LOCUS       sequence-314497-        8461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for wild-type 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: pCVD208
FEATURES             Location/Qualifiers
     source          1..8461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     621..640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             672..1637
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             672..1634
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1657..1686
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1702..1719
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1742..1759)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1748..1972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2051..2262
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2275..2543
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4846..4866
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5602..5646
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    5682..5803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5693..5712)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5747..5766
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    5907..6301
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(6315..6689)
                     /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        7282..7585
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(7665..8253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7745..7764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    8361..8409
                     /note="synthetic polyadenylation signal"
     primer_bind     join(8460..8461,1..18)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 agcaaaatag gctgtcccca gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      601 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagtta
      661 agcttgccac catgaaggat aacaccgtgc cactgaaatt gattgccctg ttagcgaacg
      721 gtgaatttca ctctggcgag cagttgggtg aaacgctggg aatgagccgg gcggctatta
      781 ataaacacat tcagacactg cgtgactggg gcgttgatgt ctttaccgtt ccgggtaaag
      841 gatacagcct gcctgagcct atccagttac ttaatgctaa acagatattg ggtcagctgg
      901 atggcggtag tgtagccgtg ctgccagtga ttgactccac gaatcagtac cttcttgatc
      961 gtatcggaga gcttaaatcg ggcgatgctt gcattgcaga ataccagcag gctggccgtg
     1021 gtcgccgggg tcggaaatgg ttttcgcctt ttggcgcaaa cttatatttg tcgatgttct
     1081 ggcgtctgga acaaggcccg gcggcggcga ttggtttaag tctggttatc ggtatcgtga
     1141 tggcggaagt attacgcaag ctgggtgcag ataaagttcg tgttaaatgg cctaatgacc
     1201 tctatctgca ggatcgcaag ctggcaggca ttctggtgga gctgactggc aaaactggcg
     1261 atgcggcgca aatagtcatt ggagccggga tcaacatggc aatgcgccgt gttgaagaga
     1321 gtgtcgttaa tcaggggtgg atcacgctgc aggaagcggg gatcaatctc gatcgtaata
     1381 cgttggcggc catgctaata cgtgaattac gtgctgcgtt ggaactcttc gaacaagaag
     1441 gattggcacc ttatctgtcg cgctgggaaa agctggataa ttttattaat cgcccagtga
     1501 aacttatcat tggtgataaa gaaatatttg gcatttcacg cggaatagac aaacaggggg
     1561 ctttattact tgagcaggat ggaataataa aaccctggat gggcggtgaa atatccctgc
     1621 gtagtgcaga aaaataatct agagggcccg cggttcgaac aaaaactcat ctcagaagag
     1681 gatctgaata tgcataccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     1741 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1801 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1861 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1921 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     1981 gcgggtgcaa gtgcaggtgc cagaacattt ctctatcgaa ggatctgcga tcgctccggt
     2041 gcccgtcagt gggcagagcg cacatcgcca acagtccccg agaagttggg gggaggggtc
