LOCUS       sequence-314501-        8461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for mutated Fc-fused ACE2(740).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8461)
  AUTHORS   Glasgow A, Glasgow J, Limonta D, Solomon P, Lui I, Zhang Y, Nix MA, 
            Rettko NJ, Zha S, Yamin R, Kao K, Rosenberg OS, Ravetch JV, Wiita 
            AP, Leung KK, Lim SA, Zhou XX, Hobman TC, Kortemme T, Wells JA
  TITLE     Engineered ACE2 receptor traps potently neutralize SARS-CoV-2.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Nov 10;117(45):28046-28055. doi: 
            10.1073/pnas.2016093117. Epub 2020 Oct 22.
  PUBMED    33093202
REFERENCE   2  (bases 1 to 8461)
  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: 
            "10.1073/pnas.2016093117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-11-10- 10"; volume: "117"; issue: "45"; pages: 
            "28046-28055"
COMMENT     Sequence Label: pCVD293
FEATURES             Location/Qualifiers
     source          1..8461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        54..265
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             278..546
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2849..2869
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3605..3649
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3685..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3696..3715)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3750..3769
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3910..4304
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4318..4692)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     enhancer        5285..5588
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5668..6256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5748..5767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6364..6412
                     /note="synthetic polyadenylation signal"
     primer_bind     6463..6482
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        6533..6836
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6837..7040
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6991..7011
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7085..7104
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7085..7103
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7136..8101
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             7136..8098
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             8121..8150
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             8166..8183
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8206..8223)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8212..8436
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 tgccagaaca tttctctatc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
       61 gcgcacatcg ccaacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
      121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
      181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
      241 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
      301 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
      361 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
      421 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
      481 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
      541 cagatccaag ctgtgaccgg cgcctacctg agatcaccgg cgaaggaggg ccaccatgcg
      601 aatgcagctg ctgctgctga ttgcgctgag cctggcgctg gtgaccaaca gcactagtca
      661 atctaccatc gaagagcagg ccaaaacatt cctcgacttc tttaatatcc aggctgaaga
      721 ccttttctac caatcaagtc tggctagctg gaattacaat acaaacatta cagaggagaa
      781 cgtacaaaac atgaataacg caggggacaa gtggagcgca ttccttaagg aacaaagtac
      841 ccttgcgcaa atgtatccgc tgcaagagat tcaaaacctg acggttaagc tgcaacttca
      901 ggccctccaa caaaatggaa gttcagtctt gtcagaagac aaaagcaagc gactgaacac
      961 catccttaac accatgtcaa ccatatattc aacaggtaaa gtttgcaatc cggataaccc
     1021 ccaagaatgt ttgcttcttg aacccggtct caacgaaatt atggccaaca gtcttgatta
     1081 caacgagcga ttgtgggcat gggaaagttg gaggagtgag gtaggcaaac agttgagacc
     1141 tctttatgaa gagtacgttg tccttaaaaa tgaaatggct cgcgcgaatc attatgaaga
     1201 ctatggtgac tactggaggg gggattatga ggtgaacggg gtggacggat acgattactc
