LOCUS       sequence-314502-        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: pCVD313
FEATURES             Location/Qualifiers
     source          1..8461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..251
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             264..532
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2835..2855
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3591..3635
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3671..3792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3682..3701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3736..3755
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    3896..4290
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4304..4678)
                     /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        5271..5574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5654..6242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5734..5753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6350..6398
                     /note="synthetic polyadenylation signal"
     primer_bind     6449..6468
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        6519..6822
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6823..7026
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6977..6997
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7071..7090
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7071..7089
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7122..8087
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             7122..8084
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             8107..8136
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             8152..8169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8192..8209)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8198..8422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 tctatcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccaa
       61 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
      121 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
      181 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
      241 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
      301 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
      361 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
      421 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
      481 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
      541 gaccggcgcc tacctgagat caccggcgaa ggagggccac catgcgaatg cagctgctgc
      601 tgctgattgc gctgagcctg gcgctggtga ccaacagcac tagtcaatct accatcgaag
      661 agcaggccaa aacattcctc gacttctttg atagccaggc tgaagacctt ttctaccaat
      721 caagtctggc tagctggaat tacaatacaa acattacaga ggagaacgta caaaacatga
      781 ataacgcagg ggacaagtgg agcgcattcc ttaaggaaca aagtaccctt gcgcaaatgt
      841 atccgctgca agagattcaa aacctgacgg ttaagctgca acttcaggcc ctccaacaaa
      901 atggatcctc agtcttgtca gaagacaaaa gcaagcgact gaacaccatc cttaacacca
      961 tgtcaaccat atattcaaca ggtaaagttt gcaatccgga taacccccaa gaatgtttgc
     1021 ttcttgaacc cggtctcaac gaaattatgg ccaacagtct tgattacaac gagcgattgt
     1081 gggcatggga aagttggagg agtgaggtag gcaaacagtt gagacctctt tatgaagagt
     1141 acgttgtcct taaaaatgaa atggctcgcg cgaatcatta tgaagactat ggtgactact
     1201 ggagggggga ttatgaggtg aacggggtgg acggatacga ttactctagg ggccagctga
     1261 tagaggatgt cgagcacacc tttgaggaga ttaagccgtt gtacgaacat ttgcacgcct
     1321 atgtcagggc taagctcatg aacgcttatc cgagttatat ctccccgata ggatgcttgc
     1381 ctgctcactt gttgggcgat atgtggggac gcttttggac caacttgtat tcccttacgg
     1441 taccgttcgg ccagaaacca aatatcgacg tgacagacgc aatggtggat caagcatggg
     1501 atgcgcaacg aatcttcaag gaggcagaaa aatttttcgt ttcagttgga ctcccaaaca
     1561 tgacgcaggg tttctgggag aactcaatgt tgacagatcc aggtaatgtg cagaaagcgg
     1621 tttgcctccc tactgcatgg gatcttggta aaggggactt ccgcatactc atgtgtacga
     1681 aagtaactat ggacgacttt cttactgcgc accacgagat ggggcacata caatacgata
