LOCUS       sequence-292596-        7915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgA1) high 
            affinity variant 5.
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-sACE2v5-Fc(IgA1)
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3272..3289)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3278..3502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3548..3976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3635..3654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3845..3866
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3985..4005)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3990..4319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4170..4305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4232..4251
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4386..5180
                     /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(4440..4459)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5050..5069
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5354..5475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5391..5410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5445..5464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5524..5540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5524..5540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5537..5559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5548..5564
                     /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(5572..5578,5579..5596,5597..5602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5617..5638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5755..5772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5926..6514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6006..6025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6685..7476,7477..7545))
                     /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     7308..7327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7546..7650)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7725..7744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgcca ccatgtcaag ctcttcctgg ctccttctca gccttgttgc tgtaactgct
      721 gctcagtcca ccattgagga acaggtcaag acatttttgg acaagtttaa ccacgaagcc
      781 gaagacctgt tctatcaaag ttcacttgct tcttggaatt ataacaccaa tattactgaa
      841 gagaatgtcc aaaacatgaa taatgctggg gacaaatggt ctgccttttt aaaggaacag
      901 tccacacttg cccaaatgta tccactacaa gaaattcaga atctccaagt caagcttcag
      961 ctgcaggctc ttcagcaaaa tgggtcttca gtgctctcag aagacaagag caaacggttg
     1021 aacacaattc taaatacaat gagcaccatc tacagtactg gaaaagtttg taacccagat
     1081 aatccacaag aatgcttatt acttgaacca ggtttgaatg aaataatggc aaacagttta
     1141 gactacaatg agaggctctg ggcttgggaa agctggagat ctgaggtcgg caagcagctg
     1201 aggccattat atgaagagta tgtggtcttg aaaaatgaga tggcaagagc aaatcattat
     1261 gaggactatg gggattattg gagaggagac tatgaagtaa atggggtaga tggctatgac
     1321 tacagccgcg gccagttgat tgaagatgtg gaacatacct ttgaagagat taaaccatta
     1381 tatgaacatc ttcatgccta tgtgagggca aagttgatga atgcctatcc ttcctatatc
     1441 agtccaattg gatgcctccc tgctcatttg cttggtgata tgtggggtag attttggaca
     1501 aatctgtact ctttgacagt tccctttgga cagaaaccaa acatagatgt tactgatgca
     1561 atggtggacc aggcctggga tgcacagaga atattcaagg aggccgagaa gttctttgta
     1621 tctgttggtc ttcctaatat gactcaagga ttctgggaaa attccatgct aacggaccca
     1681 ggaaatgttc agaaagcagt ctgccatccc acagcttggg acctggggaa gggcgacttc
     1741 aggattctta tgtgcacaaa ggtgacaatg gacgacttcc tgacagctca tcatgagatg
     1801 gggcatatcc agtatgatat ggcatatctt gcacaacctt ttctgctaag aaatggagct
     1861 aatgaaggat tccatgaagc tgttggggaa atcatgtcac tttctgcagc cacacctaag
     1921 catttaaaat ccattggtct tctgtcaccc gattttcaag aagacaatga aacagaaata
     1981 aacttcctgc tcaaacaagc actcacgatt gttgggactc tgccatttac ttacatgtta
     2041 gagaagtgga ggtggatggt ctttaaaggg gaaattccca aagaccagtg gatgaaaaag
     2101 tggtgggaga tgaagcgaga gatagttggg gtggtggaac ctgtgcccca tgatgaaaca
     2161 tactgtgacc ccgcatctct gttccatgtt tctaatgatt actcattcat tcgatattac
     2221 acaaggaccc tttaccaatt ccagtttcaa gaagcacttt gtcaagcagc taaacatgaa
     2281 ggccctctgc acaaatgtga catctcaaac tctacagaag ctggacagaa actgttcaat
     2341 atgctgaggc ttggaaaatc agaaccctgg accctagcat tggaaaatgt tgtaggagca
     2401 aagaacatga atgtaaggcc actgctcaac tactttgagc ccttatttac ctggctgaaa
     2461 gaccagaaca agaattcttt tgtgggatgg agtaccgact ggagtccata tgcagacgga
     2521 tccggtggag gatcttgcca tccaagactt agtctccatc gcccagctct tgaagatttg
     2581 ttgctgggaa gcgaggccaa tctcacctgc acactgactg gccttagaga cgcatccggc
     2641 gtcacgttca catggacgcc tagttcagga aaaagcgccg tacaggggcc acctgaaagg
     2701 gacctgtgtg ggtgctattc agttagttct gtcctgcctg gatgcgcgga accgtggaat
     2761 catgggaaaa cttttacctg cacggcggct tatccagaaa gcaaaacccc tctcacggcg
     2821 acccttagca agagcggaaa tacttttcgg ccagaagttc acttgttgcc tccgccgagc
     2881 gaggaattgg cacttaatga gctcgttacc ctcacttgcc ttgcacgagg gttctctcct
     2941 aaggatgtgc tcgttcgatg gttgcagggc tcacaggaac ttcctaggga gaaatacttg
     3001 acctgggcct ctagacagga accgtctcaa ggcaccacaa catttgcggt tacaagtatc
     3061 ttgcgcgtag cagctgagga ctggaagaag ggtgacacct tttcctgtat ggttggacat
     3121 gaggccctgc cacttgcatt tacccagaaa actatagata ggcttgctgg gaagcccacg
     3181 cacgtcaacg tcagtgtggt aatggccgaa gtggatggta catgttactg ataactcgag
     3241 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3301 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3361 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3421 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3481 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3541 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3601 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3661 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3721 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3781 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3841 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3901 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3961 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4021 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4081 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4141 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4201 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4261 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4321 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4381 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4441 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4501 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4561 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4621 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4681 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4741 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4801 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4861 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4921 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4981 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5041 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5101 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5161 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5221 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5281 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5341 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5401 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5461 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5521 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5581 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5641 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5701 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5761 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6361 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6481 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6601 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6661 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6721 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6781 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6841 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6901 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6961 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7021 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7081 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7141 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7201 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7261 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7321 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7381 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7441 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7501 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7561 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7621 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7681 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     7741 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7801 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7861 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