     1261 taggggccag ctgatagagg atgtcgagca cacctttgag gagattaagc cgttgtacga
     1321 acatttgcac gcctatgtca gggctaagct catgaacgct tatccgagtt atatctcccc
     1381 gataggatgc ttgcctgctc acttgttggg cgatatgtgg ggacgctttt ggaccaactt
     1441 gtattccctt acggtaccgt tcggccagaa accaaatatc gacgtgacag acgcaatggt
     1501 ggatcaagca tgggatgcgc aacgaatctt caaggaggca gaaaaatttt tcgtttcagt
     1561 tggactccca aacatgacgc agggtttctg ggagaactca atgttgacag atccaggtaa
     1621 tgtgcagaaa gcggtttgcc accctactgc atgggatctt ggtaaagggg acttccgcat
     1681 actcatgtgt acgaaagtaa ctatggacga ctttcttact gcgcaccacg agatggggca
     1741 catacaatac gatatggcgt acgcagctca acctttcctt ctgcggaacg gggcgaatga
     1801 aggatttcac gaggcagtgg gtgagattat gtccctgtca gctgccactc cgaaacatct
     1861 gaaaagcatc ggcctgttga gcccagactt ccaagaagat aatgagaccg aaataaactt
     1921 ccttctgaag caagcactga ctattgtagg taccttgccc tttacataca tgctggagaa
     1981 gtggaggtgg atggtattta agggggagat accgaaagat caatggatga aaaagtggtg
     2041 ggaaatgaaa agggagatcg ttggcgtagt tgaaccagta ccgcatgatg agacgtactg
     2101 cgatccggct agtctgttcc atgtctctaa tgattactct ttcatccgct actacacccg
     2161 cacgctgtat caattccagt tccaagaagc tctctgtcag gctgccaagc acgaaggacc
     2221 gctgcacaaa tgcgacatta gcaattctac agaggcgggt cagaagttgt tcaatatgct
     2281 tagactgggg aagagcgaac cgtggacgct cgctttggag aacgttgttg gagctaagaa
     2341 tatgaacgtc aggcccttgc tgaattactt tgaacctctg tttacgtggt tgaaagacca
     2401 aaataaaaac tcctttgttg ggtggagtac tgactggtcc ccctatgcgg accaaagcat
     2461 caaagtgagg ataagcctaa aatcagctct tggagataaa gcatatgaat ggaacgacaa
     2521 tgaaatgtac ctgttccgat catctgttgc atatgctatg aggcagtact ttttaaaagt
     2581 aaaaaatcag atgattcttt ttggggagga ggatgtgcga gtggctaatt tgaaaccaag
     2641 aatctccttt aatttctttg tcactgcacc taaaaatgtg tctgatatca ttcctagaac
     2701 tgaagttgaa aaggccatca ggatgtcccg gagccgtatc aatgatgctt tccgtctgaa
     2761 tgacaacagc ctagagtttc tggggataca gccaacactt ggacctccta accagccccc
     2821 tgtttccact agttctggtg gtggtggtga gaatctgtac tttcagagct cgggcggagg
     2881 atcgggtgga ggcgagccca aatcttgtga caaaactcac acatgccctc catgcccagc
     2941 gccagaattg ctgggcggac ccagcgtgtt cctgttccca cctaaaccta aagacaccct
     3001 gatgatcagc cgaacccctg aggtgacctg cgtggtggtg gacgtgagcc acgaggaccc
     3061 cgaggtgaag ttcaactggt atgtggacgg cgtggaggtc cacaatgcca aaacgaagcc
     3121 cagggaggag cagtacaaca gcacctacag ggtagtgagc gtcttgaccg tgctgcacca
     3181 ggactggctg aacggcaagg aatacaaatg caaggtcagc aataaggctc tgccggctcc
     3241 tatcgagaag acaatcagca aggcaaaggg ccagccacgc gaaccgcagg tgtatactct
     3301 gccccccagc cgggacgagc tgaccaagaa ccaggtgtcc ctgacctgtc tggtgaaagg
     3361 cttctacccc agcgacatcg ctgtggagtg ggagagtaac gggcagcccg agaacaacta
     3421 caagaccacg cctcctgtgc tggacagcga cggcagcttc ttcctgtata gcaagctcac
     3481 cgtggacaag agcaggtggc aacagggcaa cgtgttcagc tgctctgtga tgcacgaggc
     3541 cctgcacaac cattacaccc agaagagtct cagtctgagc ccgggaaagg gtggaggcgg
     3601 atccggcctg aacgacatct tcgaggctca gaaaatcgaa tggcacgaag gctaatccca
     3661 gtgtcgctag ctggccagac atgataagat acattgatga gtttggacaa accacaacta
     3721 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3781 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3841 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     3901 aattaattct aaaatacagc atagcaaaac tttaacctcc aaatcaagcc tctacttgaa
     3961 tccttttctg agggatgaat aaggcatagg catcaggggc tgttgccaat gtgcattagc
     4021 tgtttgcagc ctcaccttct ttcatggagt ttaagatata gtgtattttc ccaaggtttg
     4081 aactagctct tcatttcttt atgttttaaa tgcactgacc tcccacattc cctttttagt
     4141 aaaatattca gaaataattt aaatacatca ttgcaatgaa aataaatgtt ttttattagg
     4201 cagaatccag atgctcaagg cccttcataa tatcccccag tttagtagtt ggacttaggg
     4261 aacaaaggaa cctttaatag aaattggaca gcaagaaagc gagcttctag cttatcctca
     4321 gtcctgctcc tctgccacaa agtgcacgca gttgccggcc gggtcgcgca gggcgaactc
     4381 ccgcccccac ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt
     4441 cgtggacacg acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca
     4501 ggccagggtg ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc
     4561 gtcccggacc acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc
     4621 ggtccagaac tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt
     4681 caacttggcc atgatggctc ctcctgtcag gagaggaaag agaagaaggt tagtacaatt