     2101 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2161 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2221 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     2281 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     2341 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     2401 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     2461 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     2521 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacctgaga tcaccggcga
     2581 aggagggcca ccatgcgaat gcagctgctg ctgctgattg cgctgagcct ggcgctggtg
     2641 accaacagca ctagtcaatc taccatcgaa gagcaggcca aaacattcct cgacaagttt
     2701 aatcacgagg ctgaagacct tttctaccaa tcaagtctgg ctagctggaa ttacaataca
     2761 aacattacag aggagaacgt acaaaacatg aataacgcag gggacaagtg gagcgcattc
     2821 cttaaggaac aaagtaccct tgcgcaaatg tatccgctgc aagagattca aaacctgacg
     2881 gttaagctgc aacttcaggc cctccaacaa aatggaagtt cagtcttgtc agaagacaaa
     2941 agcaagcgac tgaacaccat ccttaacacc atgtcaacca tatattcaac aggtaaagtt
     3001 tgcaatccgg ataaccccca agaatgtttg cttcttgaac ccggtctcaa cgaaattatg
     3061 gccaacagtc ttgattacaa cgagcgattg tgggcatggg aaagttggag gagtgaggta
     3121 ggcaaacagt tgagacctct ttatgaagag tacgttgtcc ttaaaaatga aatggctcgc
     3181 gcgaatcatt atgaagacta tggtgactac tggagggggg attatgaggt gaacggggtg
     3241 gacggatacg attactctag gggccagctg atagaggatg tcgagcacac ctttgaggag
     3301 attaagccgt tgtacgaaca tttgcacgcc tatgtcaggg ctaagctcat gaacgcttat
     3361 ccgagttata tctccccgat aggatgcttg cctgctcact tgttgggcga tatgtgggga
     3421 cgcttttgga ccaacttgta ttcccttacg gtaccgttcg gccagaaacc aaatatcgac
     3481 gtgacagacg caatggtgga tcaagcatgg gatgcgcaac gaatcttcaa ggaggcagaa
     3541 aaatttttcg tttcagttgg actcccaaac atgacgcagg gtttctggga gaactcaatg
     3601 ttgacagatc caggtaatgt gcagaaagcg gtttgccacc ctactgcatg ggatcttggt
     3661 aaaggggact tccgcatact catgtgtacg aaagtaacta tggacgactt tcttactgcg
     3721 caccacgaga tggggcacat acaatacgat atggcgtacg cagctcaacc tttccttctg
     3781 cggaacgggg cgaatgaagg atttcacgag gcagtgggtg agattatgtc cctgtcagct
     3841 gccactccga aacatctgaa aagcatcggc ctgttgagcc cagacttcca agaagataat
     3901 gagaccgaaa taaacttcct tctgaagcaa gcactgacta ttgtaggtac cttgcccttt
     3961 acatacatgc tggagaagtg gaggtggatg gtatttaagg gggagatacc gaaagatcaa
     4021 tggatgaaaa agtggtggga aatgaaaagg gagatcgttg gcgtagttga accagtaccg
     4081 catgatgaga cgtactgcga tccggctagt ctgttccatg tctctaatga ttactctttc
     4141 atccgctact acacccgcac gctgtatcaa ttccagttcc aagaagctct ctgtcaggct
     4201 gccaagcacg aaggaccgct gcacaaatgc gacattagca attctacaga ggcgggtcag
     4261 aagttgttca atatgcttag actggggaag agcgaaccgt ggacgctcgc tttggagaac
     4321 gttgttggag ctaagaatat gaacgtcagg cccttgctga attactttga acctctgttt
     4381 acgtggttga aagaccaaaa taaaaactcc tttgttgggt ggagtactga ctggtccccc
     4441 tatgcggacc aaagcatcaa agtgaggata agcctaaaat cagctcttgg agataaagca
     4501 tatgaatgga acgacaatga aatgtacctg ttccgatcat ctgttgcata tgctatgagg
     4561 cagtactttt taaaagtaaa aaatcagatg attctttttg gggaggagga tgtgcgagtg
     4621 gctaatttga aaccaagaat ctcctttaat ttctttgtca ctgcacctaa aaatgtgtct
     4681 gatatcattc ctagaactga agttgaaaag gccatcagga tgtcccggag ccgtatcaat
     4741 gatgctttcc gtctgaatga caacagccta gagtttctgg ggatacagcc aacacttgga
     4801 cctcctaacc agccccctgt ttccactagt tctggtggtg gtggtgagaa tctgtacttt
     4861 cagagctcgg gcggaggatc gggtggaggc gagcccaaat cttgtgacaa aactcacaca
     4921 tgccctccat gcccagcgcc agaattgctg ggcggaccca gcgtgttcct gttcccacct
     4981 aaacctaaag acaccctgat gatcagccga acccctgagg tgacctgcgt ggtggtggac
     5041 gtgagccacg aggaccccga ggtgaagttc aactggtatg tggacggcgt ggaggtccac
