LOCUS       sequence-314499-        8083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for wild-type Fc-fused ACE2(614).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8083)
  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 8083)
  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: pCVD013
FEATURES             Location/Qualifiers
     source          1..8083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     515..535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     609..628
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        609..627
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             660..1625
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             660..1622
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1645..1674
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1690..1707
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1730..1747)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1736..1960
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2039..2250
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2263..2531
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4456..4476
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5212..5256
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    5292..5413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5303..5322)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5357..5376
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    5517..5911
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(5925..6299)
                     /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        6892..7195
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(7275..7863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7355..7374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7971..8019
                     /note="synthetic polyadenylation signal"
     primer_bind     join(8070..8083,1..6)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 tgtccccagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      541 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      601 atcgaaatta atacgactca ctatagggag acccaagctg gctagttaag cttgccacca
      661 tgaaggataa caccgtgcca ctgaaattga ttgccctgtt agcgaacggt gaatttcact
      721 ctggcgagca gttgggtgaa acgctgggaa tgagccgggc ggctattaat aaacacattc
      781 agacactgcg tgactggggc gttgatgtct ttaccgttcc gggtaaagga tacagcctgc
      841 ctgagcctat ccagttactt aatgctaaac agatattggg tcagctggat ggcggtagtg
      901 tagccgtgct gccagtgatt gactccacga atcagtacct tcttgatcgt atcggagagc
      961 ttaaatcggg cgatgcttgc attgcagaat accagcaggc tggccgtggt cgccggggtc
     1021 ggaaatggtt ttcgcctttt ggcgcaaact tatatttgtc gatgttctgg cgtctggaac
     1081 aaggcccggc ggcggcgatt ggtttaagtc tggttatcgg tatcgtgatg gcggaagtat
     1141 tacgcaagct gggtgcagat aaagttcgtg ttaaatggcc taatgacctc tatctgcagg
     1201 atcgcaagct ggcaggcatt ctggtggagc tgactggcaa aactggcgat gcggcgcaaa
     1261 tagtcattgg agccgggatc aacatggcaa tgcgccgtgt tgaagagagt gtcgttaatc
     1321 aggggtggat cacgctgcag gaagcgggga tcaatctcga tcgtaatacg ttggcggcca
     1381 tgctaatacg tgaattacgt gctgcgttgg aactcttcga acaagaagga ttggcacctt
     1441 atctgtcgcg ctgggaaaag ctggataatt ttattaatcg cccagtgaaa cttatcattg
     1501 gtgataaaga aatatttggc atttcacgcg gaatagacaa acagggggct ttattacttg
     1561 agcaggatgg aataataaaa ccctggatgg gcggtgaaat atccctgcgt agtgcagaaa
     1621 aataatctag agggcccgcg gttcgaacaa aaactcatct cagaagagga tctgaatatg
     1681 cataccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg
     1741 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1801 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1861 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1921 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc gggtgcaagt
     1981 gcaggtgcca gaacatttct ctatcgaagg atctgcgatc gctccggtgc ccgtcagtgg
     2041 gcagagcgca catcgccaac agtccccgag aagttggggg gaggggtcgg caattgaacg
     2101 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     2161 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     2221 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     2281 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     2341 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     2401 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     2461 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     2521 cgttacagat ccaagctgtg accggcgcct acctgagatc accggcgaag gagggccacc
     2581 atgcgaatgc agctgctgct gctgattgcg ctgagcctgg cgctggtgac caacagcact
     2641 agtcaatcta ccatcgaaga gcaggccaaa acattcctcg acaagtttaa tcacgaggct
     2701 gaagaccttt tctaccaatc aagtctggct agctggaatt acaatacaaa cattacagag
     2761 gagaacgtac aaaacatgaa taacgcaggg gacaagtgga gcgcattcct taaggaacaa
     2821 agtacccttg cgcaaatgta tccgctgcaa gagattcaaa acctgacggt taagctgcaa
     2881 cttcaggccc tccaacaaaa tggaagttca gtcttgtcag aagacaaaag caagcgactg
     2941 aacaccatcc ttaacaccat gtcaaccata tattcaacag gtaaagtttg caatccggat
     3001 aacccccaag aatgtttgct tcttgaaccc ggtctcaacg aaattatggc caacagtctt
     3061 gattacaacg agcgattgtg ggcatgggaa agttggagga gtgaggtagg caaacagttg
     3121 agacctcttt atgaagagta cgttgtcctt aaaaatgaaa tggctcgcgc gaatcattat
     3181 gaagactatg gtgactactg gaggggggat tatgaggtga acggggtgga cggatacgat
     3241 tactctaggg gccagctgat agaggatgtc gagcacacct ttgaggagat taagccgttg
