LOCUS       sequence-191004-        7389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded voltage indicator ASAP2s expressed under strong 
            mammalian promoter (CAG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7389)
  AUTHORS   Chamberland S, Yang HH, Pan MM, Evans SW, Guan S, Chavarha M, Yang 
            Y, Salesse C, Wu H, Wu JC, Clandinin TR, Toth K, Lin MZ, St-Pierre F
  TITLE     Fast two-photon imaging of subcellular voltage dynamics in neuronal 
            tissue with genetically encoded indicators.
  JOURNAL   eLife. 2017 Jul 27;6. pii: e25690. doi: 10.7554/eLife.25690.
  PUBMED    28749338
REFERENCE   2  (bases 1 to 7389)
  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: "eLife."; 
            date: "2017-07-27"; volume: "6. pii: e25690. doi"; pages: " 
            10.7554/eLife.25690"
COMMENT     Sequence Label: pcDNA3.1/Puro-CAG-ASAP2s
FEATURES             Location/Qualifiers
     source          1..7389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      112..121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1516..1533)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1522..1746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1792..2220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1898)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2089..2110
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2229..2249)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2234..2563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2414..2549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2476..2495
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2572..3171
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2572..2591)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3068..3088
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3331..3452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3368..3387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3422..3441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3501..3517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3501..3517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3514..3536)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3525..3541
                     /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(3549..3555,3556..3573,3574..3579))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3594..3615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3732..3749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3903..4488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3983..4002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4659..5450,5451..5519))
                     /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     5282..5301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5520..5624)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5670..6049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6051..6324
                     /label=chicken beta-actin promoter
     intron          6325..7341
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7265..7287
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7349..7368
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gaattcctcg aggaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
       61 gtggccaatg ccctggctca caaataccac tgagatctgc atgagctagc cgccaccatg
      121 gagacgactg tgaggtatga acaggggtca gagctcacta aaacttcgag ctctccaaca
      181 gcagatgagc ccacgataaa gattgatgat ggtcgtgatg agggtaatga acaagacagc
      241 tgttccaata ccattaggag aaaaatttcc ccgtttgtga tgtcatttgg attcagagta
      301 tttggagttg tgcttatcat tgtagacatc atagtggtga ttgtggatct ggccatcagt
      361 gagaagaaaa gaggcattag agagattctt gaaggtgttt ccctggctat agcactcttc
      421 ttccttgttg atgttctcat gagagtgttt gttgaaggct tcaagaacta tttccggtcc
      481 aaactgaata ctttggatgc agtcatagta gtgggcactc tgctaattaa tatgacctac
      541 tccttctctg accttgctgc ctttaacagc cataacgtgt atattaccgc ggataaacag
      601 aaaaacggca ttaaagcgaa ctttaccgtg cgccataacg tggaagatgg cagcgtgcag
      661 ctggcggatc attatcagca gaacaccccg attggcgatg gcccggtgct gctgccggat
      721 aaccattatc tgagcaccca gaccgtgctg agcaaagatc cgaacgaaaa acgcgatcac
      781 atggtgctgc tggaatttgt gaccgcagcg ggcattacac acggcatgga tgaactgtat
      841 ggcggcaccg gcggcagcgc gagccagggc gaagaactgt ttaccggcgt ggtgccgatt
      901 ctggtggaac tggatggcga tgtgaacggc cataaattta gcgtgcgcgg cgaaggcgaa
      961 ggcgatgcga ccattggcaa actgaccctg aaatttattt gcaccaccgg caaactaccg
     1021 gtgccgtggc cgaccctggt gaccacctta acctatggcg tgcagtgctt tagccgctat
     1081 ccggatcata tgaaacgcca tgattttttt aaaagcgcga tgccggaagg ctatgtgcag
     1141 gaacgcacca ttagctttaa agatgatggc aaatataaaa cccgcgcggt ggtgaaattt
     1201 gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggca ccgattttaa agaagatggc
     1261 aacattctgg ggcataaact ggaatataac acagatcaga tgccgcagat ggttactctt
     1321 cttcgagttc tgagaattgt tatcttaata agaatatttc agctggcttc acagaagaaa
     1381 caacttgaag tggtaaccta atgaaagctt ggtaccgagc tcggatccac tagtccagtg
     1441 tggtggaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt
     1501 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1561 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1621 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1681 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     1741 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     1801 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1861 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1921 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1981 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2041 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2101 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2161 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2221 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2281 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2341 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2401 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2461 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2521 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc catgaccgag
     2581 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     2641 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     2701 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     2761 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     2821 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     2881 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     2941 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     3001 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     3061 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     3121 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg attcgaaatg
     3181 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3241 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3301 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     3361 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3421 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     3481 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3541 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3601 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3661 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3721 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3781 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3841 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3901 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3961 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4021 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4081 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4141 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4201 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4261 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4321 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4381 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4441 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4501 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4561 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4621 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4681 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4741 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4801 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4861 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4921 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4981 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5041 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5101 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5161 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5221 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5281 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5341 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5401 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5461 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5521 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5581 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5641 gtgccacctg acgtcgacgg atcggggtcg acattgatta ttgactagtt attaatagta
     5701 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5761 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5821 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5881 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5941 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6001 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     6061 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
     6121 atttattttt taattatttt gtgcagcgat gggggcgggg ggggggggcg cgcgccaggc
     6181 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
     6241 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
     6301 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
     6361 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
     6421 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
     6481 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
     6541 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
     6601 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt
     6661 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
     6721 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
     6781 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
     6841 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
     6901 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
     6961 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
     7021 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
     7081 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
     7141 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     7201 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     7261 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     7321 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     7381 tttggcaaa
//