     5101 aatgccaaaa cgaagcccag ggaggagcag tacaacagca cctacagggt agtgagcgtc
     5161 ttgaccgtgc tgcaccagga ctggctgaac ggcaaggaat acaaatgcaa ggtcagcaat
     5221 aaggctctgc cggctcctat cgagaagaca atcagcaagg caaagggcca gccacgcgaa
     5281 ccgcaggtgt atactctgcc ccccagccgg gacgagctga ccaagaacca ggtgtccctg
     5341 acctgtctgg tgaaaggctt ctaccccagc gacatcgctg tggagtggga gagtaacggg
     5401 cagcccgaga acaactacaa gaccacgcct cctgtgctgg acagcgacgg cagcttcttc
     5461 ctgtatagca agctcaccgt ggacaagagc aggtggcaac agggcaacgt gttcagctgc
     5521 tctgtgatgc acgaggccct gcacaaccat tacacccaga agagtctcag tctgagcccg
     5581 ggaaagggtg gaggcggatc cggcctgaac gacatcttcg aggctcagaa aatcgaatgg
     5641 cacgaaggct aatcccagtg tcgctagctg gccagacatg ataagataca ttgatgagtt
     5701 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     5761 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     5821 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     5881 ctacaaatgt ggtatggaat taattctaaa atacagcata gcaaaacttt aacctccaaa
     5941 tcaagcctct acttgaatcc ttttctgagg gatgaataag gcataggcat caggggctgt
     6001 tgccaatgtg cattagctgt ttgcagcctc accttctttc atggagttta agatatagtg
     6061 tattttccca aggtttgaac tagctcttca tttctttatg ttttaaatgc actgacctcc
     6121 cacattccct ttttagtaaa atattcagaa ataatttaaa tacatcattg caatgaaaat
     6181 aaatgttttt tattaggcag aatccagatg ctcaaggccc ttcataatat cccccagttt
     6241 agtagttgga cttagggaac aaaggaacct ttaatagaaa ttggacagca agaaagcgag
     6301 cttctagctt atcctcagtc ctgctcctct gccacaaagt gcacgcagtt gccggccggg
     6361 tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat ggccggcccg
     6421 gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag ctcgtccagg
     6481 ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg gaccgcgctg
     6541 atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac gaagtcccgg
     6601 gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg cgcggtgagc
     6661 accggaacgg cactggtcaa cttggccatg atggctcctc ctgtcaggag aggaaagaga
     6721 agaaggttag tacaattgct atagtgagtt gtattatact atgcagatat actatgccaa
     6781 tgattaattg tcaaactagg gctgcagggt tcatagtgcc acttttcctg cactgcccca
     6841 tctcctgccc accctttccc aggcatagac agtcagtgac ttaccaaact cacaggaggg
     6901 agaaggcaga agcttgagac agacccgcgg gaccgccgaa ctgcgagggg acgtggctag
     6961 ggcggcttct tttatggtgc gccggccctc ggaggcaggg cgctcgggga ggcctagcgg
     7021 ccaatctgcg gtggcaggag gcggggccga aggccgtgcc tgaccaatcc ggagcacata
     7081 ggagtctcag ccccccgccc caaagcaagg ggaagtcacg cgcctgtagc gccagcgtgt
     7141 tgtgaaatgg gggcttgggg gggttggggc cctgattagt caaaacaaac tcccattgac
     7201 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     7261 gtactgccaa aaccgcatca tcatggtaat agcgatgact aatacgtaga tgtactgcca
     7321 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     7381 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     7441 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     7501 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     7561 gccatttacc gtaagttatg taacgcctgc aggttaatta agaacatgtg agcaaaaggc
     7621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     8041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     8101 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     8161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     8221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     8281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatggc tagttaatta
     8341 acatttaaat cagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     8401 ttttgtgtga atcgtaacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
     8461 t
//
