LOCUS       sequence-353455-        7346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric version of the genetically encoded voltage indicator 
            ASAP2s-I67T-Q397R..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7346)
  AUTHORS   Ruhl P, Langner JM, Reidel J, Schonherr R, Hoshi T, Heinemann SH
  TITLE     Monitoring of compound resting membrane potentials of cell cultures 
            with ratiometric genetically encoded voltage indicators.
  JOURNAL   Commun Biol. 2021 Oct 7;4(1):1164. doi: 10.1038/s42003-021-02675-0.
  PUBMED    34620975
REFERENCE   2  (bases 1 to 7346)
  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: "Commun 
            Biol."; date: "2021-10-7"; volume: "4(1):1164. doi"; pages: " 
            10.1038/s42003-021-02675-0"
COMMENT     Sequence Label: rASAP
FEATURES             Location/Qualifiers
     source          1..7346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2937..2954)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2943..3167
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3213..3641
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3300..3319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3510..3531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3650..3670)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3655..3984
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3835..3970
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3897..3916
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4051..4845
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4105..4124)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4715..4734
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5019..5140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5056..5075)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5110..5129
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5189..5205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5189..5205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5202..5224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5213..5229
                     /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(5237..5243,5244..5261,5262..5267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5282..5303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5420..5437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5591..6179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5671..5690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6350..7141,7142..7210))
                     /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     6973..6992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7211..7315)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atgcacatgg tcagcgagct gatcaaggaa aacatgcaca tgaaactgta catggagggg
      961 actgtgaaca atcaccattt caaatgcacc tccgagggcg aagggaagcc ctacgagggc
     1021 acacagacta tgaggatcaa ggcagtggag ggaggaccac tgccattcgc ctttgacatt
     1081 ctggctacct cattcatgta cggcagcaaa accttcatca atcacactca ggggattccc
     1141 gacttcttta agcagtcttt ccctgaaggc tttacttggg agcgagtgac cacatacgag
     1201 gatggaggcg tcctgaccgc cacacaggac acaagtctgc aggatggctg tctgatctat
     1261 aacgtgaaga ttcgcggggt caactttccc agtaatggac ctgtgatgca gaagaaaacc
     1321 ctgggatggg aggcttcaac tgaaaccctg tacccagcag acggaggact ggagggacga
     1381 gcagatatgg ctctgaaact ggtgggcggg ggacacctga tctgcaacct gaagactacc
     1441 tatcggtcca agaaacctgc taagaatctg aaaatgccag gcgtgtacta tgtggaccgg
     1501 agactggaga gaattaagga agcagataaa gagacctacg tggagcagca cgaagtggct
     1561 gtcgcacgat attgtgacct gccttctaaa ctgggccatc gggaattcat ggagacgact
     1621 gtgaggtatg aacaggggtc agagctcact aaaacttcga gctctccaac agcagatgag
     1681 cccacgataa agattgatga tggtcgtgat gagggtaatg aacaagacag ctgttccaat
     1741 accattagga gaaaaatttc cccgtttgtg atgtcatttg gattcagagt atttggagtt
     1801 gtgcttacta ttgtagacat catagtggtg attgtggatc tggccatcag tgagaagaaa
     1861 agaggcatta gagagattct tgaaggtgtt tccctggcta tagcactctt cttccttgtt
     1921 gatgttctca tgagagtgtt tgttgaaggc ttcaagaact atttccggtc caaactgaat
     1981 actttggatg cagtcatagt agtgggcact ctgctaatta atatgaccta ctccttctct
     2041 gaccttgctg cctttaacag ccataacgtg tatattaccg cggataaaca gaaaaacggc
     2101 attaaagcga actttaccgt gcgccataac gtggaagatg gcagcgtgca gctggcggat
     2161 cattatcagc agaacacccc gattggcgat ggcccggtgc tgctgccgga taaccattat
     2221 ctgagcaccc agaccgtgct gagcaaagat ccgaacgaaa aacgcgatca catggtgctg
     2281 ctggaatttg tgaccgcagc gggcattaca cacggcatgg atgaactgta tggcggcacc
     2341 ggcggcagcg cgagccaggg cgaagaactg tttaccggcg tggtgccgat tctggtggaa
     2401 ctggatggcg atgtgaacgg ccataaattt agcgtgcgcg gcgaaggcga aggcgatgcg
     2461 accattggca aactgaccct gaaatttatt tgcaccaccg gcaaactacc ggtgccgtgg
     2521 ccgaccctgg tgaccacctt aacctatggc gtgcagtgct ttagccgcta tccggatcat
     2581 atgaaacgcc atgatttttt taaaagcgcg atgccggaag gctatgtgca ggaacgcacc
     2641 attagcttta aagatgatgg caaatataaa acccgcgcgg tggtgaaatt tgaaggcgat
     2701 accctggtga accgcattga actgaaaggc accgatttta aagaagatgg caacattctg
     2761 gggcataaac tggaatataa cacagatcag atgccgcgga tggttactct tcttcgagtt
     2821 ctgagaattg ttatcttaat aagaatattt cagctggctt cacagaagaa acaacttgaa
     2881 gtggtaacct aagcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     2941 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3001 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3061 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3121 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3181 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3241 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3301 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3361 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3421 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3481 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3541 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3601 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3661 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3721 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3781 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3841 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3901 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3961 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4021 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4081 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4141 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4201 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4261 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4321 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4381 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4441 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4501 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4561 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4621 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4681 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4741 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4801 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4861 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4921 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4981 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5041 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5101 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5161 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5221 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5281 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5341 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5401 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5461 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5521 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5581 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5641 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5701 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5761 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5821 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5881 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5941 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6001 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6061 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6121 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6181 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6241 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6301 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6361 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6421 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6481 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6541 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6601 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6661 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6721 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6781 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6841 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6901 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6961 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7021 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7081 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7141 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7201 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7261 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7321 ttccccgaaa agtgccacct gacgtc
//
