LOCUS       sequence-357918-        8468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SNAP-fused ACE2 protein that can be fluorescently labelled
            using SNAP substrates..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8468)
  AUTHORS   Cecon E, Burridge M, Cao L, Carter L, Ravichandran R, Dam J, Jockers
            R
  TITLE     SARS-COV-2 spike binding to ACE2 in living cells monitored by 
            TR-FRET.
  JOURNAL   Cell Chem Biol. 2021 Jul 2. pii: S2451-9456(21)00307-X. doi: 
            10.1016/j.chembiol.2021.06.008.
  PUBMED    34246414
REFERENCE   2  (bases 1 to 8468)
  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 Chem 
            Biol. 2021 Jul 2. pii: S2451-9456(21)00307-X. doi: 
            10.1016/j.chembiol.2021.06.008."
COMMENT     Sequence Label: SNAP-ACE2
FEATURES             Location/Qualifiers
     source          1..8468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..31)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        203..582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        583..786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     737..757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     831..850
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        831..849
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             972..995
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1014..1559
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for mammalian expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     primer_bind     complement(4027..4044)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4033..4257
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4303..4731
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4390..4409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4600..4621
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4740..4760)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4745..5074
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4925..5060
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4987..5006
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5141..5935
                     /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(5195..5214)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5805..5824
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6109..6230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6146..6165)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6200..6219
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6279..6295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6279..6295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6292..6314)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6303..6319
                     /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(6327..6333,6334..6351,6352..6357))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6372..6393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6510..6527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6681..7269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6761..6780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7440..8231,8232..8300))
                     /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     8063..8082
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8301..8405)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
       61 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      121 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      181 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      241 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      301 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      361 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      421 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      481 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      541 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      601 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      661 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      721 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      781 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      841 cactataggg agacccaagc tggctagcgt ttaaacttaa gctttgagac atggccttac
      901 cagtgaccgc cttgctcctg ccgctggcct tgctgctcca cgccgccagg ccggccgccg
      961 ctagcggcat cgactacaag gacgacgatg acaaggccgg catcgatgcc atcatggaca
     1021 aagactgcga aatgaagcgc accaccctgg atagccctct gggcaagctg gaactgtctg
     1081 ggtgcgaaca gggcctgcac gagatcaagc tgctgggcaa aggaacatct gccgccgacg
     1141 ccgtggaagt gcctgcccca gccgccgtgc tgggcggacc agagccactg atgcaggcca
     1201 ccgcctggct caacgcctac tttcaccagc ctgaggccat cgaggagttc cctgtgccag
     1261 ccctgcacca cccagtgttc cagcaggaga gctttacccg ccaggtgctg tggaaactgc
     1321 tgaaagtggt gaagttcgga gaggtcatca gctaccagca gctggccgcc ctggccggca
     1381 atcccgccgc caccgccgcc gtgaaaaccg ccctgagcgg aaatcccgtg cccattctga
     1441 tcccctgcca ccgggtggtg tctagctctg gcgccgtggg gggctacgag ggcgggctcg
     1501 ccgtgaaaga gtggctgctg gcccacgagg gccacagact gggcaagcct gggctgggtg
     1561 atatccagca cagtggcggc cgcatctcag aggaggacct gggcagccct ggcggtgcct
     1621 ccaccattga ggaacaggcc aagacatttt tggacaagtt taaccacgaa gccgaagacc
     1681 tgttctatca aagttcactt gcttcttgga attataacac caatattact gaagagaatg
     1741 tccaaaacat gaataatgct ggggacaaat ggtctgcctt tttaaaggaa cagtccacac
     1801 ttgcccaaat gtatccacta caagaaattc agaatctcac agtcaagctt cagctgcagg
     1861 ctcttcagca aaatgggtct tcagtgctct cagaagacaa gagcaaacgg ttgaacacaa
     1921 ttctaaatac aatgagcacc atctacagta ctggaaaagt ttgtaaccca gataatccac
     1981 aagaatgctt attacttgaa ccaggtttga atgaaataat ggcaaacagt ttagactaca
     2041 atgagaggct ctgggcttgg gaaagctgga gatctgaggt cggcaagcag ctgaggccat
     2101 tatatgaaga gtatgtggtc ttgaaaaatg agatggcaag agcaaatcat tatgaggact
     2161 atggggatta ttggagagga gactatgaag taaatggggt agatggctat gactacagcc
     2221 gcggccagtt gattgaagat gtggaacata cctttgaaga gattaaacca ttatatgaac
     2281 atcttcatgc ctatgtgagg gcaaagttga tgaatgccta tccttcctat atcagtccaa
     2341 ttggatgcct ccctgctcat ttgcttggtg atatgtgggg tagattttgg acaaatctgt
     2401 actctttgac agttcccttt ggacagaaac caaacataga tgttactgat gcaatggtgg
