LOCUS       sequence-259835-        7387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Ace2_D92N_E199V-mNeon in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7387)
  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 7387)
  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
FEATURES             Location/Qualifiers
     source          1..7387
                     /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          1531..1663
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1646..1665
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1672..2260
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1725..1745)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2143..2154)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2287..2304)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2293..2404
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2500..2520)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2500..2518)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2539..2555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2657..2676)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2722..2777
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2776..2795)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3138..3154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3162..3178
                     /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(3186..3192,3193..3210,3211..3216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3231..3252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3311..3506
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3357..3492
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3512..3646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3549..3568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3603..3622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3714..3731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3885..4473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3965..3984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4644..5435,5436..5504))
                     /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     5267..5286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5505..5609)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5640..6019
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6297..7313
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7237..7259
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7321..7340
                     /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 tgtaagaatt cgatatcggt accgatgaat agctcgaggt
     1501 accgatgaat agctaaggtc gaggccgcag gtaagtatca aggttacaag acaggtttaa
     1561 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     1621 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcga caatcaacct
     1681 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1741 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1801 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     1861 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     1921 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     1981 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2041 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2101 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2161 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2221 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
     2281 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2341 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2401 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2461 agggggagga ttgggaagac aatagcccag cttttgttcc ctttagtgag ggttaattgc
     2521 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctaattca
     2581 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     2641 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     2701 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     2761 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     2821 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     2881 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     2941 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     3001 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     3061 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     3121 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3181 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3241 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3301 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3361 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3421 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3481 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     3541 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3601 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     3661 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3721 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3781 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4081 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4321 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4621 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4681 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4741 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4801 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4861 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4921 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4981 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5041 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5101 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5161 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5221 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5281 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5341 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5401 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5461 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5581 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg
     5641 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     5701 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     5761 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5821 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     5881 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     5941 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6001 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     6061 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     6121 gggggcgggg gggggggggg sgcgcgccrg gsgggggggg gggggggggg ggggggggsg
     6181 gggsgrggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
     6241 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
     6301 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
     6361 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     6421 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
     6481 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt
     6541 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg
     6601 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg
     6661 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     6721 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc
     6781 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg
     6841 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
     6901 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
     6961 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     7021 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
     7081 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
     7141 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     7201 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     7261 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     7321 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcggt accgcgggcc
     7381 cgggaac
//
