LOCUS       sequence-383234-        7101 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Flp recombinase-dependent red fluorescent, negative 
            response-polarity voltage indicator; soma-targeted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7101)
  AUTHORS   Kannan M, Vasan G, Haziza S, Huang C, Chrapkiewicz R, Luo J, Cardin 
            JA, Schnitzer MJ, Pieribone VA
  TITLE     Dual-polarity voltage imaging of the concurrent dynamics of multiple
            neuron types.
  JOURNAL   Science. 2022 Nov 4;378(6619):eabm8797. doi: 
            10.1126/science.abm8797. Epub 2022 Nov 4.
  PUBMED    36378956
REFERENCE   2  (bases 1 to 7101)
  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: "Science."; 
            date: "2022-11-4"; volume: "378(6619):eabm8797. doi"; pages: " 
            10.1126/science.abm8797. Epub 2022 Nov 4"
COMMENT     Sequence Label: pAAV-CAG-fDIO-VARNAM2-Kv2.1PR
FEATURES             Location/Qualifiers
     source          1..7101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        210..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..792
                     /label=chicken beta-actin promoter
     primer_bind     1122..1144
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1206..1225
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1292..1325
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      3070..3079
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3086..3119)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3228..3816
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3281..3301)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3699..3710)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3848..4324
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4047..4066)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4364..4504
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4364..4493
                     /label=AAV2 ITR
     rep_origin      4579..5034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4666..4685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4876..4897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5051..5070)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5170..5192
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5230..5248)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5316..5420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5421..5489,5490..6281)
                     /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     complement(5639..5658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6452..7040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6941..6960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaacact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg
      661 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga
     1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa
     1261 cggtaccgga tcctctagag tcgactccgg agaagttcct attctctaga aagtatagga
     1321 acttcgcaga atggtagctg gattgtagct gctattagca atatgaaacc tcttagaagt
     1381 tcctattctt caaaaggtat aggaacttcg gcgcgcctta gaatcgagta gcctcgggga
     1441 aatctgttgc gcaagagatg aagctatcaa tagatgacat actcctcatg tcaatcactc
     1501 cttcagtcct ggcagggagg ggagccacgg gacttggcat actggacatc tcaagctctt
     1561 cgggaggctt agactgtgga gccatttcct ttgtgttcag gataggctga ctttgcacct
     1621 cgttctcgta gcagaacacg ttgatgtcga tctggtccag ggggatgtac tcgccctcgc
     1681 tggtgatcct gctcttctta tacagctcgt ccatcccgcc gcccagatta gagtacttgg
     1741 ccacggccac ctctctctgc accacatatg tctcgttatc agactcctcg atgcgctcca
     1801 gcctgtggtc cacggcgtgc actcctggca tcttgatatt gcccactgtc ttcttggagc
     1861 ggtatgtggt cacaaagtta cagtgcaggt gtcctcctcc atccaccttc agggcgatgt
     1921 cggtatatcc cctcaggcct ccgtctgcag ggtacatcat ctctgtattg ggctcccagc
     1981 ccttggtctt cttctgcatc actgggccat tgctagggaa gttcacgccc ctcaccttca
     2041 cgttatacac cagctcgcca tcctccagag aggtgtcctg tgtcacggtc accacgccgc
     2101 catcctcgta tcttgtcacc ctctcccagg taaagccctc ggggaagctc tgcttaaaga
     2161 aatcggggat gtcggctgga tacttgatga aggtcctgct gccgtacatg aagctggtgg
     2221 ccaggatgtc aaaggcgaaa ggcagtggtc ctccctcgat cactttgatc ctcattgtct
     2281 gcacgccctc gtatggcctg ccctctccct ctccggtgca cttgaactgg tgcccattga
     2341 cagacccttc catcacgacc ttcatccgca tattttcctt aatcaggatg gacttgaaga
     2401 tagtctcatg ggcaatgagg caccccattc caaacagtgg cttagcgagc acatccagga
     2461 ttcccatgag aatggcctca ttgtcgacgg acagcacgcc actaccagac ccgaaagccc
     2521 acacgacagg ataaaacacc cagaggattg acaggaagac gagcagactg acataggagc
     2581 tatggtactc ggggcttttc ttcagatcgt ctttccaagt ggcgttatac atcccgtatg
     2641 tcaggactgc ctgcatcaca catccagcga taaagtacac ccatttgaac ttgtgttcat
     2701 cctcgaaggc gcccagaata ccaaacacga tcatgaaaat gtctgcgccg atgacccaag
     2761 aaatcatcac tccgcccatc ttggtcatga cgatgagatc gagcaggagc agtggtgtgg
     2821 tcagcaccca gctaatatat cttgcgtaga ccacctgcct ttcaccattc attgtcaggt
     2881 cctggtagtt cacggccatt gcatagtaag cggtagcggc gatagtgcag atggcaatgt
     2941 tgatatagta tgcgctcttc agctctgaag gccgaaagtg cacagccact ccaaaagcaa
     3001 tagcagcagc agccataata gcaaagacaa tccactgtgc aatcatgccg gtctcggttt
     3061 ccacgtcagc catggtggcg ctagcgaagt tcctatactt tctagagaat aggaacttct
     3121 tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagag aagttcctat
     3181 accttttgaa gaataggaac ttcgaattcg atatcaagct tatcgataat caacctctgg
     3241 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3301 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3361 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3421 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3481 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3541 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3601 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     3661 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3721 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3781 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga
     3841 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     3901 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     3961 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     4021 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     4081 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     4141 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     4201 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     4261 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     4321 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     4381 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4441 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4501 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4561 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4621 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4681 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4741 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4801 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4861 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4921 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4981 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5041 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5101 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5161 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5221 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5281 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5341 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5401 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5461 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5521 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5581 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5641 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5701 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5761 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5821 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5881 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5941 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6001 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6061 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6121 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6181 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6241 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6301 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6361 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6421 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6481 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6541 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6601 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6661 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6721 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6781 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6841 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6901 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6961 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7021 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7081 gctggccttt tgctcacatg t
//