     1741 tggcgtacgc agctcaacct ttccttctgc ggaacggggc gaatgaagga tttcacgagg
     1801 cagtgggtga gattatgtcc ctgtcagctg ccactccgaa acatctgaaa agcatcggcc
     1861 tgttgagccc agacttccaa gaagataatg agaccgaaat aaacttcctt ctgaagcaag
     1921 cactgactat tgtaggtacc ttgcccttta catacatgct ggagaagtgg aggtggatgg
     1981 tatttaaggg ggagataccg aaagatcaat ggatgaaaaa gtggtgggaa atgaaaaggg
     2041 agatcgttgg cgtagttgaa ccagtaccgc atgatgagac gtactgcgat ccggctagtc
     2101 tgttccatgt ctctaatgat tactctttca tccgctacta cacccgcacg ctgtatcaat
     2161 tccagttcca agaagctctc tgtcaggctg ccaagcacga aggaccgctg cacaaatgcg
     2221 acattagcaa ttctacagag gcgggtcaga agttgttcaa tatgcttaga ctggggaaga
     2281 gcgaaccgtg gacgctcgct ttggagaacg ttgttggagc taagaatatg aacgtcaggc
     2341 ccttgctgaa ttactttgaa cctctgttta cgtggttgaa agaccaaaat aaaaactcct
     2401 ttgttgggtg gagtactgac tggtccccct atgcggacca aagcatcaaa gtgaggataa
     2461 gcctaaaatc agctcttgga gataaagcat atgaatggaa cgacaatgaa atgtacctgt
     2521 tccgatcatc tgttgcatat gctatgaggc agtacttttt aaaagtaaaa aatcagatga
     2581 ttctttttgg ggaggaggat gtgcgagtgg ctaatttgaa accaagaatc tcctttaatt
     2641 tctttgtcac tgcacctaaa aatgtgtctg atatcattcc tagaactgaa gttgaaaagg
     2701 ccatcaggat gtcccggagc cgtatcaatg atgctttccg tctgaatgac aacagcctag
     2761 agtttctggg gatacagcca acacttggac ctcctaacca gccccctgtt tccactagtt
     2821 ctggtggtgg tggtgagaat ctgtactttc agagctcggg cggaggatcg ggtggaggcg
     2881 agcccaaatc ttgtgacaaa actcacacat gccccccctg cccagcgcca gaattgctgg
     2941 gcggacccag cgtgttcctg ttccccccca aacctaaaga caccctgatg atcagccgaa
     3001 cccctgaggt gacctgcgtg gtggtggacg tgagccacga ggaccccgag gtgaagttca
     3061 actggtatgt ggacggcgtg gaggtccaca atgccaaaac gaagcccagg gaggagcagt
     3121 acaacagcac ctacagggta gtgagcgtct tgaccgtgct gcaccaggac tggctgaacg
     3181 gcaaggaata caaatgcaag gtcagcaata aggctctgcc ggctcctatc gagaagacaa
     3241 tcagcaaggc aaagggccag ccacgcgaac cgcaggtgta tactctgccc cccagccggg
     3301 acgagctgac caagaaccag gtgtccctga cctgtctggt gaaaggcttc taccccagcg
     3361 acatcgctgt ggagtgggag agtaacgggc agcccgagaa caactacaag accacgcctc
     3421 ctgtgctgga cagcgacggc agcttcttcc tgtatagcaa gctcaccgtg gacaagagca
     3481 ggtggcaaca gggcaacgtg ttcagctgct ctgtgatgca cgaggccctg cacaaccatt
     3541 acacccagaa gagtctcagt ctgagcccgg gaaagggtgg aggcggatcc ggcctgaacg
     3601 acatcttcga ggctcagaaa atcgaatggc acgaaggcta atcccagtgt cgctagctgg
     3661 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3721 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3781 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3841 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggaatt aattctaaaa
     3901 tacagcatag caaaacttta acctccaaat caagcctcta cttgaatcct tttctgaggg
     3961 atgaataagg cataggcatc aggggctgtt gccaatgtgc attagctgtt tgcagcctca
     4021 ccttctttca tggagtttaa gatatagtgt attttcccaa ggtttgaact agctcttcat
     4081 ttctttatgt tttaaatgca ctgacctccc acattccctt tttagtaaaa tattcagaaa
     4141 taatttaaat acatcattgc aatgaaaata aatgtttttt attaggcaga atccagatgc
     4201 tcaaggccct tcataatatc ccccagttta gtagttggac ttagggaaca aaggaacctt
     4261 taatagaaat tggacagcaa gaaagcgagc ttctagctta tcctcagtcc tgctcctctg
     4321 ccacaaagtg cacgcagttg ccggccgggt cgcgcagggc gaactcccgc ccccacggct
     4381 gctcgccgat ctcggtcatg gccggcccgg aggcgtcccg gaagttcgtg gacacgacct
     4441 ccgaccactc ggcgtacagc tcgtccaggc cgcgcaccca cacccaggcc agggtgttgt
     4501 ccggcaccac ctggtcctgg accgcgctga tgaacagggt cacgtcgtcc cggaccacac
     4561 cggcgaagtc gtcctccacg aagtcccggg agaacccgag ccggtcggtc cagaactcga
     4621 ccgctccggc gacgtcgcgc gcggtgagca ccggaacggc actggtcaac ttggccatga
     4681 tggctcctcc tgtcaggaga ggaaagagaa gaaggttagt acaattgcta tagtgagttg
     4741 tattatacta tgcagatata ctatgccaat gattaattgt caaactaggg ctgcagggtt
     4801 catagtgcca cttttcctgc actgccccat ctcctgccca ccctttccca ggcatagaca
     4861 gtcagtgact taccaaactc acaggaggga gaaggcagaa gcttgagaca gacccgcggg
