LOCUS       sequence-292618-        7915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgA1) high 
            affinity variant 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7915)
  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 7915)
  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-sACE2v3-Fc(IgA1)
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     654..674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     748..767
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        748..766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3391..3408)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3397..3621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3667..4095
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3754..3773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3964..3985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4104..4124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4109..4438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4289..4424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4351..4370
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4505..5299
                     /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(4559..4578)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5169..5188
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5473..5594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5510..5529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5564..5583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5643..5659)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5643..5659)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5656..5678)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5667..5683
                     /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(5691..5697,5698..5715,5716..5721))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5736..5757
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5874..5891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6045..6633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6125..6144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6804..7595,7596..7664))
                     /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     7427..7446
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7665..7769)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7844..7863)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
       61 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      121 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      661 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      721 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      781 ctagcgccac catgtcaagc tcttcctggc tccttctcag ccttgttgct gtaactgctg
      841 ctcagtccac cattgaggaa caggtcaagt actttttgga caagtttaac cacgaagccg
      901 aagacctgtt ctatcaaagt tcacttgctt cttggaatta taacaccaat attactgaag
      961 agaatgtcca aaacatgaat aatgctgggg acaaatggtc tgccttttta aaggaacagt
     1021 ccacacttgc ccaaatgtat ccactacaag aaattcagaa tctccaagtc aagcttcagc
     1081 tgcaggctct tcagcaaaat gggtcttcag tgctctcaga agacaagagc aaacggttga
     1141 acacaattct aaatacaatg agcaccatct acagtactgg aaaagtttgt aacccagata
     1201 atccacaaga atgcttatta cttgaaccag gtttgaatga aataatggca aacagtttag
     1261 actacaatga gaggctctgg gcttgggaaa gctggagatc tgaggtcggc aagcagctga
     1321 ggccattata tgaagagtat gtggtcttga aaaatgagat ggcaagagca aatcattatg
     1381 aggactatgg ggattattgg agaggagact atgaagtaaa tggggtagat ggctatgact
     1441 acagccgcgg ccagttgatt gaagatgtgg aacatacctt tgaagagatt aaaccattat
     1501 atgaacatct tcatgcctat gtgagggcaa agttgatgaa tgcctatcct tcctatatca
     1561 gtccaattgg atgcctccct gctcatttgc ttggtgatat gtggggtaga ttttggacaa
     1621 atctgtactc tttgacagtt ccctttggac agaaaccaaa catagatgtt actgatgcaa
     1681 tggtggacca ggcctgggat gcacagagaa tattcaagga ggccgagaag ttctttgtat
     1741 ctgttggtct tcctaatatg actccaggat tctgggaaaa ttccatgcta acggacccag
     1801 gaaatgttca gaaagcagtc tgccatccca cagcttggga cctggggaag ggcgacttca
     1861 ggattcttat gtgcacaaag gtgacaatgg acgacttcct gacagctcat catgagatgg
     1921 ggcatatcca gtatgatatg gcatatcttg cacaaccttt tctgctaaga aatggagcta
     1981 atgaaggatt ccatgaagct gttggggaaa tcatgtcact ttctgcagcc acacctaagc
     2041 atttaaaatc cattggtctt ctgtcacccg attttcaaga agacaatgaa acagaaataa
     2101 acttcctgct caaacaagca ctcacgattg ttgggactct gccatttact tacatgttag
     2161 agaagtggag gtggatggtc tttaaagggg aaattcccaa agaccagtgg atgaaaaagt
     2221 ggtgggagat gaagcgagag atagttgggg tggtggaacc tgtgccccat gatgaaacat
     2281 actgtgaccc cgcatctctg ttccatgttt ctaatgatta ctcattcatt cgatattaca
     2341 caaggaccct ttaccaattc cagtttcaag aagcactttg tcaagcagct aaacatgaag
     2401 gccctctgca caaatgtgac atctcaaact ctacagaagc tggacagaaa ctgttcaata
     2461 tgctgaggct tggaaaatca gaaccctgga ccctagcatt ggaaaatgtt gtaggagcaa
     2521 agaacatgaa tgtaaggcca ctgctcaact actttgagcc cttatttacc tggctgaaag
     2581 accagaacaa gaattctttt gtgggatgga gtaccgactg gagtccatat gcagacggat
     2641 ccggtggagg atcttgccat ccaagactta gtctccatcg cccagctctt gaagatttgt
     2701 tgctgggaag cgaggccaat ctcacctgca cactgactgg ccttagagac gcatccggcg
     2761 tcacgttcac atggacgcct agttcaggaa aaagcgccgt acaggggcca cctgaaaggg
     2821 acctgtgtgg gtgctattca gttagttctg tcctgcctgg atgcgcggaa ccgtggaatc
     2881 atgggaaaac ttttacctgc acggcggctt atccagaaag caaaacccct ctcacggcga
     2941 cccttagcaa gagcggaaat acttttcggc cagaagttca cttgttgcct ccgccgagcg
     3001 aggaattggc acttaatgag ctcgttaccc tcacttgcct tgcacgaggg ttctctccta
     3061 aggatgtgct cgttcgatgg ttgcagggct cacaggaact tcctagggag aaatacttga
     3121 cctgggcctc tagacaggaa ccgtctcaag gcaccacaac atttgcggtt acaagtatct
     3181 tgcgcgtagc agctgaggac tggaagaagg gtgacacctt ttcctgtatg gttggacatg
     3241 aggccctgcc acttgcattt acccagaaaa ctatagatag gcttgctggg aagcccacgc
     3301 acgtcaacgt cagtgtggta atggccgaag tggatggtac atgttactga taactcgagt
     3361 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3421 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3481 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3541 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3601 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3661 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3721 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3781 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3841 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3901 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3961 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4021 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4081 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4141 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4201 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4261 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4321 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4381 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4441 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     4501 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4561 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4621 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4681 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4741 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4801 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4861 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4921 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4981 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5041 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5101 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5161 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5221 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5281 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5341 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5401 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5461 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5521 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5581 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5641 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5701 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5761 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5821 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6721 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6781 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6841 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6901 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6961 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7021 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7081 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7141 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7201 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7261 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7321 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7381 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7441 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7501 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7561 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7621 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7681 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7741 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7801 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     7861 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagta
//
