LOCUS       sequence-292590-        7210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2 (protease domain) high
            affinity variant 5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7210)
  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 7210)
  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
FEATURES             Location/Qualifiers
     source          1..7210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        172..551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        552..755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     706..726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     800..819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        800..818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2738..2755)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2744..2968
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3014..3442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3101..3120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3311..3332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3451..3471)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3456..3785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3636..3771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3698..3717
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3852..4646
                     /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(3906..3925)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4516..4535
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4820..4941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4857..4876)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4911..4930
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4990..5006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4990..5006)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5003..5025)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5014..5030
                     /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(5038..5044,5045..5062,5063..5068))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5083..5104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5221..5238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5392..5980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5472..5491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6151..6942,6943..7011))
                     /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     6774..6793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7012..7116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7191..7210)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
       61 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      121 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      541 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      601 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      661 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      721 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      781 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgcc
      841 accatgtcaa gctcttcctg gctccttctc agccttgttg ctgtaactgc tgctcagtcc
      901 accattgagg aacaggtcaa gacatttttg gacaagttta accacgaagc cgaagacctg
      961 ttctatcaaa gttcacttgc ttcttggaat tataacacca atattactga agagaatgtc
     1021 caaaacatga ataatgctgg ggacaaatgg tctgcctttt taaaggaaca gtccacactt
     1081 gcccaaatgt atccactaca agaaattcag aatctccaag tcaagcttca gctgcaggct
     1141 cttcagcaaa atgggtcttc agtgctctca gaagacaaga gcaaacggtt gaacacaatt
     1201 ctaaatacaa tgagcaccat ctacagtact ggaaaagttt gtaacccaga taatccacaa
     1261 gaatgcttat tacttgaacc aggtttgaat gaaataatgg caaacagttt agactacaat
     1321 gagaggctct gggcttggga aagctggaga tctgaggtcg gcaagcagct gaggccatta
     1381 tatgaagagt atgtggtctt gaaaaatgag atggcaagag caaatcatta tgaggactat
     1441 ggggattatt ggagaggaga ctatgaagta aatggggtag atggctatga ctacagccgc
     1501 ggccagttga ttgaagatgt ggaacatacc tttgaagaga ttaaaccatt atatgaacat
     1561 cttcatgcct atgtgagggc aaagttgatg aatgcctatc cttcctatat cagtccaatt
     1621 ggatgcctcc ctgctcattt gcttggtgat atgtggggta gattttggac aaatctgtac
     1681 tctttgacag ttccctttgg acagaaacca aacatagatg ttactgatgc aatggtggac
     1741 caggcctggg atgcacagag aatattcaag gaggccgaga agttctttgt atctgttggt
     1801 cttcctaata tgactcaagg attctgggaa aattccatgc taacggaccc aggaaatgtt
     1861 cagaaagcag tctgccatcc cacagcttgg gacctgggga agggcgactt caggattctt
     1921 atgtgcacaa aggtgacaat ggacgacttc ctgacagctc atcatgagat ggggcatatc
     1981 cagtatgata tggcatatct tgcacaacct tttctgctaa gaaatggagc taatgaagga
     2041 ttccatgaag ctgttgggga aatcatgtca ctttctgcag ccacacctaa gcatttaaaa
     2101 tccattggtc ttctgtcacc cgattttcaa gaagacaatg aaacagaaat aaacttcctg
     2161 ctcaaacaag cactcacgat tgttgggact ctgccattta cttacatgtt agagaagtgg
     2221 aggtggatgg tctttaaagg ggaaattccc aaagaccagt ggatgaaaaa gtggtgggag
     2281 atgaagcgag agatagttgg ggtggtggaa cctgtgcccc atgatgaaac atactgtgac
     2341 cccgcatctc tgttccatgt ttctaatgat tactcattca ttcgatatta cacaaggacc
     2401 ctttaccaat tccagtttca agaagcactt tgtcaagcag ctaaacatga aggccctctg
     2461 cacaaatgtg acatctcaaa ctctacagaa gctggacaga aactgttcaa tatgctgagg
     2521 cttggaaaat cagaaccctg gaccctagca ttggaaaatg ttgtaggagc aaagaacatg
     2581 aatgtaaggc cactgctcaa ctactttgag cccttattta cctggctgaa agaccagaac
     2641 aagaattctt ttgtgggatg gagtaccgac tggagtccat atgcagacgg atcctgataa
     2701 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2761 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2821 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2881 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2941 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3001 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3061 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3121 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3181 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3241 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3301 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3361 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3421 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3601 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3661 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3721 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3781 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3841 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3901 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3961 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4021 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4081 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4141 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4201 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4261 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4321 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4381 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4441 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4501 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4561 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4621 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4681 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4741 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4801 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4861 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4921 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4981 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5041 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5101 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5161 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5221 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5281 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5581 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5821 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5941 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6001 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6061 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6121 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6181 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6241 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6301 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6361 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6421 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6481 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6541 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6601 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6661 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6721 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6781 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6841 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6901 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6961 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7021 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7141 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     7201 tctgatgccg
//