     3301 tacgaacatt tgcacgccta tgtcagggct aagctcatga acgcttatcc gagttatatc
     3361 tccccgatag gatgcttgcc tgctcacttg ttgggcgata tgtggggacg cttttggacc
     3421 aacttgtatt cccttacggt accgttcggc cagaaaccaa atatcgacgt gacagacgca
     3481 atggtggatc aagcatggga tgcgcaacga atcttcaagg aggcagaaaa atttttcgtt
     3541 tcagttggac tcccaaacat gacgcagggt ttctgggaga actcaatgtt gacagatcca
     3601 ggtaatgtgc agaaagcggt ttgccaccct actgcatggg atcttggtaa aggggacttc
     3661 cgcatactca tgtgtacgaa agtaactatg gacgactttc ttactgcgca ccacgagatg
     3721 gggcacatac aatacgatat ggcgtacgca gctcaacctt tccttctgcg gaacggggcg
     3781 aatgaaggat ttcacgaggc agtgggtgag attatgtccc tgtcagctgc cactccgaaa
     3841 catctgaaaa gcatcggcct gttgagccca gacttccaag aagataatga gaccgaaata
     3901 aacttccttc tgaagcaagc actgactatt gtaggtacct tgccctttac atacatgctg
     3961 gagaagtgga ggtggatggt atttaagggg gagataccga aagatcaatg gatgaaaaag
     4021 tggtgggaaa tgaaaaggga gatcgttggc gtagttgaac cagtaccgca tgatgagacg
     4081 tactgcgatc cggctagtct gttccatgtc tctaatgatt actctttcat ccgctactac
     4141 acccgcacgc tgtatcaatt ccagttccaa gaagctctct gtcaggctgc caagcacgaa
     4201 ggaccgctgc acaaatgcga cattagcaat tctacagagg cgggtcagaa gttgttcaat
     4261 atgcttagac tggggaagag cgaaccgtgg acgctcgctt tggagaacgt tgttggagct
     4321 aagaatatga acgtcaggcc cttgctgaat tactttgaac ctctgtttac gtggttgaaa
     4381 gaccaaaata aaaactcctt tgttgggtgg agtactgact ggtcccccta tgcgactagt
     4441 tctggtggtg gtggtgagaa tctgtacttt cagagctcgg gcggaggatc gggtggaggc
     4501 gagcccaaat cttgtgacaa aactcacaca tgccccccct gcccagcgcc agaattgctg
     4561 ggcggaccca gcgtgttcct gttccccccc aaacctaaag acaccctgat gatcagccga
     4621 acccctgagg tgacctgcgt ggtggtggac gtgagccacg aggaccccga ggtgaagttc
     4681 aactggtatg tggacggcgt ggaggtccac aatgccaaaa cgaagcccag ggaggagcag
     4741 tacaacagca cctacagggt agtgagcgtc ttgaccgtgc tgcaccagga ctggctgaac
     4801 ggcaaggaat acaaatgcaa ggtcagcaat aaggctctgc cggctcctat cgagaagaca
     4861 atcagcaagg caaagggcca gccacgcgaa ccgcaggtgt atactctgcc ccccagccgg
     4921 gacgagctga ccaagaacca ggtgtccctg acctgtctgg tgaaaggctt ctaccccagc
     4981 gacatcgctg tggagtggga gagtaacggg cagcccgaga acaactacaa gaccacgcct
     5041 cctgtgctgg acagcgacgg cagcttcttc ctgtatagca agctcaccgt ggacaagagc
     5101 aggtggcaac agggcaacgt gttcagctgc tctgtgatgc acgaggccct gcacaaccat
     5161 tacacccaga agagtctcag tctgagcccg ggaaagggtg gaggcggatc cggcctgaac
     5221 gacatcttcg aggctcagaa aatcgaatgg cacgaaggct aatcccagtg tcgctagctg
     5281 gccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     5341 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     5401 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     5461 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggaat taattctaaa
     5521 atacagcata gcaaaacttt aacctccaaa tcaagcctct acttgaatcc ttttctgagg
     5581 gatgaataag gcataggcat caggggctgt tgccaatgtg cattagctgt ttgcagcctc
     5641 accttctttc atggagttta agatatagtg tattttccca aggtttgaac tagctcttca
     5701 tttctttatg ttttaaatgc actgacctcc cacattccct ttttagtaaa atattcagaa
     5761 ataatttaaa tacatcattg caatgaaaat aaatgttttt tattaggcag aatccagatg
     5821 ctcaaggccc ttcataatat cccccagttt agtagttgga cttagggaac aaaggaacct
     5881 ttaatagaaa ttggacagca agaaagcgag cttctagctt atcctcagtc ctgctcctct
     5941 gccacaaagt gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg cccccacggc
     6001 tgctcgccga tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc
     6061 tccgaccact cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg
     6121 tccggcacca cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca
     6181 ccggcgaagt cgtcctccac gaagtcccgg gagaacccga gccggtcggt ccagaactcg
     6241 accgctccgg cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa cttggccatg
     6301 atggctcctc ctgtcaggag aggaaagaga agaaggttag tacaattgct atagtgagtt
     6361 gtattatact atgcagatat actatgccaa tgattaattg tcaaactagg gctgcagggt
     6421 tcatagtgcc acttttcctg cactgcccca tctcctgccc accctttccc aggcatagac
     6481 agtcagtgac ttaccaaact cacaggaggg agaaggcaga agcttgagac agacccgcgg
     6541 gaccgccgaa ctgcgagggg acgtggctag ggcggcttct tttatggtgc gccggccctc
     6601 ggaggcaggg cgctcgggga ggcctagcgg ccaatctgcg gtggcaggag gcggggccga
     6661 aggccgtgcc tgaccaatcc ggagcacata ggagtctcag ccccccgccc caaagcaagg
     6721 ggaagtcacg cgcctgtagc gccagcgtgt tgtgaaatgg gggcttgggg gggttggggc
     6781 cctgattagt caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     6841 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca tcatggtaat
     6901 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     6961 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     7021 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     7081 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     7141 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcctgc
     7201 aggttaatta agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7441 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7681 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7801 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7861 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7921 taagggattt tggtcatggc tagttaatta acatttaaat cagcggccgc aataaaatat
     7981 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgtaacta acatacgctc
     8041 tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggc
//
