LOCUS       sequence-292599-        7891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgG1) high 
            affinity 6.
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-sACE2v6-Fc(IgG1)
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2868..2885)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2874..3098
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3144..3572
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3231..3250)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3441..3462
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3581..3601)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3586..3915
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3766..3901
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3828..3847
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3982..4776
                     /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(4036..4055)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4646..4665
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4950..5071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4987..5006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5041..5060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5120..5136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5120..5136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5133..5155)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5144..5160
                     /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(5168..5174,5175..5192,5193..5198))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5213..5234
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5351..5368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5522..6110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5602..5621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6281..7072,7073..7141))
                     /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     6904..6923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7142..7246)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7321..7340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7512..7891
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgcca ccatgtcaag
      301 ctcttcctgg ctccttctca gccttgttgc tgtaactgct gctcagtcca ccattgagga
      361 acaggccaag tactttttgg acaagtttaa ccacgaagcc gaagacctgt tctatctgag
      421 ttcacttgct tcttggaatt ataacaccaa tattactgaa gagaatgtcc aaaacatgaa
      481 taatgctggg gacaaatggt ctgccttttt aaaggaacag tccacaactg cccaaatgta
      541 tccactacaa gaaattcaga atctccaagt caagcttcag ctgcaggctc ttcagcaaaa
      601 tgggtcttca gtgctctcag aagacaagag caaacggttg aacacaattc taaatacaat
      661 gagcaccatc tacagtactg gaaaagtttg taacccagat aatccacaag aatgcttatt
      721 acttgaacca ggtttgaatg aaataatggc aaacagttta gactacaatg agaggctctg
      781 ggcttgggaa agctggagat ctgaggtcgg caagcagctg aggccattat atgaagagta
      841 tgtggtcttg aaaaatgaga tggcaagagc aaatcattat gaggactatg gggattattg
      901 gagaggagac tatgaagtaa atggggtaga tggctatgac tacagccgcg gccagttgat
      961 tgaagatgtg gaacatacct ttgaagagat taaaccatta tatgaacatc ttcatgccta
     1021 tgtgagggca aagttgatga atgcctatcc ttcctatatc agtccaattg gatgcctccc
     1081 tgctcatttg cttggtgata tgtggggtag attttggaca aatctgtact ctttgacagt
     1141 tccctttgga cagaaaccaa acatagatgt tactgatgca atggtggacc aggcctggga
     1201 tgcacagaga atattcaagg aggccgagaa gttctttgta tctgttggtc ttcctaatat
     1261 gactccagga ttctgggaat attccatgct aacggaccca ggaaatgttc agaaagcagt
     1321 ctgccatccc acagcttggg acctggggaa gggcgacttc aggattctta tgtgcacaaa
     1381 ggtgacaatg gacgacttcc tgacagctca tcatgagatg gggcatatcc agtatgatat
     1441 ggcatatctt gcacaacctt ttctgctaag aaatggagct aatgaaggat tccatgaagc
     1501 tgttggggaa atcatgtcac tttctgcagc cacacctaag catttaaaat ccattggtct
     1561 tctgtcaccc gattttcaag aagacaatga aacagaaata aacttcctgc tcaaacaagc
     1621 actcacgatt gttgggactc tgccatttac ttacatgtta gagaagtgga ggtggatggt
     1681 ctttaaaggg gaaattccca aagaccagtg gatgaaaaag tggtgggaga tgaagcgaga
     1741 gatagttggg gtggtggaac ctgtgcccca tgatgaaaca tactgtgacc ccgcatctct
     1801 gttccatgtt tctaatgatt actcattcat tcgatattac acaaggaccc tttaccaatt
     1861 ccagtttcaa gaagcacttt gtcaagcagc taaacatgaa ggccctctgc acaaatgtga
     1921 catctcaaac tctacagaag ctggacagaa actgttcaat atgctgaggc ttggaaaatc
     1981 agaaccctgg accctagcat tggaaaatgt tgtaggagca aagaacatga atgtaaggcc
     2041 actgctcaac tactttgagc ccttatttac ctggctgaaa gaccagaaca agaattcttt
     2101 tgtgggatgg agtaccgact ggagtccata tgcagacgga tccgacaaaa ctcacacatg
     2161 cccaccgtgc ccagcacctg aactcctggg gggaccgtca gtcttcctct tccccccaaa
     2221 acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg tggtggacgt
     2281 gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg aggtgcataa
     2341 tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg tcagcgtcct
     2401 caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg tctccaacaa
     2461 agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc cccgagaacc
     2521 acaggtgtac accctgcccc catcccggga ggagatgacc aagaaccagg tcagcctgac
     2581 ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga gcaatgggca
     2641 gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct ccttcttcct
     2701 ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct tctcatgctc
     2761 cgtgatgcac gaggctctgc acaaccacta cacgcagaag agcctctccc tgtctccggg
     2821 taaatgataa ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     2881 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2941 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3001 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3061 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3121 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3181 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3241 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3301 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3361 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3421 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3481 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3541 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3661 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3721 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3781 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3841 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3901 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3961 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4021 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4081 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4141 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4201 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4261 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4321 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4381 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4441 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4501 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4561 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4621 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4681 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4741 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4801 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4861 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4921 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4981 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5041 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5101 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5161 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5221 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5281 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5341 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5401 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5461 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5521 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5581 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5641 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5701 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5761 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5821 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5881 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5941 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6001 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6061 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6121 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6181 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6241 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6301 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6361 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6421 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6481 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6541 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6601 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6661 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6721 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6781 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6841 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6901 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6961 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7021 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7081 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7141 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7201 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7261 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     7321 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     7381 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     7441 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     7501 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     7561 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     7621 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     7681 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     7741 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     7801 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     7861 catctacgta ttagtcatcg ctattaccat g
//
