LOCUS       sequence-292630-        7891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgG1) 
            glycosylation mutant T92Q.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.abc0870"; journalName: "Science"; date: "2020-09-4-
            4"; volume: "369"; issue: "6508"; pages: "1261-1265"
COMMENT     Sequence Label: pcDNA3-sACE2-T92Q-Fc(IgG1)
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        137..516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        517..720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     671..691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     765..784
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        765..783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3384..3401)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3390..3614
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3660..4088
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3747..3766)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3957..3978
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4097..4117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4102..4431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4282..4417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4344..4363
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4498..5292
                     /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(4552..4571)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5162..5181
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5466..5587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5503..5522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5557..5576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5636..5652)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5636..5652)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5649..5671)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5660..5676
                     /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(join(5684..5690,5691..5708,5709..5714))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5729..5750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5867..5884)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6038..6626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6118..6137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6797..7588,7589..7657))
                     /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     7420..7439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7658..7762)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7837..7856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
       61 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      121 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      721 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      781 agggagaccc aagctggcta gcgccaccat gtcaagctct tcctggctcc ttctcagcct
      841 tgttgctgta actgctgctc agtccaccat tgaggaacag gccaagacat ttttggacaa
      901 gtttaaccac gaagccgaag acctgttcta tcaaagttca cttgcttctt ggaattataa
      961 caccaatatt actgaagaga atgtccaaaa catgaataat gctggggaca aatggtctgc
     1021 ctttttaaag gaacagtcca cacttgccca aatgtatcca ctacaagaaa ttcagaatct
     1081 ccaagtcaag cttcagctgc aggctcttca gcaaaatggg tcttcagtgc tctcagaaga
     1141 caagagcaaa cggttgaaca caattctaaa tacaatgagc accatctaca gtactggaaa
     1201 agtttgtaac ccagataatc cacaagaatg cttattactt gaaccaggtt tgaatgaaat
     1261 aatggcaaac agtttagact acaatgagag gctctgggct tgggaaagct ggagatctga
     1321 ggtcggcaag cagctgaggc cattatatga agagtatgtg gtcttgaaaa atgagatggc
     1381 aagagcaaat cattatgagg actatgggga ttattggaga ggagactatg aagtaaatgg
     1441 ggtagatggc tatgactaca gccgcggcca gttgattgaa gatgtggaac atacctttga
     1501 agagattaaa ccattatatg aacatcttca tgcctatgtg agggcaaagt tgatgaatgc
     1561 ctatccttcc tatatcagtc caattggatg cctccctgct catttgcttg gtgatatgtg
     1621 gggtagattt tggacaaatc tgtactcttt gacagttccc tttggacaga aaccaaacat
     1681 agatgttact gatgcaatgg tggaccaggc ctgggatgca cagagaatat tcaaggaggc
     1741 cgagaagttc tttgtatctg ttggtcttcc taatatgact caaggattct gggaaaattc
     1801 catgctaacg gacccaggaa atgttcagaa agcagtctgc catcccacag cttgggacct
     1861 ggggaagggc gacttcagga ttcttatgtg cacaaaggtg acaatggacg acttcctgac
     1921 agctcatcat gagatggggc atatccagta tgatatggca tatgctgcac aaccttttct
     1981 gctaagaaat ggagctaatg aaggattcca tgaagctgtt ggggaaatca tgtcactttc
     2041 tgcagccaca cctaagcatt taaaatccat tggtcttctg tcacccgatt ttcaagaaga
     2101 caatgaaaca gaaataaact tcctgctcaa acaagcactc acgattgttg ggactctgcc
     2161 atttacttac atgttagaga agtggaggtg gatggtcttt aaaggggaaa ttcccaaaga
     2221 ccagtggatg aaaaagtggt gggagatgaa gcgagagata gttggggtgg tggaacctgt
     2281 gccccatgat gaaacatact gtgaccccgc atctctgttc catgtttcta atgattactc
     2341 attcattcga tattacacaa ggacccttta ccaattccag tttcaagaag cactttgtca
     2401 agcagctaaa catgaaggcc ctctgcacaa atgtgacatc tcaaactcta cagaagctgg
     2461 acagaaactg ttcaatatgc tgaggcttgg aaaatcagaa ccctggaccc tagcattgga
     2521 aaatgttgta ggagcaaaga acatgaatgt aaggccactg ctcaactact ttgagccctt
     2581 atttacctgg ctgaaagacc agaacaagaa ttcttttgtg ggatggagta ccgactggag
     2641 tccatatgca gacggatccg acaaaactca cacatgccca ccgtgcccag cacctgaact
     2701 cctgggggga ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc
     2761 ccggacccct gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa
     2821 gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga
     2881 gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct
     2941 gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa
     3001 aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc
     3061 ccgggaggag atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc
     3121 cagcgacatc gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac
     3181 gcctcccgtg ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa
     3241 gagcaggtgg cagcagggga acgtcttctc atgctccgtg atgcacgagg ctctgcacaa
     3301 ccactacacg cagaagagcc tctccctgtc tccgggtaaa tgataactcg agtctagagg
     3361 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3421 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3481 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3541 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3601 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3661 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3721 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3781 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3841 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3901 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3961 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4021 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4081 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4261 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4321 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4381 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4441 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4501 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4561 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4621 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4681 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4741 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4801 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4861 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4921 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4981 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5041 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5101 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5161 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5221 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5281 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5341 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5401 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5461 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5521 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5581 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5641 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5701 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5761 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5821 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5881 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5941 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6001 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6061 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6421 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6481 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6541 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6721 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7381 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7801 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     7861 gttaagccag tatctgctcc ctgcttgtgt g
//
