LOCUS       sequence-292595-        7915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgA1) high 
            affinity variant 1.
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-sACE2v1-Fc(IgA1)
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     781..800
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        781..799
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3424..3441)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3430..3654
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3700..4128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3787..3806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3997..4018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4137..4157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4142..4471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4322..4457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4384..4403
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4538..5332
                     /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(4592..4611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5202..5221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5506..5627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5543..5562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5597..5616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5676..5692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5676..5692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5689..5711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5700..5716
                     /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(5724..5730,5731..5748,5749..5754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5769..5790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5907..5924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6078..6666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6158..6177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6837..7628,7629..7697))
                     /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     7460..7479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7698..7802)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7877..7896)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
       61 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
      121 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
      781 taatacgact cactataggg agacccaagc tggctagcgc caccatgtca agctcttcct
      841 ggctccttct cagccttgtt gctgtaactg ctgctcagtc caccattgag gaacaggcca
      901 agacattttt ggacaagttt aacgccgaag ccgaagacct gttctatcaa agttcacttg
      961 cttcttggaa ttataacacc aatattactg aagagaatgt ccaaaacatg aataatgctg
     1021 gggacaaatg gtctgccttt ttaaaggaac agtccacact tgcccaaatg tatccactac
     1081 aagaaattca gaatctccaa gtcaagcttc agctgcaggc tcttcagcaa aatgggtctt
     1141 cagtgctctc agaagacaag agcaaacggt tgaacacaat tctaaataca atgagcacca
     1201 tctacagtac tggaaaagtt tgtaacccag ataatccaca agaatgctta ttacttgaac
     1261 caggtttgaa tgaaataatg gcaaacagtt tagactacaa tgagaggctc tgggcttggg
     1321 aaagctggag atctgaggtc ggcaagcagc tgaggccatt atatgaagag tatgtggtct
     1381 tgaaaaatga gatggcaaga gcaaatcatt atgaggacta tggggattat tggagaggag
     1441 actatgaagt aaatggggta gatggctatg actacagccg cggccagttg attgaagatg
     1501 tggaacatac ctttgaagag attaaaccat tatatgaaca tcttcatgcc tatgtgaggg
     1561 caaagttgat gaatgcctat ccttcctata tcagtccaat tggatgcctc cctgctcatt
     1621 tgcttggtga tatgtggggt agattttgga caaatctgta ctctttgaca gttccctttg
     1681 gacagaaacc aaacatagat gttactgatg caatggtgga ccaggcctgg gatgcacaga
     1741 gaatattcaa ggaggccgag aagttctttg tatctgttgg tcttcctaat atgactccag
     1801 gattctggga aaattccatg ctaacggacc caggaaatgt tcagaaagca gtctgccatc
     1861 ccacagcttg ggacctgggg aagggcgact tcaggattct tatgtgcaca aaggtgacaa
     1921 tggacgactt cctgacagct catcatgaga tggggcatat ccagtatgat atggcatatc
     1981 ttgcacaacc ttttctgcta agaaatggag ctaatgaagg attccatgaa gctgttgggg
     2041 aaatcatgtc actttctgca gccacaccta agcatttaaa atccattggt cttctgtcac
     2101 ccgattttca agaagacaat gaaacagaaa taaacttcct gctcaaacaa gcactcacga
     2161 ttgttgggac tctgccattt acttacatgt tagagaagtg gaggtggatg gtctttaaag
     2221 gggaaattcc caaagaccag tggatgaaaa agtggtggga gatgaagcga gagatagttg
     2281 gggtggtgga acctgtgccc catgatgaaa catactgtga ccccgcatct ctgttccatg
     2341 tttctaatga ttactcattc attcgatatt acacaaggac cctttaccaa ttccagtttc
     2401 aagaagcact ttgtcaagca gctaaacatg aaggccctct gcacaaatgt gacatctcaa
     2461 actctacaga agctggacag aaactgttca atatgctgag gcttggaaaa tcagaaccct
     2521 ggaccctagc attggaaaat gttgtaggag caaagaacat gaatgtaagg ccactgctca
     2581 actactttga gcccttattt acctggctga aagaccagaa caagaattct tttgtgggat
     2641 ggagtaccga ctggagtcca tatgcagacg gatccggtgg aggatcttgc catccaagac
     2701 ttagtctcca tcgcccagct cttgaagatt tgttgctggg aagcgaggcc aatctcacct
     2761 gcacactgac tggccttaga gacgcatccg gcgtcacgtt cacatggacg cctagttcag
     2821 gaaaaagcgc cgtacagggg ccacctgaaa gggacctgtg tgggtgctat tcagttagtt
     2881 ctgtcctgcc tggatgcgcg gaaccgtgga atcatgggaa aacttttacc tgcacggcgg
     2941 cttatccaga aagcaaaacc cctctcacgg cgacccttag caagagcgga aatacttttc
     3001 ggccagaagt tcacttgttg cctccgccga gcgaggaatt ggcacttaat gagctcgtta
     3061 ccctcacttg ccttgcacga gggttctctc ctaaggatgt gctcgttcga tggttgcagg
     3121 gctcacagga acttcctagg gagaaatact tgacctgggc ctctagacag gaaccgtctc
     3181 aaggcaccac aacatttgcg gttacaagta tcttgcgcgt agcagctgag gactggaaga
     3241 agggtgacac cttttcctgt atggttggac atgaggccct gccacttgca tttacccaga
     3301 aaactataga taggcttgct gggaagccca cgcacgtcaa cgtcagtgtg gtaatggccg
     3361 aagtggatgg tacatgttac tgataactcg agtctagagg gcccgtttaa acccgctgat
     3421 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3481 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3541 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3601 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3661 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3721 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3781 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3841 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3901 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3961 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4021 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4081 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4141 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4201 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4321 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4381 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4441 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4501 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4561 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4621 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4681 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4741 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4801 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4861 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4921 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4981 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5041 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5101 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5161 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5221 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5281 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5341 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5401 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5461 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5521 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5581 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5641 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5701 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5761 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5821 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5881 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5941 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6001 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6061 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6121 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6181 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6241 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6301 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6361 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6421 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6481 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6541 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6601 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6661 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6721 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6781 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6841 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6901 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6961 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7021 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7081 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7141 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7201 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7261 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7321 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7381 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7441 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7501 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7561 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7621 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7681 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7741 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7801 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7861 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatct
//
