LOCUS       sequence-356306-        7402 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 7402)
  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 7402)
  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-VARNAM A122D WPRE-bGH-polyA
FEATURES             Location/Qualifiers
     source          1..7402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     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"
     intron          3298..3430
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3413..3432
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3439..4027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3492..3512)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3910..3921)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4054..4071)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4060..4171
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4267..4287)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4267..4285)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4306..4322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4424..4443)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4489..4544
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4543..4562)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4905..4921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4929..4945
                     /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(4953..4959,4960..4977,4978..4983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4998..5019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5078..5273
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5124..5259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5279..5413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5316..5335)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5370..5389
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5481..5498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5652..6240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5732..5751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6411..7202,7203..7271))
                     /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     7034..7053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7272..7376)
                     /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 gggggsgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc 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 atattagctg 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 ttcaagtgga ggtccggact
     2461 gattaaggaa aatatgcgga tgaaggtcgt gatggaaggg tctgtcaatg ggcaccagtt
     2521 caagtgcacc ggagagggag agggcaggcc atacgagggc gtgcagacaa tgaggatcaa
     2581 agtgatcgag ggaggaccac tgcctttcgc ctttgacatc ctggccacca gcttcatgta
     2641 cggcagcagg accttcatca agtatccagc cgacatcccc gatttcttta agcagagctt
     2701 ccccgagggc tttacctggg agagggtgac aagatacgag gatggcggcg tggtgaccgt
     2761 gacacaggac acctctctgg aggatggcga gctggtgtat aacgtgaagg tgaggggcgt
     2821 gaacttccct agcaatggcc cagtgatgca gaagaagacc aagggctggg agcccaatac
     2881 agagatgatg taccctgcag acggaggcct gaggggatat accgacatcg ccctgaaggt
     2941 ggatggagga ggacacctgc actgtaactt tgtgaccaca taccgctcca agaagacagt
     3001 gggcaatatc aagatgccag gagtgcacgc cgtggaccac aggctggagc gcatcgagga
     3061 gtctgataac gagacatatg tggtgcagag agaggtggcc gtggccaagt actctaatct
     3121 gggcggcggg atggacgagc tgtataagaa gagcaggatc accagcgagg gcgagtacat
     3181 ccccctggac cagatcgaca tcaacgtgtt ctgctacgag aacgaggtgt aagaattcga
     3241 tatcggtacc gatgaatagc tcgaggtacc gatgaatagc taaggtcgag gccgcaggta
     3301 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     3361 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
     3421 ctctccacag gtgtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3481 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3541 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3601 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3661 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3721 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3781 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     3841 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3901 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3961 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4021 tccccgcctg gaattcgagc tcggtacgat cagcctcgac tgtgccttct agttgccagc
     4081 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4141 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4201 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcccagctt
     4261 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4321 tgtgtgaaat tgttatccgc taattcactc ctcaggtgca ggctgcctat cagaaggtgg
     4381 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     4441 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     4501 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     4561 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     4621 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     4681 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     4741 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     4801 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     4861 tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     4921 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4981 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5041 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     5101 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5161 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5221 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     5281 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5341 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5401 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5461 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5521 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5581 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5641 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5701 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5761 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5821 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5881 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5941 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6001 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6061 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6121 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6181 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6241 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6301 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6541 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6601 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6661 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6721 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6781 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6841 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6901 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6961 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7021 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7081 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7141 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7201 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7261 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7321 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7381 tttccccgaa aagtgccacc tg
//
