LOCUS       sequence-333795-        9005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The extracellular domain of Angiotensin converting enzyme 2 (ACE2) 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9005)
  AUTHORS   Kolesov DE, Sinegubova MV, Dayanova LK, Dolzhikova IV, Vorobiev II, 
            Orlova NA
  TITLE     Fast and Accurate Surrogate Virus Neutralization Test Based on 
            Antibody-Mediated Blocking of the Interaction of ACE2 and SARS-CoV-2
            Spike Protein RBD.
  JOURNAL   Diagnostics (Basel). 2022 Feb 3;12(2). pii: diagnostics12020393. 
            doi: 10.3390/diagnostics12020393.
  PUBMED    35204485
REFERENCE   2  (bases 1 to 9005)
  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: "Diagnostics
            (Basel)."; date: "2022-02-3"; volume: "12(2). pii: 
            diagnostics12020393. doi"; pages: " 10.3390/diagnostics12020393"
COMMENT     Sequence Label: pTF-ACE2s
FEATURES             Location/Qualifiers
     source          1..9005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2265..2288
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2298..2327
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    2355..2928
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2378..2930
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2522..2539)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2749..2768
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2945..2954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2951..3514
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     primer_bind     complement(4183..4200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4354..4942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4434..4453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5113..5904,5905..5973))
                     /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     5736..5755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5974..6078)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttttttaag gtgttgtgaa aacctcgagg ccgccaccat gtcaagctct tcctggctcc
       61 ttctcagcct tgttgctgta actgctgctc agtccaccat tgaggaacag gccaagacat
      121 ttttggacaa gtttaaccac gaagccgaag acctgttcta tcaaagttca cttgcttctt
      181 ggaattataa caccaatatt actgaagaga atgtccaaaa catgaataat gctggggaca
      241 aatggtctgc ctttttaaag gaacagtcca cacttgccca aatgtatcca ctacaagaaa
      301 ttcagaatct cacagtcaag cttcagctgc aggctcttca gcaaaatggg tcttcagtgc
      361 tctcagaaga caagagcaaa cggttgaaca caattctaaa tacaatgagc accatctaca
      421 gtactggaaa agtttgtaac ccagataatc cacaagaatg cttattactt gaaccaggtt
      481 tgaatgaaat aatggcaaac agtttagact acaatgagag gctctgggct tgggaaagct
      541 ggagatctga ggtcggcaag cagctgaggc cattatatga agagtatgtg gtcttgaaaa
      601 atgagatggc aagagcaaat cattatgagg actatgggga ttattggaga ggagactatg
      661 aagtaaatgg ggtagatggc tatgactaca gccgcggcca gttgattgaa gatgtggaac
      721 atacctttga agagattaaa ccattatatg aacatcttca tgcctatgtg agggcaaagt
      781 tgatgaatgc ctatccttcc tatatcagtc caattggatg cctccctgct catttgcttg
      841 gtgatatgtg gggtagattt tggacaaatc tgtactcttt gacagttccc tttggacaga
      901 aaccaaacat agatgttact gatgcaatgg tggaccaggc ctgggatgca cagagaatat
      961 tcaaggaggc cgagaagttc tttgtatctg ttggtcttcc taatatgact caaggattct
     1021 gggaaaattc catgctaacg gacccaggaa atgttcagaa agcagtctgc catcccacag
     1081 cttgggacct ggggaagggc gacttcagga tccttatgtg cacaaaggtg acaatggacg
     1141 acttcctgac agctcatcat gagatggggc atatccagta tgatatggca tatgctgcac
     1201 aaccttttct gctaagaaat ggagctaatg aaggattcca tgaagctgtt ggggaaatca
     1261 tgtcactttc tgcagccaca cctaagcatt taaaatccat tggtcttctg tcacccgatt
     1321 ttcaagaaga caatgaaaca gaaataaact tcctgctcaa acaagcactc acgattgttg
     1381 ggactctgcc atttacttac atgttagaga agtggaggtg gatggtcttt aaaggggaaa
     1441 ttcccaaaga ccagtggatg aaaaagtggt gggagatgaa gcgagagata gttggggtgg
     1501 tggaacctgt gccccatgat gaaacatact gtgaccccgc atctctgttc catgtttcta
     1561 atgattactc attcattcga tattacacaa ggacccttta ccaattccag tttcaagaag
     1621 cactttgtca agcagctaaa catgaaggcc ctctgcacaa atgtgacatc tcaaactcta
     1681 cagaagctgg acagaaactg ttcaatatgc tgaggcttgg aaaatcagaa ccctggaccc
     1741 tagcattgga aaatgttgta ggagcaaaga acatgaatgt aaggccactg ctcaactact
