LOCUS       sequence-254336-        7663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express soma-localized Ace2_D92N_E199V-mNeon in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7663)
  AUTHORS   Abdelfattah AS, Valenti R, Zheng J, Wong A, Podgorski K, Koyama M, 
            Kim DS, Schreiter ER
  TITLE     A general approach to engineer positive-going eFRET voltage 
            indicators.
  JOURNAL   Nat Commun. 2020 Jul 10;11(1):3444. doi: 10.1038/s41467-020-17322-1.
  PUBMED    32651384
REFERENCE   2  (bases 1 to 7663)
  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: "Nat 
            Commun."; date: "2020-07-10"; volume: "11(1):3444. doi"; pages: " 
            10.1038/s41467-020-17322-1"
COMMENT     Sequence Label: pCAG-Ace2_D92N_E199V-mNeon-ST
FEATURES             Location/Qualifiers
     source          1..7663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      8..17
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      929..938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          1807..1939
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1922..1941
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1948..2536
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2001..2021)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2419..2430)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2563..2580)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2569..2680
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2776..2796)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2776..2794)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2815..2831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2952)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2998..3053
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3052..3071)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3414..3430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3438..3454
                     /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(3462..3468,3469..3486,3487..3492))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3507..3528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3587..3782
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3633..3768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3788..3922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3825..3844)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3879..3898
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3990..4007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4161..4749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4241..4260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4920..5711,5712..5780))
                     /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     5543..5562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5781..5885)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5916..6295
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6573..7589
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7513..7535
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7597..7616
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 cggtgccgcc accatggctg acgtggaaac cgagaccggc atgattgcac agtggattgt
       61 ctttgctatt atggctgctg ctgctattgc ttttggagtg gctgtgcact ttcggccttc
      121 agagctgaag agcgcatact atatcaacat tgccatctgc actatcgccg ctaccgctta
      181 ctatgcaatg gccgtgaact accaggacct gacaatgaat ggtgaaaggc aggtggtcta
      241 cgcaagatat attgactggg tgctgaccac accactgctc ctgctcaacc tcatcgtcat
      301 gaccaagatg ggcggagtga tgatttcttg ggtcatcggc gcagacattt tcatgatcgt
      361 gtttggtatt ctgggcgcct tcgaggatga acacaagttc aaatgggtgt actttatcgc
      421 tggatgtgtg atgcaggcag tcctgacata cgggatgtat aacgccactt ggaaagacga
      481 tctgaagaaa agccccgagt accatagctc ctatgtcagt ctgctcgtct tcctgtcaat
      541 cctctgggtg ttttatcctg tcgtgtgggc tttcgggtct ggtagtggcg tgctgtccgt
      601 cgacaatgtg gccattctca tgggaatcct ggatgtgctc gctaagccac tgtttggaat
      661 ggggtgcctc attgcccatg agactatctt caagaagatg ctgaggtctc tcccagcgac
      721 acatgagtta cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg
      781 caccggcaat ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct
      841 ccagttctcc ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc
      901 ctaccctgac gggatgtcgc ctttccaggc cgccatggta gatggctccg gataccaagt
      961 ccatcgcaca atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta
     1021 cgagggaagc cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg
     1081 tcctgtgatg accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc
     1141 caacgacaaa accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg
     1201 ctacaggagc actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct
     1261 gaagaaccag ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct
     1321 caacttcaag gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa
     1381 gaagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg acatcaacgt
     1441 gttctgctac gagaacgagg tgaagagcag gatcaccagc gagggcgagt acatccccct
     1501 ggaccagatc gacatcaacg tgttctgcta cgagaacgag gtgcaaagtc agcctatcct
     1561 gaacacaaag gaaatggctc cacagtctaa gcctcccgaa gagcttgaga tgtccagtat
     1621 gccaagtccc gtggctcccc tccctgccag gactgaagga gtgattgaca tgaggagtat
     1681 gtcatctatt gatagcttca tctcttgcgc aacagatttc cccgaggcta ctcgattctg
     1741 agaattcgat atcggtaccg atgaatagct cgaggtaccg atgaatagct aaggtcgagg
     1801 ccgcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     1861 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     1921 tttgcctttc tctccacagg tgtcgacaat caacctctgg attacaaaat ttgtgaaaga
     1981 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2041 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2101 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2161 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2221 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2281 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2341 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2401 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2461 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     2521 tgggccgcct ccccgcctgg aattcgagct cggtacgatc agcctcgact gtgccttcta
     2581 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2641 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2701 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2761 gcccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2821 gctgtttcct gtgtgaaatt gttatccgct aattcactcc tcaggtgcag gctgcctatc
     2881 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     2941 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     3001 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     3061 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     3121 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     3181 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3241 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3301 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3361 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     3421 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3481 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3541 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     3601 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3661 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3721 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3781 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3841 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3901 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     3961 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4021 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4081 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4201 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4261 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4321 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4381 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4501 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4561 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4621 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4681 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4741 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4801 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4861 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4921 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4981 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5041 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5101 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5161 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5221 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5281 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5341 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5401 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5461 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5521 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5581 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5641 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5701 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5761 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5821 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5881 ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga ctagttatta
     5941 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     6001 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6061 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     6121 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     6181 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6241 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     6301 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     6361 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggsgcg
     6421 cgccrggsgg ggsggggsgg gggggggggs ggggsggggs gaggcggaga ggtgcggcgg
     6481 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
     6541 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
     6601 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
     6661 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat
     6721 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt
     6781 tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt
     6841 gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc
     6901 tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct
     6961 gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg
     7021 gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc
     7081 cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg
     7141 ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg
     7201 gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc
     7261 attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg
     7321 cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt
     7381 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
     7441 cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac
     7501 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
     7561 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     7621 ctcatcattt tggcaaagaa ttcggtaccg cgggcccggg aac
//
