LOCUS       sequence-402752-        7380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of the fluorescent voltage sensor ASAP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  AUTHORS   St-Pierre F, Marshall JD, Yang Y, Gong Y, Schnitzer MJ, Lin MZ
  TITLE     High-fidelity optical reporting of neuronal electrical activity with
            an ultrafast fluorescent voltage sensor.
  JOURNAL   Nat Neurosci. 2014 Apr 22. doi: 10.1038/nn.3709.
  PUBMED    24755780
REFERENCE   2  (bases 1 to 7380)
  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: "Nat 
            Neurosci. 2014 Apr 22. doi: 10.1038/nn.3709."
COMMENT     Sequence Label: pcDNA3.1/Puro-CAG-ASAP1
FEATURES             Location/Qualifiers
     source          1..7380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      8..17
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1412..1429)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1418..1642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1688..2116
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1775..1794)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1985..2006
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2125..2145)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2130..2459
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2310..2445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2372..2391
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2470..3069
                     /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(2470..2489)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2966..2986
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3229..3350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3266..3285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3320..3339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3399..3415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3399..3415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3412..3434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3423..3439
                     /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(3447..3453,3454..3471,3472..3477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3492..3513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3630..3647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3801..4389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3881..3900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4560..5351,5352..5420))
                     /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     5183..5202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5421..5525)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5557..5936
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6212..7228
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7152..7174
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7236..7255
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gctagccgcc accatggaga cgactgtgag gtatgaacag gggtcagagc tcactaaaac
       61 ttcgagctct ccaacagcag atgagcccac gataaagatt gatgatggtc gtgatgaggg
      121 taatgaacaa gacagctgtt ccaataccat taggagaaaa atttccccgt ttgtgatgtc
      181 atttggattc agagtatttg gagttgtgct tatcattgta gacatcatag tggtgattgt
      241 ggatctggcc atcagtgaga agaaaagagg cattagagag attcttgaag gtgtttccct
      301 ggctatagca ctcttcttcc ttgttgatgt tctcatgaga gtgtttgttg aaggcttcaa
      361 gaactatttc cggtccaaac tgaatacttt ggatgcagtc atagtagtgg gcactctgct
      421 aattaatatg acctactcct tctctgacct tgctgccttt aacagccata acgtgtatat
      481 taccgcggat aaacagaaaa acggcattaa agcgaacttt accgtgcgcc ataacgtgga
      541 agatggcagc gtgcagctgg cggatcatta tcagcagaac accccgattg gcgatggccc
      601 ggtgctgctg ccggataacc attatctgag cacccagacc gtgctgagca aagatccgaa
      661 cgaaaaacgc gatcacatgg tgctgctgga atttgtgacc gcagcgggca ttacacacgg
      721 catggatgaa ctgtatggcg gcaccggcgg cagcgcgagc cagggcgaag aactgtttac
      781 cggcgtggtg ccgattctgg tggaactgga tggcgatgtg aacggccata aatttagcgt
      841 gcgcggcgaa ggcgaaggcg atgcgaccat tggcaaactg accctgaaat ttatttgcac
      901 caccggcaaa ctaccggtgc cgtggccgac cctggtgacc accttaacct atggcgtgca
      961 gtgctttagc cgctatccgg atcatatgaa acgccatgat ttttttaaaa gcgcgatgcc
     1021 ggaaggctat gtgcaggaac gcaccattag ctttaaagat gatggcaaat ataaaacccg
     1081 cgcggtggtg aaatttgaag gcgataccct ggtgaaccgc attgaactga aaggcaccga
     1141 ttttaaagaa gatggcaaca ttctggggca taaactggaa tataacacag atcagatgcc
     1201 gcagatggtt actcttcttc gagttctgag aattgttatc ttaataagaa tatttcgcct
     1261 ggcttcacag aagaaacaac ttgaagtggt aacctaatga aagcttggta ccgagctcgg
     1321 atccactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag
     1381 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     1441 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1501 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1561 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     1621 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     1681 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1741 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1801 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1861 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1921 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1981 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2041 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2101 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2161 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2221 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2281 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2341 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     2401 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2461 agctcccgca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     2521 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     2581 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     2641 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     2701 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     2761 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     2821 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     2881 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     2941 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     3001 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     3061 ggtgcctgat tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3121 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3181 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     3241 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3301 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     3361 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     3421 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3481 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3541 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3601 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3661 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3721 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4201 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4321 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4381 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4441 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4501 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4561 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4621 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4681 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4741 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4801 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4861 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4921 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4981 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5041 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5101 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5161 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5221 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5281 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5341 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5401 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5461 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5521 ccgcgcacat ttccccgaaa agtgccacct gacgtcgaca ttgattattg actagttatt
     5581 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     5641 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     5701 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     5761 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     5821 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     5881 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     5941 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     6001 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggsgc
     6061 gccrggsggg gggggggggg ggggggggsg gggsggggsg aggsggagag gtgsggcggc
     6121 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     6181 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
     6241 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     6301 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
     6361 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
     6421 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     6481 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
     6541 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg
     6601 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     6661 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc
     6721 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg
     6781 gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg rgggctcggg
     6841 ggrggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
     6901 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc
     6961 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg
     7021 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
     7081 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
     7141 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     7201 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     7261 tcatcatttt ggcaaagaat tcctcgagga attcactcct caggtgcagg ctgcctatca
     7321 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctgcatga
//
