LOCUS       sequence-458394-        8180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human Ace2 protein under CMV promoter fused 
            with IgG1, tagged with 6His, and released extracellular environment 
            after expression for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8180)
  AUTHORS   Ozcelik CE, Araz CZ, Yilmaz O, Gulyuz S, Ozdamar P, Salmanli E, 
            Ozkul A, Seker UOS
  TITLE     Screening Peptide Drug Candidates To Neutralize Whole Viral Agents: 
            A Case Study with Severe Acute Respiratory Syndrome Coronavirus 2 
            (SARS-CoV-2).
  JOURNAL   ACS Pharmacol Transl Sci. 2024 Mar 21;7(4):1032-1042. doi: 
            10.1021/acsptsci.3c00317. eCollection 2024 Apr 12.
  PUBMED    38633598
REFERENCE   2  (bases 1 to 8180)
  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: 
            "10.1021/acsptsci.3c00317"; journalName: "ACS Pharmacol Transl Sci";
            date: "2024-03-21- 21"; volume: "7"; issue: "4"; pages: "1032-1042"
COMMENT     Sequence Label: AP2u2-CMV-Ace2-IgG1
FEATURES             Location/Qualifiers
     source          1..8180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     645..664
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        645..663
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2938..2961
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     intron          3738..3967
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3950..3969
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(4040..4614,4615..4617,4618..4626)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4207..4224)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4434..4453
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4649..5452
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4712..4731)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5322..5341
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5582..5703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5619..5638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5805..5821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5805..5821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5818..5840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5829..5845
                     /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(5853..5859,5860..5877,5878..5883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5898..5919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6036..6053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6207..6795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6287..6306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6966..7757,7758..7826))
                     /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     7589..7608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7827..7931)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8006..8025)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      661 agggagaccc aagcttggta ccgagctcgg atcgatatcg ccgccaccat gtcaagctct
      721 tcctggctcc ttctcagcct tgttgctgta actgctgctc agtccaccat tgaggaacag
      781 gccaagacat ttttggacaa gtttaaccac gaagccgaag acctgttcta tcaaagttca
      841 cttgcttctt ggaattataa caccaatatt actgaagaga atgtccaaaa catgaataat
      901 gctggggaca aatggtctgc ctttttaaag gaacagtcca cacttgccca aatgtatcca
      961 ctacaagaaa ttcagaatct cacagtcaag cttcagctgc aggctcttca gcaaaatggg
     1021 tcttcagtgc tctcagaaga caagagcaaa cggttgaaca caattctaaa tacaatgagc
     1081 accatctaca gtactggaaa agtttgtaac ccagataatc cacaagaatg cttattactt
     1141 gaaccaggtt tgaatgaaat aatggcaaac agtttagact acaatgagag gctctgggct
     1201 tgggaaagct ggagatctga ggtcggcaag cagctgaggc cattatatga agagtatgtg
     1261 gtcttgaaaa atgagatggc aagagcaaat cattatgagg actatgggga ttattggaga
     1321 ggagactatg aagtaaatgg ggtagatggc tatgactaca gccgcggcca gttgattgaa
     1381 gatgtggaac atacctttga agagattaaa ccattatatg aacatcttca tgcctatgtg
     1441 agggcaaagt tgatgaatgc ctatccttcc tatatcagtc caattggatg cctccctgct
     1501 catttgcttg gtgatatgtg gggtagattt tggacaaatc tgtactcttt gacagttccc
     1561 tttggacaga aaccaaacat agatgttact gatgcaatgg tggaccaggc ctgggatgca
     1621 cagagaatat tcaaggaggc cgagaagttc tttgtatctg ttggtcttcc taatatgact
     1681 caaggattct gggaaaattc catgctaacg gacccaggaa atgttcagaa agcagtctgc
     1741 catcccacag cttgggacct ggggaagggc gacttcagga tccttatgtg cacaaaggtg
     1801 acaatggacg acttcctgac agctcatcat gagatggggc atatccagta tgatatggca
     1861 tatgctgcac aaccttttct gctaagaaat ggagctaatg aaggattcca tgaagctgtt
     1921 ggggaaatca tgtcactttc tgcagccaca cctaagcatt taaaatccat tggtcttctg
     1981 tcacccgatt ttcaagaaga caatgaaaca gaaataaact tcctgctcaa acaagcactc
     2041 acgattgttg ggactctgcc atttacttac atgttagaga agtggaggtg gatggtcttt
     2101 aaaggggaaa ttcccaaaga ccagtggatg aaaaagtggt gggagatgaa gcgagagata
     2161 gttggggtgg tggaacctgt gccccatgat gaaacatact gtgaccccgc atctctgttc
     2221 catgtttcta atgattactc attcattcga tattacacaa ggacccttta ccaattccag
     2281 tttcaagaag cactttgtca agcagctaaa catgaaggcc ctctgcacaa atgtgacatc
     2341 tcaaactcta cagaagctgg acagaaactg ttcaatatgc tgaggcttgg aaaatcagaa
     2401 ccctggaccc tagcattgga aaatgttgta ggagcaaaga acatgaatgt aaggccactg
     2461 ctcaactact ttgagccctt atttacctgg ctgaaagacc agaacaagaa ttcttttgtg
     2521 ggatggagta ccgactggag tccatatgca gaccaaagca tcaaagtgag gataagccta
     2581 aaatcagctc ttggagataa agcatatgaa tggaacgaca atgaaatgta cctgttccga
     2641 tcatctgttg catatgctat gaggcagtac tttttaaaag taaaaaatca gatgattctt
     2701 tttggggagg aggatgtgcg agtggctaat ttgaaaccaa gaatctcctt taatttcttt
     2761 gtcactgcac ctaaaaatgt gtctgatatc attcctagaa ctgaagttga aaaggccatc
     2821 aggatgtccc ggagccgtat caatgatgct ttccgtctga atgacaacag cctagagttt
     2881 ctggggatac agccaacact tggacctcct aaccagcccc ctgtttccgg atctggccac
     2941 caccaccacc accaccacca cctcgagccc aaatcttgtg acaaaactca cacatgccca
     3001 ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc cccaaaaccc
     3061 aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc
     3121 cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc
     3181 aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc
     3241 gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc
     3301 ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag
     3361 gtgtacaccc tgcccccatc ccgggatgag ctgaccaaga accaggtcag cctgacctgc
     3421 ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg
     3481 gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt cttcctctac
     3541 agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg
     3601 atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa
     3661 tgaggatccg cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc
     3721 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag
     3781 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt
     3841 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg
     3901 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc
     3961 tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat
     4021 tccgcccctc tccccccccc ccctctccct cccccccccc taacgttact ggccgaagcc
     4081 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     4141 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     4201 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     4261 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     4321 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     4381 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     4441 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     4501 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     4561 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc
     4621 acaacccggg ataattcctg cagccaatat gggatcggcc attgaacaag atggattgca
     4681 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4741 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4801 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4861 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4921 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4981 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5041 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5101 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5161 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca
     5221 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5281 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5341 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5401 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaggggatca
     5461 attctctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     5521 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     5581 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5701 ttaacgcgtc gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5761 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5821 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5881 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5941 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6001 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6061 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6121 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6841 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6901 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6961 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7021 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7081 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7141 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7201 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7261 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7321 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7381 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7441 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7501 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7561 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7621 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7681 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7741 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7801 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7861 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7921 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7981 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8041 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     8101 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     8161 cgctgcttcg cgatgtacgg
//