     1801 ttgagccctt atttacctgg ctgaaagacc agaacaagaa ttcttttgtg ggatggagta
     1861 ccgactggag tccatatgca gaccaaagca tcaaagtgag gataagccta aaatcagctc
     1921 ttggagataa agcatatgaa tggaacgaca atgaaatgta cctgttccga tcatctgttg
     1981 catatgctat gaggcagtac tttttaaaag taaaaaatca gatgattctt tttggggagg
     2041 aggatgtgcg agtggctaat ttgaaaccaa gaatctcctt taatttcttt gtcactgcac
     2101 ctaaaaatgt gtctgatatc attcctagaa ctgaagttga aaaggccatc aggatgtccc
     2161 ggagccgtat caatgatgct ttccgtctga atgacaacag cctagagttt ctggggatac
     2221 agccaacact tggacctcct aaccagcccc ctgtttccgc tagcgattac aaggatgacg
     2281 acgataaggg aggatcccat caccatcacc atcaccatca tcaccactaa ctagcggccg
     2341 cgcagttaac gccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg
     2401 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     2461 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     2521 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     2581 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     2641 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     2701 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     2761 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     2821 ctggggcctc ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     2881 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaag
     2941 atctgccacc atggttcgac cattgaactg catcgtcgcc gtgtcccaaa atatggggat
     3001 tggcaagaac ggagacctac cctggcctcc gctcaggaac gagttcaagt acttccaaag
     3061 aatgaccaca acctcttcag tggaaggtaa acagaatctg gtgattatgg gtaggaaaac
     3121 ctggttctcc attcctgaga agaatcgacc tctaaaggac agaattaata tagttctcag
     3181 tagagaactc aaagaaccac cacgaggagc tcattttctt gccaaaagtt tggatgatgc
     3241 cttaagactt attgaacaac cggaattggc aagtaaagta gacatggttt ggatagtcgg
     3301 aggcagttct gtttaccagg aagccatgaa tcaaccaggc cacctcagac tctttgtgac
     3361 aaggatcatg caggaatttg aaagtgacac gtttttccca gaaattgatt tggggaaata
     3421 taaacttctc ccagaatacc caggcgtcct ctctgaggtc caggaggaaa aaggcatcaa
     3481 gtataagttt gaagtctacg agaagaaaga ctaatctagc atattacccc taacacctgc
     3541 caccccagtc ttaatcagtg gtggaagaac ggtctcagaa ctgtttgtct caattggcca
     3601 tttaagttta atagtgaaag actggttaat gataacaatg catcggaaaa ccttcaggag
     3661 gaaaggagaa tgttttgtgg aacatttttg tgtgtgtggc agttttaagt tattagtttt
     3721 caaaatcagt actttttaat ggaaacaact tgaccaaaaa tctgtcacag aattttgaga
     3781 cccattaaaa tacaagttta atgagaagtc tgtctctgtt aatgctgaag tcattactaa
     3841 gtgcttagct tagcaaggta tgtggatgcc catttgtgtt ccaagggatt ggactgttca
     3901 tcaggaccca gagctgagtt tcaagggctc aagagatggc ttattacctg tgggtgtctt
     3961 gaaggttctg gttgggacaa attaggaatg tttttggcag acatggtgac tacccttcat
     4021 ctgggtgagt tcagttgatt tgtcttgagc ctttggggtt tacacaagta aatgacatca
     4081 tacagttagt gtattgttag tgaatattaa tatatgaggc aggctttgct ctagacatat
     4141 gtcgacgtcc ggagctctgg cgcgccatgg ctcttccgct tcctcgctca ctgactcgct
     4201 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4261 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4321 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4381 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4441 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4501 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4561 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4621 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4681 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4741 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4801 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4861 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4921 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4981 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5041 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5101 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5161 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5221 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5281 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5341 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5401 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5461 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5521 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5581 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5641 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5701 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5761 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5821 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5881 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5941 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6001 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6061 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgcatctc atcacggtca
