LOCUS       sequence-341111-        7361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of voltage sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7361)
  AUTHORS   Abdelfattah AS, Zheng J, Singh A, Huang YC, Reep D, Tsegaye G, Tsang
            A, Arthur BJ, Rehorova M, Olson CVL, Shuai Y, Zhang L, Fu TM, Milkie
            DE, Moya MV, Weber TD, Lemire AL, Baker CA, Falco N, Zheng Q, Grimm 
            JB, Yip MC, Walpita D, Chase M, Campagnola L, Murphy GJ, Wong AM, 
            Forest CR, Mertz J, Economo MN, Turner GC, Koyama M, Lin BJ, Betzig 
            E, Novak O, Lavis LD, Svoboda K, Korff W, Chen TW, Schreiter ER, 
            Hasseman JP, Kolb I
  TITLE     Sensitivity optimization of a rhodopsin-based fluorescent voltage 
            indicator.
  JOURNAL   Neuron. 2023 Mar 28:S0896-6273(23)00205-2. doi: 
            10.1016/j.neuron.2023.03.009.
  PUBMED    37015225
REFERENCE   2  (bases 1 to 7361)
  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: "Neuron. 
            2023 Mar 28:S0896-6273(23)00205-2. doi: 
            10.1016/j.neuron.2023.03.009."
COMMENT     Sequence Label: pGP-CAG-Ace2N-4AA-mNeon A122D-WPRE-bGH-polyA
FEATURES             Location/Qualifiers
     source          1..7361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1760..1769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2681..2690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3283..3415
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3398..3417
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3424..4012
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3477..3497)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3895..3906)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4039..4056)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4045..4156
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4252..4272)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4252..4270)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4383..4402)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4448..4503
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4502..4521)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4864..4880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4864..4880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4877..4899)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4888..4904
                     /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(4912..4918,4919..4936,4937..4942))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4957..4978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5037..5232
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5083..5218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5238..5372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5275..5294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5329..5348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5440..5457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5611..6199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5691..5710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6370..7161,7162..7230))
                     /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     6993..7012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7231..7335)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccrgggggg gsgggggggg gsggggggsg
      541 gggsggggsg rggcggrgag gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccgc
     1741 gggcccggga accggtgccg ccaccatggc tgacgtggaa accgagaccg gcatgattgc
     1801 acagtggatt gtctttgcta ttatggctgc tgctgctatt gcttttggag tggctgtgca
     1861 ctttcggcct tcagagctga agagcgcata ctatatcaac attgccatct gcactatcgc
     1921 cgctaccgct tactatgcaa tggccgtgaa ctaccaggac ctgacaatga atggtgaaag
     1981 gcaggtggtc tacgcaagat atattaactg ggtgctgacc acaccactgc tcctgctcga
     2041 tctcatcgtc atgaccaaga tgggcggagt gatgatttct tgggtcatcg gcgcagacat
     2101 tttcatgatc gtgtttggta ttctgggcga cttcgaggat gaacacaagt tcaaatgggt
     2161 gtactttatc gctggatgtg tgatgcaggc agtcctgaca tacgggatgt ataacgccac
     2221 ttggaaagac gatctgaaga aaagccccga gtaccatagc tcctatgtca gtctgctcgt
     2281 cttcctgtca atcctctggg tgttttatcc tgtcgtgtgg gctttcgggt ctggtagtgg
     2341 cgtgctgtcc gtcgacaatg aggccattct catgggaatc ctggatgtgc tcgctaagcc
     2401 actgtttgga atggggtgcc tcattgccca tgagactatc ttcaagaaga tgctgaggtc
     2461 tctcccagcg acacatgagt tacacatctt tggctccatc aacggtgtgg actttgacat
     2521 ggtgggtcag ggcaccggca atccaaatga tggttatgag gagttaaacc tgaagtccac
     2581 caagggtgac ctccagttct ccccctggat tctggtccct catatcgggt atggcttcca
     2641 tcagtacctg ccctaccctg acgggatgtc gcctttccag gccgccatgg tagatggctc
     2701 cggataccaa gtccatcgca caatgcagtt tgaagatggt gcctccctta ctgttaacta
     2761 ccgctacacc tacgagggaa gccacatcaa aggagaggcc caggtgaagg ggactggttt
     2821 ccctgctgac ggtcctgtga tgaccaactc gctgaccgct gcggactggt gcaggtcgaa
     2881 gaagacttac cccaacgaca aaaccatcat cagtaccttt aagtggagtt acaccactgg
     2941 aaatggcaag cgctacagga gcactgcgcg gaccacctac acctttgcca agccaatggc
     3001 ggctaactat ctgaagaacc agccgatgta cgtgttccgt aagacggagc tcaagcactc
     3061 caagaccgag ctcaacttca aggagtggca aaaggccttt accgatgtga tgggcatgga
     3121 cgagctgtac aagaagagca ggatcaccag cgagggcgag tacatccccc tggaccagat
     3181 cgacatcaac gtgttctgct acgagaacga ggtgtaagaa ttcgatatcg gtaccgatga
     3241 atagctcgag gtaccgatga atagctaagg tcgaggccgc aggtaagtat caaggttaca
     3301 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
     3361 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc
     3421 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3481 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3541 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3601 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3661 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3721 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3781 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     3841 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3901 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3961 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     4021 cgagctcggt acgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4081 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4141 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4201 ggcaggacag caagggggag gattgggaag acaatagccc agcttttgtt ccctttagtg
     4261 agggttaatt gcgcgcttgg cgtaatcatg gtcatmssyk wwtycwstsy kmarktgcar
     4321 kctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     4381 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     4441 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     4501 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     4561 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     4621 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     4681 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     4741 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     4801 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc
     4861 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4921 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4981 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     5041 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5161 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5221 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5281 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5341 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     5401 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5461 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5521 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5581 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5641 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5701 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5761 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5821 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5881 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5941 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6001 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6061 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6121 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6181 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6241 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6301 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6361 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6421 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6481 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6541 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6601 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6661 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6721 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6781 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6841 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6901 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6961 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7021 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7081 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7141 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7201 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7261 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7321 aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
