LOCUS       sequence-292584-        7210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2 (protease domain) high
            affinity variant 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7210)
  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 7210)
  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-sACE2v1
FEATURES             Location/Qualifiers
     source          1..7210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..479
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        480..683
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     634..654
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     728..747
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        728..746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2666..2683)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2672..2896
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2942..3370
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3029..3048)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3239..3260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3379..3399)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3384..3713
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3564..3699
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3626..3645
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3780..4574
                     /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(3834..3853)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4444..4463
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4748..4869
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4785..4804)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4839..4858
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4918..4934)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4918..4934)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4931..4953)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4942..4958
                     /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(4966..4972,4973..4990,4991..4996))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5011..5032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5149..5166)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5320..5908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5400..5419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6079..6870,6871..6939))
                     /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     6702..6721
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6940..7044)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7119..7138)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
       61 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      121 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      181 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      241 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      301 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      361 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      421 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      481 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      541 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      601 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      661 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      721 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgccac catgtcaagc
      781 tcttcctggc tccttctcag ccttgttgct gtaactgctg ctcagtccac cattgaggaa
      841 caggccaaga catttttgga caagtttaac gccgaagccg aagacctgtt ctatcaaagt
      901 tcacttgctt cttggaatta taacaccaat attactgaag agaatgtcca aaacatgaat
      961 aatgctgggg acaaatggtc tgccttttta aaggaacagt ccacacttgc ccaaatgtat
     1021 ccactacaag aaattcagaa tctccaagtc aagcttcagc tgcaggctct tcagcaaaat
     1081 gggtcttcag tgctctcaga agacaagagc aaacggttga acacaattct aaatacaatg
     1141 agcaccatct acagtactgg aaaagtttgt aacccagata atccacaaga atgcttatta
     1201 cttgaaccag gtttgaatga aataatggca aacagtttag actacaatga gaggctctgg
     1261 gcttgggaaa gctggagatc tgaggtcggc aagcagctga ggccattata tgaagagtat
     1321 gtggtcttga aaaatgagat ggcaagagca aatcattatg aggactatgg ggattattgg
     1381 agaggagact atgaagtaaa tggggtagat ggctatgact acagccgcgg ccagttgatt
     1441 gaagatgtgg aacatacctt tgaagagatt aaaccattat atgaacatct tcatgcctat
     1501 gtgagggcaa agttgatgaa tgcctatcct tcctatatca gtccaattgg atgcctccct
     1561 gctcatttgc ttggtgatat gtggggtaga ttttggacaa atctgtactc tttgacagtt
     1621 ccctttggac agaaaccaaa catagatgtt actgatgcaa tggtggacca ggcctgggat
     1681 gcacagagaa tattcaagga ggccgagaag ttctttgtat ctgttggtct tcctaatatg
     1741 actccaggat tctgggaaaa ttccatgcta acggacccag gaaatgttca gaaagcagtc
     1801 tgccatccca cagcttggga cctggggaag ggcgacttca ggattcttat gtgcacaaag
     1861 gtgacaatgg acgacttcct gacagctcat catgagatgg ggcatatcca gtatgatatg
     1921 gcatatcttg cacaaccttt tctgctaaga aatggagcta atgaaggatt ccatgaagct
     1981 gttggggaaa tcatgtcact ttctgcagcc acacctaagc atttaaaatc cattggtctt
     2041 ctgtcacccg attttcaaga agacaatgaa acagaaataa acttcctgct caaacaagca
     2101 ctcacgattg ttgggactct gccatttact tacatgttag agaagtggag gtggatggtc
     2161 tttaaagggg aaattcccaa agaccagtgg atgaaaaagt ggtgggagat gaagcgagag
     2221 atagttgggg tggtggaacc tgtgccccat gatgaaacat actgtgaccc cgcatctctg
     2281 ttccatgttt ctaatgatta ctcattcatt cgatattaca caaggaccct ttaccaattc
     2341 cagtttcaag aagcactttg tcaagcagct aaacatgaag gccctctgca caaatgtgac
     2401 atctcaaact ctacagaagc tggacagaaa ctgttcaata tgctgaggct tggaaaatca
     2461 gaaccctgga ccctagcatt ggaaaatgtt gtaggagcaa agaacatgaa tgtaaggcca
     2521 ctgctcaact actttgagcc cttatttacc tggctgaaag accagaacaa gaattctttt
     2581 gtgggatgga gtaccgactg gagtccatat gcagacggat cctgataact cgagtctaga
     2641 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2701 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2761 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2821 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2881 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2941 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3001 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3061 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3121 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3181 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3241 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3301 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3361 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3421 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3541 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3601 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3661 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3721 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3781 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3841 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3901 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3961 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4021 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4081 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4141 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4201 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4261 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4321 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4381 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4441 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4501 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4561 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4621 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4681 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4741 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4801 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4861 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4921 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4981 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5041 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5101 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5161 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5221 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6061 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6121 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6181 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6241 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6301 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6361 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6421 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6481 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6541 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6601 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6661 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6721 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6781 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6841 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6901 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6961 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7021 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7081 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     7141 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7201 aaaatttaag
//
