LOCUS       sequence-235532-        9293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ubiqutously express ASAP1 voltage biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9293)
  AUTHORS   Silic MR, Zhang G
  TITLE     Visualization of Cellular Electrical Activity in Zebrafish Early 
            Embryos and Tumors.
  JOURNAL   J Vis Exp. 2018 Apr 25;(134). doi: 10.3791/57330.
  PUBMED    29757272
REFERENCE   2  (bases 1 to 9293)
  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: "J Vis 
            Exp."; date: "2018-04-25"; volume: "(134). doi"; pages: " 
            10.3791/57330"
COMMENT     Sequence Label: pDestTol2-ubi-ASAP1
FEATURES             Location/Qualifiers
     source          1..9293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2611..2635
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3926..3950)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3952..4086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(4149..4169)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(4177..4193)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4209..4343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(4782..4969,4970..4981)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5015..5031)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5015..5031)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5068..5088)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5068..5086)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5107..5123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5120..5142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5131..5147
                     /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(5155..5161,5162..5179,5180..5185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5200..5221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5338..5355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5509..6097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5589..5608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6268..7059,7060..7128))
                     /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     6891..6910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7129..7233)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7396..7417)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(7766..7777,7778..7940)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     8313..8329
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8349..8369
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 tgaaaacatc ctcctgtcat gtatttgcta tattcaaatg ttaaacccgt gcagaataga
       61 acatatacaa aaaaaaacaa cacaacacat ttttaaacat tattaaatat caagtattgc
      121 tggcagttct gtttctgttt tacagtaccc tttgccacag ttctccgctt ttcctggtcc
      181 agattccaca agtctgattc accaatagca aagcgaataa acaaccaaag cagccaatca
      241 ctgcttgtag actgtcctgc gagaccggcc cattccagca cattctggaa acttccttta
      301 tatgataatt ataaatacat ttaaattatt gatacaaaac atgtaattcc tagaacataa
      361 ccatagcaat cattagtttt cagggtaatt atgtattttt aggatttgac tgcggaaaga
      421 tctggtcatg tgacgtctca tgaacgtcac ggccctgggt ttctataaat acagtaggac
      481 tctcgaccat cggcagattt ttcgaagaag aagatcagtt tcaggagccg tactgttccg
      541 tttcaacgca aatattaacg gtaagagcga atttcctagt ttgttttcat gccattcttt
      601 aaaaccatag cgtattactt taattatagt aaactttcgc tttctttatt acaagagacg
      661 ttttgtgttg attctccgcg gacattttcg gtcagacaat cagaaaatga ccgcggagga
      721 ccagtaactt gcattacacg taagttaaat cttcgtgtat taaaatggtt aggttgttaa
