagtact 2821 ttttaaaagt aaaaaatcag atgattcttt ttggggagga ggatgtgcga gtggctaatt 2881 tgaaaccaag aatctccttt aatttctttg tcactgcacc taaaaatgtg tctgatatca 2941 ttcctagaac tgaagttgaa aaggccatca ggatgtcccg gagccgtatc aatgatgctt 3001 tccgtctgaa tgacaacagc ctagagtttc tggggataca gccaacactt ggacctccta 3061 accagccccc tgtttccata tggctgattg tttttggagt tgtgatggga gtgatagtgg 3121 ttggcattgt catcctgatc ttcactggga tcagagatcg gaagaagaaa aataaagcaa 3181 gaagtggaga aaatccttat gcctccatcg atattagcaa aggagaaaat aatccaggat 3241 tccaaaacac tgatgatgtt cagacctcct ttggtaccga gacctcccag gtggcgcccg 3301 cttaagcttg gatccgcgtt aagtttaaac cgctgatcag cctcgactgt gccttctagt 3361 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 3421 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 3481 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 3541 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 3601 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3661 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3721 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 3781 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3841 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 3901 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3961 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 4021 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 4081 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 4141 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 4201 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 4261 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4321 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 4381 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 4441 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 4501 tggagaggct attcggctat gactgggcaa cagacaatcg gctgctctga tgccgccgtg 4561 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 4621 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 4681 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 4741 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 4801 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 4861 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 4921 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 4981 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 5041 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 5101 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 5161 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 5221 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 5281 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 5341 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 5401 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 5461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 5521 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 5581 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 5641 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 5701 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 5761 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 5821 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 5881 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 6181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 6421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 6601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 6661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 6721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 6781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 6841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 6901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 6961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 7021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 7081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 7141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 7201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 7261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 7321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 7381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 7441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 7501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 7561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 7621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 7681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 7741 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg 7801 ctctgatgcc gca //