     2461 accaggcctg ggatgcacag agaatattca aggaggccga gaagttcttt gtatctgttg
     2521 gtcttcctaa tatgactcaa ggattctggg aaaattccat gctaacggac ccaggaaatg
     2581 ttcagaaagc agtctgccat cccacagctt gggacctggg gaagggcgac ttcaggatcc
     2641 ttatgtgcac aaaggtgaca atggacgact tcctgacagc tcatcatgag atggggcata
     2701 tccagtatga tatggcatat gctgcacaac cttttctgct aagaaatgga gctaatgaag
     2761 gattccatga agctgttggg gaaatcatgt cactttctgc agccacacct aagcatttaa
     2821 aatccattgg tcttctgtca cccgattttc aagaagacaa tgaaacagaa ataaacttcc
     2881 tgctcaaaca agcactcacg attgttggga ctctgccatt tacttacatg ttagagaagt
     2941 ggaggtggat ggtctttaaa ggggaaattc ccaaagacca gtggatgaaa aagtggtggg
     3001 agatgaagcg agagatagtt ggggtggtgg aacctgtgcc ccatgatgaa acatactgtg
     3061 accccgcatc tctgttccat gtttctaatg attactcatt cattcgatat tacacaagga
     3121 ccctttacca attccagttt caagaagcac tttgtcaagc agctaaacat gaaggccctc
     3181 tgcacaaatg tgacatctca aactctacag aagctggaca gaaactgttc aatatgctga
     3241 ggcttggaaa atcagaaccc tggaccctag cattggaaaa tgttgtagga gcaaagaaca
     3301 tgaatgtaag gccactgctc aactactttg agcccttatt tacctggctg aaagaccaga
     3361 acaagaattc ttttgtggga tggagtaccg actggagtcc atatgcagac caaagcatca
     3421 aagtgaggat aagcctaaaa tcagctcttg gagataaagc atatgaatgg aacgacaatg
     3481 aaatgtacct gttccgatca tctgttgcat atgctatgag gcagtacttt ttaaaagtaa
     3541 aaaatcagat gattcttttt ggggaggagg atgtgcgagt ggctaatttg aaaccaagaa
     3601 tctcctttaa tttctttgtc actgcaccta aaaatgtgtc tgatatcatt cctagaactg
     3661 aagttgaaaa ggccatcagg atgtcccgga gccgtatcaa tgatgctttc cgtctgaatg
     3721 acaacagcct agagtttctg gggatacagc caacacttgg acctcctaac cagccccctg
     3781 tttccatatg gctgattgtt tttggagttg tgatgggagt gatagtggtt ggcattgtca
     3841 tcctgatctt cactgggatc agagatcgga agaagaaaaa taaagcaaga agtggagaaa
     3901 atccttatgc ctccatcgat attagcaaag gagaaaataa tccaggattc caaaacactg
     3961 atgatgttca gacctccttt tagtaactcg aggcctctag agggcccgtt taaacccgct
     4021 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     4081 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4141 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4201 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     4261 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     4321 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4381 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4441 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4501 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4561 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4621 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4681 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4741 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4801 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4861 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4921 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4981 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     5041 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     5101 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     5161 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5221 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5281 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5341 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5401 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5461 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5521 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5581 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5641 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5701 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5761 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5821 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5881 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5941 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     6001 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     6061 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     6121 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6181 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6241 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6301 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6361 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6421 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6481 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6541 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6601 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6661 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6721 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6781 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6841 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6901 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6961 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7021 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7081 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     7141 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7201 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7261 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7321 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7381 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7441 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7501 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7561 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7621 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7681 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7741 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7801 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7861 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7921 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7981 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8041 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8101 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8161 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8221 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8281 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8341 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8401 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     8461 ccctatgg
//
