LOCUS       sequence-314771-        6598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ACE2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6598)
  AUTHORS   Yurkovetskiy L, Wang X, Pascal KE, Tomkins-Tinch C, Nyalile TP, Wang
            Y, Baum A, Diehl WE, Dauphin A, Carbone C, Veinotte K, Egri SB, 
            Schaffner SF, Lemieux JE, Munro JB, Rafique A, Barve A, Sabeti PC, 
            Kyratsous CA, Dudkina NV, Shen K, Luban J
  TITLE     Structural and Functional Analysis of the D614G SARS-CoV-2 Spike 
            Protein Variant.
  JOURNAL   Cell. 2020 Sep 15. pii: S0092-8674(20)31229-0. doi: 
            10.1016/j.cell.2020.09.032.
  PUBMED    32991842
REFERENCE   2  (bases 1 to 6598)
  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: "Cell. 2020 
            Sep 15. pii: S0092-8674(20)31229-0. doi: 
            10.1016/j.cell.2020.09.032."
COMMENT     Sequence Label: pCAGGS-ACE2-his expression plasmid
FEATURES             Location/Qualifiers
     source          1..6598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3589..3606
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3619..3638)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3684..3739
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3738..3757)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4100..4116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4100..4116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4113..4135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4124..4140
                     /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(4148..4154,4155..4172,4173..4178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4193..4214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4273..4468
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4319..4454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4474..4608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4511..4530)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4565..4584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4676..4693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4847..5435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4927..4946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5606..6397,6398..6466))
                     /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     6229..6248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6467..6571)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cgccaccatg
     1741 agtagctcaa gctggcttct gctttccttg gttgccgtca cggccgccca atcaaccata
     1801 gaagagcaag caaagacatt tttggacaag tttaaccacg aagccgagga tcttttttat
     1861 cagtcttctt tggcatcttg gaactacaac acgaacatca cggaagaaaa cgtgcaaaac
     1921 atgaataatg ccggtgataa gtggtcagcc ttcctgaaag agcagtccac tctggcccaa
     1981 atgtaccctc ttcaggagat ccagaatctc actgttaagt tgcaattgca agccttgcag
     2041 cagaatggga gtagcgtatt gagtgaggat aagagcaaga gactgaatac cattctgaat
     2101 acaatgagca caatttatag caccggcaaa gtctgcaatc cggacaaccc gcaggaatgc
     2161 ttgctgttgg agcccggatt gaatgaaatc atggctaaca gcttggatta taacgagagg
     2221 ctttgggcat gggagtcatg gagaagtgag gtagggaaac aacttagacc cctttacgag
     2281 gagtacgttg tactcaaaaa cgaaatggca agggcaaacc attacgagga ttacggtgat
     2341 tattggagag gagactatga agtcaacggg gtcgatggtt acgattattc acgcggacag
     2401 ctcatcgaag acgtagagca tacgtttgaa gaaatcaagc cactctatga acacttgcac
     2461 gcctacgttc gcgctaagtt gatgaatgct tatccgtcat atatctcacc aatcggatgc
     2521 ctcccggcac acctgctggg ggacatgtgg ggtcggttct ggaccaacct ctacagtttg
     2581 acagtacctt tcggtcaaaa accaaatatt gacgtaacgg acgcgatggt cgaccaagcc
     2641 tgggatgcgc agcggatctt taaagaagcg gagaaattct ttgtatcagt cgggttgcct
     2701 aatatgactc agggcttctg ggaaaactct atgctcacag atccgggtaa tgtacaaaaa
     2761 gccgtatgcc atcccactgc atgggacctc ggcaaaggag acttccggat actcatgtgc
     2821 actaaagtca cgatggatga cttcctgacg gcacatcatg agatggggca catacagtat
     2881 gacatggcat atgcagccca accgtttctc cttcgcaacg gcgcgaacga aggctttcac
     2941 gaggctgtag gggaaattat gtctctgtct gctgccacgc cgaagcatct caaatccata
     3001 ggtctccttt caccggattt ccaagaggat aacgagaccg agatcaactt tcttctgaag
     3061 caggcattga cgatagtagg gactctgccg ttcacgtata tgctcgaaaa atggcggtgg
     3121 atggtattca agggagagat accgaaagac cagtggatga agaaatggtg ggaaatgaag
     3181 agagaaatag tcggtgtcgt cgagccagtc cctcatgacg agacgtactg cgacccagca
     3241 agcctttttc atgtttcaaa cgactacagt ttcatacggt attatactag aacgctctat
     3301 caattccagt tccaagaagc cctgtgccag gccgctaagc acgaaggacc acttcataag
     3361 tgcgatatca gtaactcaac cgaagcaggc cagaagcttt ttaatatgtt gcgcctcggc
     3421 aaaagcgagc cgtggacgtt ggctttggag aacgtcgtcg gagcgaagaa catgaacgta
     3481 cggcctctcc tgaactattt cgaacctctc tttacgtggt tgaaagacca aaacaagaac
     3541 tcctttgtgg ggtggtcaac cgattggtca ccttacgctg atggaagcca tcatcaccat
     3601 catcattgag ctagcagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     3661 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     3721 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     3781 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     3841 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     3901 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     3961 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     4021 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     4081 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4141 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4201 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4261 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4321 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4381 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4441 cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt
     4501 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4561 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
     4621 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4681 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4741 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4801 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4861 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4921 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4981 ccctgccact taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5041 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5101 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5161 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5221 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5281 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5341 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5401 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5461 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5521 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5581 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5641 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5701 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5761 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5821 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5881 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5941 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6001 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6061 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6121 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6181 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6241 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6301 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6361 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6421 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6481 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6541 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgg
//
