LOCUS       sequence-301647-        7558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for dimeric soluble ACE2 high affinity 
            variant 2.4 (untagged).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7558)
  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 7558)
  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-sACE2v2.4(732)
FEATURES             Location/Qualifiers
     source          1..7558
                     /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(2915..2932)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2921..3145
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3191..3619
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3278..3297)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3488..3509
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3628..3648)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3633..3962
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3813..3948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3875..3894
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4029..4823
                     /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(4083..4102)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4693..4712
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4997..5118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5034..5053)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5088..5107
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5167..5183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5167..5183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5180..5202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5191..5207
                     /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(5215..5221,5222..5239,5240..5245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5260..5281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5398..5415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5569..6157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5649..5668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6328..7119,7120..7188))
                     /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     6951..6970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7189..7293)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7368..7387)
                     /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 acaggccaag tactttttgg acaagtttaa ccacgaagcc
      781 gaagacctgt tctatcaaag ttcacttgct tcttggaatt ataacaccaa tattactgaa
      841 gagaatgtcc aaaacatgaa taatgctggg gacaaatggt ctgccttttt aaaggaacag
      901 tccacaactg cccaaatgta tccactacaa gaaattcaga atctcacagt 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 ttctgggaat attccatgct aacggaccca
     1681 ggaaatgttc agaaagcagt ctgccatccc acagcttggg acctggggaa gggcgacttc
     1741 aggattctta tgtgcacaaa ggtgacaatg gacgacttcc tgacagctca tcatgagatg
     1801 gggcatatcc agtatgatat ggcatatgct 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 tgcagaccaa
     2521 agcatcaaag tgaggataag cctaaaatca gctcttggag ataaagcata tgaatggaac
     2581 gacaatgaaa tgtacctgtt ccgatcatct gttgcatatg ctatgaggca gtacttttta
     2641 aaagtaaaaa atcagatgat tctttttggg gaggaggatg tgcgagtggc taatttgaaa
     2701 ccaagaatct cctttaattt ctttgtcact gcacctaaaa atgtgtctga tatcattcct
     2761 agaactgaag ttgaaaaggc catcaggatg tcccggagcc gtatcaatga tgctttccgt
     2821 ctgaatgaca acagcctaga gtttctgggg atacagccaa cacttggatc ctgataactc
     2881 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2941 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3001 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3061 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3121 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3181 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3241 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3301 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3361 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3421 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3481 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3541 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3601 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3661 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3721 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3781 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3841 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3901 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3961 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4021 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4081 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4141 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4201 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4261 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4321 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4381 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4441 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4501 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4561 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4621 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4681 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4741 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4801 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4861 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4921 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4981 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5041 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5101 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5161 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5221 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5281 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5341 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5401 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5461 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5521 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5581 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5641 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5701 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5761 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5821 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5881 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5941 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6001 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6061 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6121 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6181 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6241 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7321 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     7381 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7441 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7501 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
