LOCUS       sequence-292627-        7891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgG1) high 
            affinity 5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7891)
  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 7891)
  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-sACE2v5-Fc(IgG1)
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        508..711
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     662..682
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     756..775
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        756..774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3375..3392)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3381..3605
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3651..4079
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3738..3757)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3948..3969
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4088..4108)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4093..4422
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4273..4408
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4335..4354
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4489..5283
                     /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(4543..4562)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5153..5172
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5457..5578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5494..5513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5548..5567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5627..5643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5627..5643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5640..5662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5651..5667
                     /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(5675..5681,5682..5699,5700..5705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5720..5741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5858..5875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6029..6617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6109..6128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6788..7579,7580..7648))
                     /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     7411..7430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7649..7753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7828..7847)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
       61 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      121 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      181 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      301 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      361 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      421 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      481 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      541 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      601 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      661 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      721 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      781 caagctggct agcgccacca tgtcaagctc ttcctggctc cttctcagcc ttgttgctgt
      841 aactgctgct cagtccacca ttgaggaaca ggtcaagaca tttttggaca agtttaacca
      901 cgaagccgaa gacctgttct atcaaagttc acttgcttct tggaattata acaccaatat
      961 tactgaagag aatgtccaaa acatgaataa tgctggggac aaatggtctg cctttttaaa
     1021 ggaacagtcc acacttgccc aaatgtatcc actacaagaa attcagaatc tccaagtcaa
     1081 gcttcagctg caggctcttc agcaaaatgg gtcttcagtg ctctcagaag acaagagcaa
     1141 acggttgaac acaattctaa atacaatgag caccatctac agtactggaa aagtttgtaa
     1201 cccagataat ccacaagaat gcttattact tgaaccaggt ttgaatgaaa taatggcaaa
     1261 cagtttagac tacaatgaga ggctctgggc ttgggaaagc tggagatctg aggtcggcaa
     1321 gcagctgagg ccattatatg aagagtatgt ggtcttgaaa aatgagatgg caagagcaaa
     1381 tcattatgag gactatgggg attattggag aggagactat gaagtaaatg gggtagatgg
     1441 ctatgactac agccgcggcc agttgattga agatgtggaa catacctttg aagagattaa
     1501 accattatat gaacatcttc atgcctatgt gagggcaaag ttgatgaatg cctatccttc
     1561 ctatatcagt ccaattggat gcctccctgc tcatttgctt ggtgatatgt ggggtagatt
     1621 ttggacaaat ctgtactctt tgacagttcc ctttggacag aaaccaaaca tagatgttac
     1681 tgatgcaatg gtggaccagg cctgggatgc acagagaata ttcaaggagg ccgagaagtt
     1741 ctttgtatct gttggtcttc ctaatatgac tcaaggattc tgggaaaatt ccatgctaac
     1801 ggacccagga aatgttcaga aagcagtctg ccatcccaca gcttgggacc tggggaaggg
     1861 cgacttcagg attcttatgt gcacaaaggt gacaatggac gacttcctga cagctcatca
     1921 tgagatgggg catatccagt atgatatggc atatcttgca caaccttttc tgctaagaaa
     1981 tggagctaat gaaggattcc atgaagctgt tggggaaatc atgtcacttt ctgcagccac
     2041 acctaagcat ttaaaatcca ttggtcttct gtcacccgat tttcaagaag acaatgaaac
     2101 agaaataaac ttcctgctca aacaagcact cacgattgtt gggactctgc catttactta
     2161 catgttagag aagtggaggt ggatggtctt taaaggggaa attcccaaag accagtggat
     2221 gaaaaagtgg tgggagatga agcgagagat agttggggtg gtggaacctg tgccccatga
     2281 tgaaacatac tgtgaccccg catctctgtt ccatgtttct aatgattact cattcattcg
     2341 atattacaca aggacccttt accaattcca gtttcaagaa gcactttgtc aagcagctaa
     2401 acatgaaggc cctctgcaca aatgtgacat ctcaaactct acagaagctg gacagaaact
     2461 gttcaatatg ctgaggcttg gaaaatcaga accctggacc ctagcattgg aaaatgttgt
     2521 aggagcaaag aacatgaatg taaggccact gctcaactac tttgagccct tatttacctg
     2581 gctgaaagac cagaacaaga attcttttgt gggatggagt accgactgga gtccatatgc
     2641 agacggatcc gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg
     2701 accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc
     2761 tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg
     2821 gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa
     2881 cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa
     2941 ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc
     3001 caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggagga
     3061 gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat
     3121 cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt
     3181 gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg
     3241 gcagcagggg aacgtcttct catgctccgt gatgcacgag gctctgcaca accactacac
     3301 gcagaagagc ctctccctgt ctccgggtaa atgataactc gagtctagag ggcccgttta
     3361 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3421 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3481 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3541 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3601 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3661 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3721 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3781 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3841 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3901 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3961 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4021 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4081 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4141 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4201 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4261 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4321 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4381 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4441 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4501 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4561 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4621 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4681 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4741 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4801 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4861 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4921 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4981 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5041 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5101 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5161 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5221 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5281 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5341 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5401 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5461 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5521 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5581 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5641 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5701 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5761 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5821 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5881 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5941 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6901 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7081 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7261 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7621 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7681 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7741 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7801 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7861 gtatctgctc cctgcttgtg tgttggaggt c
//
