LOCUS       sequence-340941-        7637 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of voltage sensor; soma localization targeting 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7637)
  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 7637)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-ST A122D-WPRE-bGH-polyA
FEATURES             Location/Qualifiers
     source          1..7637
                     /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          3559..3691
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3674..3693
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3700..4288
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3753..3773)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4171..4182)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4315..4332)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4321..4432
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4528..4548)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4528..4546)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4659..4678)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4724..4779
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4778..4797)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5140..5156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5140..5156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5153..5175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5164..5180
                     /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(5188..5194,5195..5212,5213..5218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5233..5254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5313..5508
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5359..5494
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5514..5648
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5551..5570)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5605..5624
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5716..5733)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5887..6475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5967..5986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6646..7437,7438..7506))
                     /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     7269..7288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7507..7611)
                     /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 gccrggsggg gggggggggg ggggggggsg
      541 gggsggggsg rggsggrgag 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 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 ggtgaagagc aggatcacca gcgagggcga
     3241 gtacatcccc ctggaccaga tcgacatcaa cgtgttctgc tacgagaacg aggtgcaaag
     3301 tcagcctatc ctgaacacaa aggaaatggc tccacagtct aagcctcccg aagagcttga
     3361 gatgtccagt atgccaagtc ccgtggctcc cctccctgcc aggactgaag gagtgattga
     3421 catgaggagt atgtcatcta ttgatagctt catctcttgc gcaacagatt tccccgaggc
     3481 tactcgattc tgagaattcg atatcggtac cgatgaatag ctcgaggtac cgatgaatag
     3541 ctaaggtcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
     3601 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
     3661 actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa
     3721 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3781 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3841 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3901 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3961 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4021 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4081 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4141 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4201 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4261 cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga
     4321 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4381 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4441 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4501 gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     4561 atcatggtca tagcykwwty cwstsykmar ktgywrkctg cctatcagaa ggtggtggct
     4621 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4681 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4741 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4801 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     4861 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     4921 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     4981 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     5041 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     5101 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5161 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5221 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5281 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     5341 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5401 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5461 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     5521 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5581 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5641 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5701 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5761 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5821 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5881 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5941 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6001 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6061 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6121 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6181 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6241 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6301 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6361 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6421 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6481 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6541 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6601 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6661 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6721 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6781 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6841 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6901 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6961 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7021 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7081 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7141 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7201 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7261 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7321 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7381 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7441 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7501 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7561 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7621 ccgaaaagtg ccacctg
//
