LOCUS       sequence-292592-        7915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgA1).
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-sACE2(WT)-Fc(IgA1)
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        449..652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     603..623
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     697..716
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        697..715
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3340..3357)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3346..3570
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3616..4044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3703..3722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3913..3934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4053..4073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4058..4387
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4238..4373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4300..4319
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4454..5248
                     /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(4508..4527)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5118..5137
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5422..5543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5459..5478)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5513..5532
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5592..5608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5592..5608)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5605..5627)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5616..5632
                     /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(5640..5646,5647..5664,5665..5670))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5685..5706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5823..5840)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5994..6582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6074..6093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6753..7544,7545..7613))
                     /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     7376..7395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7614..7718)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7793..7812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
       61 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      121 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      181 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      301 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      361 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      421 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      481 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      541 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      601 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
      661 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      721 ccaagctggc tagcgccacc atgtcaagct cttcctggct ccttctcagc cttgttgctg
      781 taactgctgc tcagtccacc attgaggaac aggccaagac atttttggac aagtttaacc
      841 acgaagccga agacctgttc tatcaaagtt cacttgcttc ttggaattat aacaccaata
      901 ttactgaaga gaatgtccaa aacatgaata atgctgggga caaatggtct gcctttttaa
      961 aggaacagtc cacacttgcc caaatgtatc cactacaaga aattcagaat ctcacagtca
     1021 agcttcagct gcaggctctt cagcaaaatg ggtcttcagt gctctcagaa gacaagagca
     1081 aacggttgaa cacaattcta aatacaatga gcaccatcta cagtactgga aaagtttgta
     1141 acccagataa tccacaagaa tgcttattac ttgaaccagg tttgaatgaa ataatggcaa
     1201 acagtttaga ctacaatgag aggctctggg cttgggaaag ctggagatct gaggtcggca
     1261 agcagctgag gccattatat gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa
     1321 atcattatga ggactatggg gattattgga gaggagacta tgaagtaaat ggggtagatg
     1381 gctatgacta cagccgcggc cagttgattg aagatgtgga acataccttt gaagagatta
     1441 aaccattata tgaacatctt catgcctatg tgagggcaaa gttgatgaat gcctatcctt
     1501 cctatatcag tccaattgga tgcctccctg ctcatttgct tggtgatatg tggggtagat
     1561 tttggacaaa tctgtactct ttgacagttc cctttggaca gaaaccaaac atagatgtta
     1621 ctgatgcaat ggtggaccag gcctgggatg cacagagaat attcaaggag gccgagaagt
     1681 tctttgtatc tgttggtctt cctaatatga ctcaaggatt ctgggaaaat tccatgctaa
     1741 cggacccagg aaatgttcag aaagcagtct gccatcccac agcttgggac ctggggaagg
     1801 gcgacttcag gattcttatg tgcacaaagg tgacaatgga cgacttcctg acagctcatc
     1861 atgagatggg gcatatccag tatgatatgg catatgctgc acaacctttt ctgctaagaa
     1921 atggagctaa tgaaggattc catgaagctg ttggggaaat catgtcactt tctgcagcca
     1981 cacctaagca tttaaaatcc attggtcttc tgtcacccga ttttcaagaa gacaatgaaa
     2041 cagaaataaa cttcctgctc aaacaagcac tcacgattgt tgggactctg ccatttactt
     2101 acatgttaga gaagtggagg tggatggtct ttaaagggga aattcccaaa gaccagtgga
     2161 tgaaaaagtg gtgggagatg aagcgagaga tagttggggt ggtggaacct gtgccccatg
     2221 atgaaacata ctgtgacccc gcatctctgt tccatgtttc taatgattac tcattcattc
     2281 gatattacac aaggaccctt taccaattcc agtttcaaga agcactttgt caagcagcta
     2341 aacatgaagg ccctctgcac aaatgtgaca tctcaaactc tacagaagct ggacagaaac
     2401 tgttcaatat gctgaggctt ggaaaatcag aaccctggac cctagcattg gaaaatgttg
     2461 taggagcaaa gaacatgaat gtaaggccac tgctcaacta ctttgagccc ttatttacct
     2521 ggctgaaaga ccagaacaag aattcttttg tgggatggag taccgactgg agtccatatg
     2581 cagacggatc cggtggagga tcttgccatc caagacttag tctccatcgc ccagctcttg
     2641 aagatttgtt gctgggaagc gaggccaatc tcacctgcac actgactggc cttagagacg
     2701 catccggcgt cacgttcaca tggacgccta gttcaggaaa aagcgccgta caggggccac
     2761 ctgaaaggga cctgtgtggg tgctattcag ttagttctgt cctgcctgga tgcgcggaac
     2821 cgtggaatca tgggaaaact tttacctgca cggcggctta tccagaaagc aaaacccctc
     2881 tcacggcgac ccttagcaag agcggaaata cttttcggcc agaagttcac ttgttgcctc
     2941 cgccgagcga ggaattggca cttaatgagc tcgttaccct cacttgcctt gcacgagggt
     3001 tctctcctaa ggatgtgctc gttcgatggt tgcagggctc acaggaactt cctagggaga
     3061 aatacttgac ctgggcctct agacaggaac cgtctcaagg caccacaaca tttgcggtta
     3121 caagtatctt gcgcgtagca gctgaggact ggaagaaggg tgacaccttt tcctgtatgg
     3181 ttggacatga ggccctgcca cttgcattta cccagaaaac tatagatagg cttgctggga
     3241 agcccacgca cgtcaacgtc agtgtggtaa tggccgaagt ggatggtaca tgttactgat
     3301 aactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3361 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3421 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3481 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3541 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3601 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3661 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3721 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3781 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3841 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3901 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3961 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4021 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4081 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4141 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4201 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4261 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4321 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4381 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4441 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4501 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4561 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4621 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4681 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4741 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4801 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4861 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4921 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4981 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5041 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5101 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5161 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5221 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5281 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5341 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5401 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5461 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5521 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5581 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5641 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5701 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5761 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5821 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5881 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5941 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6001 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6061 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6121 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6181 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6241 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6301 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6361 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6421 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6481 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6541 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6601 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6661 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6721 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6781 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6841 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6901 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6961 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7021 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7081 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7141 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7201 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7261 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7321 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7381 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7441 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7501 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7561 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7621 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7681 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7741 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     7801 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7861 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattg
//