     4741 gctatagtga gttgtattat actatgcaga tatactatgc caatgattaa ttgtcaaact
     4801 agggctgcag ggttcatagt gccacttttc ctgcactgcc ccatctcctg cccacccttt
     4861 cccaggcata gacagtcagt gacttaccaa actcacagga gggagaaggc agaagcttga
     4921 gacagacccg cgggaccgcc gaactgcgag gggacgtggc tagggcggct tcttttatgg
     4981 tgcgccggcc ctcggaggca gggcgctcgg ggaggcctag cggccaatct gcggtggcag
     5041 gaggcggggc cgaaggccgt gcctgaccaa tccggagcac ataggagtct cagccccccg
     5101 ccccaaagca aggggaagtc acgcgcctgt agcgccagcg tgttgtgaaa tgggggcttg
     5161 ggggggttgg ggccctgatt agtcaaaaca aactcccatt gacgtcaatg gggtggagac
     5221 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     5281 tcatcatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     5341 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     5401 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     5461 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     5521 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     5581 atgtaacgcc tgcaggttaa ttaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5641 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5701 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5761 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5821 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5881 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5941 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6001 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6061 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6121 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6181 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6241 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6301 cgaaaactca cgttaaggga ttttggtcat ggctagttaa ttaacattta aatcagcggc
     6361 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgtaa
     6421 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
     6481 cccagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     6541 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     6601 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     6661 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     6721 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     6781 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     6841 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     6901 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     6961 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     7021 aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg
     7081 aaattaatac gactcactat agggagaccc aagctggcta gttaagcttg ccaccatgaa
     7141 ggataacacc gtgccactga aattgattgc cctgttagcg aacggtgaat ttcactctgg
     7201 cgagcagttg ggtgaaacgc tgggaatgag ccgggcggct attaataaac acattcagac
     7261 actgcgtgac tggggcgttg atgtctttac cgttccgggt aaaggataca gcctgcctga
     7321 gcctatccag ttacttaatg ctaaacagat attgggtcag ctggatggcg gtagtgtagc
     7381 cgtgctgcca gtgattgact ccacgaatca gtaccttctt gatcgtatcg gagagcttaa
     7441 atcgggcgat gcttgcattg cagaatacca gcaggctggc cgtggtcgcc ggggtcggaa
     7501 atggttttcg ccttttggcg caaacttata tttgtcgatg ttctggcgtc tggaacaagg
     7561 cccggcggcg gcgattggtt taagtctggt tatcggtatc gtgatggcgg aagtattacg
     7621 caagctgggt gcagataaag ttcgtgttaa atggcctaat gacctctatc tgcaggatcg
     7681 caagctggca ggcattctgg tggagctgac tggcaaaact ggcgatgcgg cgcaaatagt
     7741 cattggagcc gggatcaaca tggcaatgcg ccgtgttgaa gagagtgtcg ttaatcaggg
     7801 gtggatcacg ctgcaggaag cggggatcaa tctcgatcgt aatacgttgg cggccatgct
     7861 aatacgtgaa ttacgtgctg cgttggaact cttcgaacaa gaaggattgg caccttatct
     7921 gtcgcgctgg gaaaagctgg ataattttat taatcgccca gtgaaactta tcattggtga
     7981 taaagaaata tttggcattt cacgcggaat agacaaacag ggggctttat tacttgagca
     8041 ggatggaata ataaaaccct ggatgggcgg tgaaatatcc ctgcgtagtg cagaaaaata
     8101 atctagaggg cccgcggttc gaacaaaaac tcatctcaga agaggatctg aatatgcata
     8161 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     8221 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     8281 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     8341 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     8401 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcgggt gcaagtgcag
     8461 g
//