     4921 accgccgaac tgcgagggga cgtggctagg gcggcttctt ttatggtgcg ccggccctcg
     4981 gaggcagggc gctcggggag gcctagcggc caatctgcgg tggcaggagg cggggccgaa
     5041 ggccgtgcct gaccaatccg gagcacatag gagtctcagc cccccgcccc aaagcaaggg
     5101 gaagtcacgc gcctgtagcg ccagcgtgtt gtgaaatggg ggcttggggg ggttggggcc
     5161 ctgattagtc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     5221 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcat catggtaata
     5281 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     5341 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     5401 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     5461 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     5521 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcctgca
     5581 ggttaattaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5641 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5701 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5761 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5821 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5881 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5941 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6001 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6061 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6121 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6181 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6301 aagggatttt ggtcatggct agttaattaa catttaaatc agcggccgca ataaaatatc
     6361 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgtaactaa catacgctct
     6421 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtaatcaatt
     6481 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6541 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6601 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6661 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     6721 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     6781 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     6841 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     6901 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     6961 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     7021 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     7081 cactataggg agacccaagc tggctagtta agcttgccac catgaaggat aacaccgtgc
     7141 cactgaaatt gattgccctg ttagcgaacg gtgaatttca ctctggcgag cagttgggtg
     7201 aaacgctggg aatgagccgg gcggctatta ataaacacat tcagacactg cgtgactggg
     7261 gcgttgatgt ctttaccgtt ccgggtaaag gatacagcct gcctgagcct atccagttac
     7321 ttaatgctaa acagatattg ggtcagctgg atggcggtag tgtagccgtg ctgccagtga
     7381 ttgactccac gaatcagtac cttcttgatc gtatcggaga gcttaaatcg ggcgatgctt
     7441 gcattgcaga ataccagcag gctggccgtg gtcgccgggg tcggaaatgg ttttcgcctt
     7501 ttggcgcaaa cttatatttg tcgatgttct ggcgtctgga acaaggcccg gcggcggcga
     7561 ttggtttaag tctggttatc ggtatcgtga tggcggaagt attacgcaag ctgggtgcag
     7621 ataaagttcg tgttaaatgg cctaatgacc tctatctgca ggatcgcaag ctggcaggca
     7681 ttctggtgga gctgactggc aaaactggcg atgcggcgca aatagtcatt ggagccggga
     7741 tcaacatggc aatgcgccgt gttgaagaga gtgtcgttaa tcaggggtgg atcacgctgc
     7801 aggaagcggg gatcaatctc gatcgtaata cgttggcggc catgctaata cgtgaattac
     7861 gtgctgcgtt ggaactcttc gaacaagaag gattggcacc ttatctgtcg cgctgggaaa
     7921 agctggataa ttttattaat cgcccagtga aacttatcat tggtgataaa gaaatatttg
     7981 gcatttcacg cggaatagac aaacaggggg ctttattact tgagcaggat ggaataataa
     8041 aaccctggat gggcggtgaa atatccctgc gtagtgcaga aaaataatct agagggcccg
     8101 cggttcgaac aaaaactcat ctcagaagag gatctgaata tgcataccgg tcatcatcac
     8161 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     8221 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     8281 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     8341 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     8401 ggggatgcgg tgggctctat ggcttctgag gcgggtgcaa gtgcaggtgc cagaacattt
     8461 c
//
