LOCUS       sequence-292650-        7813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human ACE2 with C9 tag at C-terminal in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7813)
  AUTHORS   Shang J, Ye G, Shi K, Wan Y, Luo C, Aihara H, Geng Q, Auerbach A, Li
            F
  TITLE     Structural basis of receptor recognition by SARS-CoV-2.
  JOURNAL   Nature. 2020 Mar 30. pii: 10.1038/s41586-020-2179-y. doi: 
            10.1038/s41586-020-2179-y.
  PUBMED    32225175
REFERENCE   2  (bases 1 to 7813)
  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"; journalName: "Nature. 
            2020 Mar 30. pii: 10.1038/s41586-020-2179-y. doi: 
            10.1038/s41586-020-2179-y."
COMMENT     Sequence Label: pcDNA3.1-hACE2
FEATURES             Location/Qualifiers
     source          1..7813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        170..549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        550..753
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     704..724
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     798..817
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        798..816
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3341..3358)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3347..3571
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3617..4045
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3704..3723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3914..3935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4054..4074)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4059..4388
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4239..4374
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4301..4320
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4509..4528)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5117..5136
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5421..5542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5458..5477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5512..5531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5591..5607)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5591..5607)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5604..5626)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5615..5631
                     /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(5639..5645,5646..5663,5664..5669))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5684..5705
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5822..5839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5993..6581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6073..6092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6752..7543,7544..7612))
                     /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     7375..7394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7613..7717)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7792..7811)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
       61 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
      121 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      181 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      241 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      301 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      361 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      421 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      481 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      541 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      601 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      661 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      721 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      781 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      841 aacgggccct ctagaccatg tcaagctctt cctggctcct tctcagcctt gttgctgtaa
      901 ctgctgctca gtccaccatt gaggaacagg ccaagacatt tttggacaag tttaaccacg
      961 aagccgaaga cctgttctat caaagttcac ttgcttcttg gaattataac accaatatta
     1021 ctgaagagaa tgtccaaaac atgaataatg ctggggacaa atggtctgcc tttttaaagg
     1081 aacagtccac acttgcccaa atgtatccac tacaagaaat tcagaatctc acagtcaagc
     1141 ttcagctgca ggctcttcag caaaatgggt cttcagtgct ctcagaagac aagagcaaac
     1201 ggttgaacac aattctaaat acaatgagca ccatctacag tactggaaaa gtttgtaacc
     1261 cagataatcc acaagaatgc ttattacttg aaccaggttt gaatgaaata atggcaaaca
     1321 gtttagacta caatgagagg ctctgggctt gggaaagctg gagatctgag gtcggcaagc
     1381 agctgaggcc attatatgaa gagtatgtgg tcttgaaaaa tgagatggca agagcaaatc
     1441 attatgagga ctatggggat tattggagag gagactatga agtaaatggg gtagatggct
     1501 atgactacag ccgcggccag ttgattgaag atgtggaaca tacctttgaa gagattaaac
     1561 cattatatga acatcttcat gcctatgtga gggcaaagtt gatgaatgcc tatccttcct
     1621 atatcagtcc aattggatgc ctccctgctc atttgcttgg tgatatgtgg ggtagatttt
     1681 ggacaaatct gtactctttg acagttccct ttggacagaa accaaacata gatgttactg
     1741 atgcaatggt ggaccaggcc tgggatgcac agagaatatt caaggaggcc gagaagttct
     1801 ttgtatctgt tggtcttcct aatatgactc aaggattctg ggaaaattcc atgctaacgg
     1861 acccaggaaa tgttcagaaa gcagtctgcc atcccacagc ttgggacctg gggaagggcg
     1921 acttcaggat ccttatgtgc acaaaggtga caatggacga cttcctgaca gctcatcatg
     1981 agatggggca tatccagtat gatatggcat atgctgcaca accttttctg ctaagaaatg
     2041 gagctaatga aggattccat gaagctgttg gggaaatcat gtcactttct gcagccacac
     2101 ctaagcattt aaaatccatt ggtcttctgt cacccgattt tcaagaagac aatgaaacag
     2161 aaataaactt cctgctcaaa caagcactca cgattgttgg gactctgcca tttacttaca
     2221 tgttagagaa gtggaggtgg atggtcttta aaggggaaat tcccaaagac cagtggatga
     2281 aaaagtggtg ggagatgaag cgagagatag ttggggtggt ggaacctgtg ccccatgatg
     2341 aaacatactg tgaccccgca tctctgttcc atgtttctaa tgattactca ttcattcgat
     2401 attacacaag gaccctttac caattccagt ttcaagaagc actttgtcaa gcagctaaac
     2461 atgaaggccc tctgcacaaa tgtgacatct caaactctac agaagctgga cagaaactgt
     2521 tcaatatgct gaggcttgga aaatcagaac cctggaccct agcattggaa aatgttgtag
     2581 gagcaaagaa catgaatgta aggccactgc tcaactactt tgagccctta tttacctggc
     2641 tgaaagacca gaacaagaat tcttttgtgg gatggagtac cgactggagt ccatatgcag
     2701 accaaagcat caaagtgagg ataagcctaa aatcagctct tggagataaa gcatatgaat
     2761 ggaacgacaa tgaaatgtac ctgttccgat catctgttgc atatgctatg aggcagtact
     2821 ttttaaaagt aaaaaatcag atgattcttt ttggggagga ggatgtgcga gtggctaatt
     2881 tgaaaccaag aatctccttt aatttctttg tcactgcacc taaaaatgtg tctgatatca
     2941 ttcctagaac tgaagttgaa aaggccatca ggatgtcccg gagccgtatc aatgatgctt
     3001 tccgtctgaa tgacaacagc ctagagtttc tggggataca gccaacactt ggacctccta
     3061 accagccccc tgtttccata tggctgattg tttttggagt tgtgatggga gtgatagtgg
     3121 ttggcattgt catcctgatc ttcactggga tcagagatcg gaagaagaaa aataaagcaa
     3181 gaagtggaga aaatccttat gcctccatcg atattagcaa aggagaaaat aatccaggat
     3241 tccaaaacac tgatgatgtt cagacctcct ttggtaccga gacctcccag gtggcgcccg
     3301 cttaagcttg gatccgcgtt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     3361 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3421 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3481 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3541 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3601 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3661 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3721 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3781 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3841 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3901 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3961 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4021 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4081 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4141 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4261 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4321 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4381 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4441 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4501 tggagaggct attcggctat gactgggcaa cagacaatcg gctgctctga tgccgccgtg
     4561 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4621 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4681 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4741 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4801 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4861 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4921 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4981 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5041 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5101 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5161 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5221 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5281 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5341 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5401 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5521 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5581 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5641 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5701 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5761 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5821 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5881 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7741 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     7801 ctctgatgcc gca
//
