LOCUS       sequence-292583-        7210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2 (protease domain).
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-sACE2(WT)
FEATURES             Location/Qualifiers
     source          1..7210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     780..799
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        780..798
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2718..2735)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2724..2948
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2994..3422
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3081..3100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3291..3312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3431..3451)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3436..3765
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3616..3751
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3678..3697
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3832..4626
                     /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(3886..3905)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4496..4515
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4800..4921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4837..4856)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4891..4910
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4970..4986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4970..4986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4983..5005)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4994..5010
                     /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(5018..5024,5025..5042,5043..5048))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5063..5084
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5201..5218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5372..5960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5452..5471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6131..6922,6923..6991))
                     /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     6754..6773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6992..7096)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7171..7190)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
       61 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
      121 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
      781 aatacgactc actataggga gacccaagct ggctagcgcc accatgtcaa gctcttcctg
      841 gctccttctc agccttgttg ctgtaactgc tgctcagtcc accattgagg aacaggccaa
      901 gacatttttg gacaagttta accacgaagc cgaagacctg ttctatcaaa gttcacttgc
      961 ttcttggaat tataacacca atattactga agagaatgtc caaaacatga ataatgctgg
     1021 ggacaaatgg tctgcctttt taaaggaaca gtccacactt gcccaaatgt atccactaca
     1081 agaaattcag aatctcacag tcaagcttca gctgcaggct cttcagcaaa atgggtcttc
     1141 agtgctctca gaagacaaga gcaaacggtt gaacacaatt ctaaatacaa tgagcaccat
     1201 ctacagtact ggaaaagttt gtaacccaga taatccacaa gaatgcttat tacttgaacc
     1261 aggtttgaat gaaataatgg caaacagttt agactacaat gagaggctct gggcttggga
     1321 aagctggaga tctgaggtcg gcaagcagct gaggccatta tatgaagagt atgtggtctt
     1381 gaaaaatgag atggcaagag caaatcatta tgaggactat ggggattatt ggagaggaga
     1441 ctatgaagta aatggggtag atggctatga ctacagccgc ggccagttga ttgaagatgt
     1501 ggaacatacc tttgaagaga ttaaaccatt atatgaacat cttcatgcct atgtgagggc
     1561 aaagttgatg aatgcctatc cttcctatat cagtccaatt ggatgcctcc ctgctcattt
     1621 gcttggtgat atgtggggta gattttggac aaatctgtac tctttgacag ttccctttgg
     1681 acagaaacca aacatagatg ttactgatgc aatggtggac caggcctggg atgcacagag
     1741 aatattcaag gaggccgaga agttctttgt atctgttggt cttcctaata tgactcaagg
     1801 attctgggaa aattccatgc taacggaccc aggaaatgtt cagaaagcag tctgccatcc
     1861 cacagcttgg gacctgggga agggcgactt caggattctt atgtgcacaa aggtgacaat
     1921 ggacgacttc ctgacagctc atcatgagat ggggcatatc cagtatgata tggcatatgc
     1981 tgcacaacct tttctgctaa gaaatggagc taatgaagga ttccatgaag ctgttgggga
     2041 aatcatgtca ctttctgcag ccacacctaa gcatttaaaa tccattggtc ttctgtcacc
     2101 cgattttcaa gaagacaatg aaacagaaat aaacttcctg ctcaaacaag cactcacgat
     2161 tgttgggact ctgccattta cttacatgtt agagaagtgg aggtggatgg tctttaaagg
     2221 ggaaattccc aaagaccagt ggatgaaaaa gtggtgggag atgaagcgag agatagttgg
     2281 ggtggtggaa cctgtgcccc atgatgaaac atactgtgac cccgcatctc tgttccatgt
     2341 ttctaatgat tactcattca ttcgatatta cacaaggacc ctttaccaat tccagtttca
     2401 agaagcactt tgtcaagcag ctaaacatga aggccctctg cacaaatgtg acatctcaaa
     2461 ctctacagaa gctggacaga aactgttcaa tatgctgagg cttggaaaat cagaaccctg
     2521 gaccctagca ttggaaaatg ttgtaggagc aaagaacatg aatgtaaggc cactgctcaa
     2581 ctactttgag cccttattta cctggctgaa agaccagaac aagaattctt ttgtgggatg
     2641 gagtaccgac tggagtccat atgcagacgg atcctgataa ctcgagtcta gagggcccgt
     2701 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2761 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2821 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2881 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2941 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3001 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3061 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3121 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3181 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3241 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3301 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3361 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3421 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3481 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3541 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3601 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3661 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3721 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3781 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3841 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3901 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3961 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4021 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4081 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4141 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4201 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4261 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4321 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4381 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4441 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4501 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4561 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4621 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4681 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4741 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4801 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4861 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4921 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4981 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5041 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5101 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5161 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5221 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5281 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5341 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5401 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5461 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5521 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5581 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5641 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5701 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5761 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5821 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5881 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5941 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6001 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6061 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6121 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6181 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6241 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6301 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6361 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6421 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6481 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6541 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6601 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6661 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6721 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6781 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6841 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6901 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6961 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7021 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7081 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7141 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7201 ccagtatctg
//
