LOCUS       sequence-197037-        5647 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Genetically encoded voltage indicator ASAP2s expressed under strong 
            mammalian promoter (CAG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5647)
  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 5647)
  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..5647
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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             2574..3173
                     /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(2574..2593)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3070..3090
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3333..3454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3370..3389)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3424..3443
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3503..3519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3503..3519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3516..3538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3527..3543
                     /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(3551..3557,3558..3575,3576..3581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3596..3617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3734..3751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3905..4493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3985..4004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4664..5455,5456..5524))
                     /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     5287..5306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5525..5629)
                     /gene="bla"
                     /label=AmpR promoter
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 cgcatgaccg
     2581 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     2641 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     2701 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     2761 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     2821 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     2881 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     2941 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     3001 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     3061 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     3121 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgattcgaaa
     3181 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3241 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3301 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3361 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3421 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3481 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3541 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3601 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3661 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3721 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3781 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3841 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4321 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5641 gaaaagt
//