     6121 cgcatggctg cgcccgcgca gcgccccccg accccccccc gatccccgga acgccagccg
     6181 ccatccaccg cccgcagccc ccggagcccg cggaccccga ccccccgccg ctcgcccgcc
     6241 gttgcgccgc tctctgcggg ggggctaggg ggccgcgggg ggaaggccac gccccctcca
     6301 ctttttccag gaatgcgcgg catgacccgc cccccccatg acccgccaac acccccccga
     6361 tgccccccgg gggtctttcc tggggggcct ttgtcagggt tgcctgtgtc acccacgggc
     6421 cccttggcca cgcttgcctg ggtcaccccg gagccccggg tcacgtttgc cggggtcagc
     6481 cctggggcgc ttcttcacgc tccattgcca tgcagaattc ccacatgtgg acaccggaca
     6541 gtaggtcctc aaatgctcct tattaggttg gttcaataat atcaattgtt tgttactagg
     6601 cagtgatgtt gtacatctgg aggagatctc ttgagcccat aatcaggtta ttaggaataa
     6661 atactctaag gctaaaaatg tagcttagtg ataagagtgc ttgcctggtg tgctgagacc
     6721 ctcggttcca tctccacaac cccatattcc attacaaaat accttttcac cgtccctagc
     6781 attaagaaac aaaacaacaa agaagttttt ctttcttctg agatcctgcc cggagaggca
     6841 tttaaaactg gccagggcca aaaaaaaaaa aaaaagaaaa aaaagaaaag aaaacaggct
     6901 agggccggca tggtggcgca cgcctttaat cccagcacgc aggaggcaga ggcagggcgg
     6961 atctctgtga gtttgaggtc agcctggtct acctagtgag tttcagggca cccagggcta
     7021 aagagactgt ctcaaaaaca aaacagccac acaatcagaa ccacagcaaa acgcagttat
     7081 gatccttgga actgtaggaa tgacaagcat ttaaataata ggacgagcca tttttgagaa
     7141 gctctgattt cacaagtgtc agggatgggc tctgggcgag taagattgct aatgctggcc
     7201 tctaaatgag accacgtgga gttgattaga ttcttttcat gttcctcgtg ctctatcaaa
     7261 taactgtacc caaatacaca cacacacaca cacacacaat gcgcgcacac acaaaatcct
     7321 tttttagctt aagaagccca gaatcagaag taaagctaac tgtgggactt aagtattatt
     7381 ctgaacggaa ctcccagggc gtgaagcgcg cttcaggctt ccagagaagc agctggcgct
     7441 ggatggaatg aaccaagagg ccagcacagg ggcagatccg tcgagctctc ggccaccgag
     7501 ctgagccctt aggttctggg gctgggaagg gtccctagga ttgtgcacct ctcccgcggg
     7561 ggacaagcag ggggatggcg gggctgacgt cgggaggtgg cctccacggg aagggacacc
     7621 cggatctcga cacagccttg gcagtggagt caggaagggt agggacagat tctggacgcc
     7681 ctcttggcca gctcctcacc gccccacccc cgactggagc cgagagtaat tcatacaaaa
     7741 ggagggatcg ccttcgcccc tgggaatccc agggaccgtc gctaaattct ggccggcctc
     7801 ccagcccgga accgctgtgc ccgcccagcg cggcgggagg agcctgcgcc tagggcggat
     7861 cgcgggtcgg cgggagagca caagcccaca gtccccggcg gtgggggagg ggcgcgctga
     7921 gcgggggccc gggagccagc gcggggcaaa ctgggaaagt ggcgtcgtgt gctggctccg
     7981 ccctcttccc gagggtgggg gagaacggta taaaagtgcg gtagtcgcgt tggacgttct
     8041 ttttcgcaac gggtttgccg tcagaacgca ggtgagtggc gggtgtggcc tccgcgggcc
     8101 cgggctccct cctttgagcg gggtcgggcc gccgtgcggg tgtcgtcggc cgggcttctc
     8161 tgcgagcgtt cccgccctgg atggcgggct gtgcgggagg gcgagggggg gaggcctggc
     8221 ggcggccccg gagcctcgcc tcgtgtcggg cgtgaggcct agcgtggctt ccgccccgcc
     8281 gcgtgccacc gcggccgcgc tttgctgtct gcccggctgc cctcgattgc ctgcccgcgg
     8341 cccgggccaa caaagggagg gcgtggagct ggctggtagg gagccccgta gtccgcatgt
     8401 cgggcaggga gagcggcagc agtcgggggg gggaccgggc ccgcccgtcc cgcagcacat
     8461 gtccaacgcc gcctggacgg gtagcggcct gtgtcctgat aaggcggccg ggcggtgggt
     8521 tttagatgcc gggttcaggt ggccccgggt cccggcccgg tctggccagt accccgtagt
     8581 ggcttagctc cgaggagggc gaggcccgcc cgcccggcac cagttgcgtg cgcggaaaga
     8641 tggccgctcc cgggccctgt agcaaggagc tcaaaatgga ggacgcggca gcccggcgga
     8701 gcggggcggg tgagtcaccc acacaaagga agagggcctt gcccctcgcc ggccgctgct
     8761 tcctgtgacc ccgtggtgta ccggccgcac ttcagtcacc ccgggcgctc tttcggagca
     8821 ccgctggcct ccgctggggg aggggatctg tctaatggcg ttggagtttg ctcacatttg
     8881 gtgggtggag actgtagcca ggccagcctg gccatgaaag taattcttgg aatttgccca
     8941 ttttgagttt ggagcgaagc tgattgacaa agctgcttag ccgttcaaag gtattcttcg
     9001 aactt
//