      781 cgtcaaatag gttaccgtgt ttgcgtgtga tcaggttggt tttgttagat ttttgtcagt
      841 atttttaatt tatttgtttt agtttattta ttttttttgc tgaatcatag tttgtgaaca
      901 aagaacccgg atgttacata cagtacagcc gccatgttac agagagttat aacttaatca
      961 ttttaaaaat aattttgcct tacttttagt ttgtcatgtt gagaaatgag gaaatgttaa
     1021 aatgaggaaa tatccaatta atttaatata tcaaaataat ccatgattac aatgcactga
     1081 actggagaaa attaagatgt tttctagtgt catgaaacaa atgtaagaga tgtacattgt
     1141 agatgtttta tgtcaagaat tggctagttg atgcagcata ctggcgatac tcagttgtaa
     1201 taacagtaac gttacatgtt aatagactac tgagtatgct gttctgtcta tgtatgctct
     1261 gtaagctacg agaaggactt ttttaaacag taaagggtgc aatattttta caaattgaat
     1321 taaataaagg ctgtctatta agtaatatgc ttgatatttt tcttacttga tcggaaataa
     1381 gaaaaaatat aaacgttgtt gctctaaaaa tcctagttca gtttagccaa ccacaaatac
     1441 ctttttgttc ctccaacagt ttttttttct tctctataat atttggcagt ctatagtact
     1501 ccaaatgttt ccccacagtc taactaattg gtacagccaa aatcatgaca cttattgcaa
     1561 taataatttt ggttcattgg cattgttgat agcctgtgcc actaatatgg tcgattgatc
     1621 atgcttcagg aagaaaacta tattgtttga tgtaagatta ttaaatcttc acctgcttcc
     1681 attacaaact attcccatct tattgaattc tggtatgtct taaaggatta gttcacttcc
     1741 caaatcaaaa tttacttagc cttttttcat ccatgatccc tttttttcat cattaatgaa
     1801 gaaattgttt ttgaaaaagt ttcaagattt ttttctctat attgtggagc ttgttagttt
     1861 aaaattccaa aatgcaatat gtggcttcaa atggttctaa atgatcccag tcaaggaata
     1921 acagtcttat ctaatgaaac cattagacct ttaaaaataa aaaaaataaa agtatttatt
     1981 tttaaatgac tgagtgatta agttgaattt cagcgtttcc ttactgtgta gaagtccttc
     2041 cttactggcc ccaccctttg gttctctgcc aatctgctac ctaatgtaat gttgtggaac
     2101 attattattc tttatttctt aattttttat tttttatttt aaaacaatgt aaactgcaca
     2161 gatgtgcagt ttgtttaaaa tggccaatgc tttggaaatg catgacataa ttagatttca
     2221 tgatgcacaa agccaaatct cagagcttgt gcaaaatgag ctatcatttc actaggtaag
     2281 accctaaatt ttcatatagg atcatttgga caattttgct gcaggtaaaa tgcattctat
     2341 agtccactgt cagccattgt tttggatagt atttattttt ctctacaagt atagtcaata
     2401 gttttctatt attttaaagg tttgtaacat ttaagggtga ccaaatgcaa agtaaaattt
     2461 cattttcggg tgaactatct cgtttaacat gggagaagtg caaaacatac attattggct
     2521 agaacattgt agtatttttt aaatggaaat gtgtgattgc taatcttact ttgaatttgt
     2581 ttacagggat cctttaaggg cgaattcgac ccaagtttgt acaaaaaagc aggcttcacc
     2641 atggagacga ctgtgaggta tgaacagggg tcagagctca ctaaaacttc gagctctcca
     2701 acagcagatg agcccacgat aaagattgat gatggtcgtg atgagggtaa tgaacaagac
     2761 agctgttcca ataccattag gagaaaaatt tccccgtttg tgatgtcatt tggattcaga
     2821 gtatttggag ttgtgcttat cattgtagac atcatagtgg tgattgtgga tctggccatc
     2881 agtgagaaga aaagaggcat tagagagatt cttgaaggtg tttccctggc tatagcactc
     2941 ttcttccttg ttgatgttct catgagagtg tttgttgaag gcttcaagaa ctatttccgg
     3001 tccaaactga atactttgga tgcagtcata gtagtgggca ctctgctaat taatatgacc
     3061 tactccttct ctgaccttgc tgcctttaac agccataacg tgtatattac cgcggataaa
     3121 cagaaaaacg gcattaaagc gaactttacc gtgcgccata acgtggaaga tggcagcgtg
     3181 cagctggcgg atcattatca gcagaacacc ccgattggcg atggcccggt gctgctgccg
     3241 gataaccatt atctgagcac ccagaccgtg ctgagcaaag atccgaacga aaaacgcgat
     3301 cacatggtgc tgctggaatt tgtgaccgca gcgggcatta cacacggcat ggatgaactg
     3361 tatggcggca ccggcggcag cgcgagccag ggcgaagaac tgtttaccgg cgtggtgccg
     3421 attctggtgg aactggatgg cgatgtgaac ggccataaat ttagcgtgcg cggcgaaggc
     3481 gaaggcgatg cgaccattgg caaactgacc ctgaaattta tttgcaccac cggcaaacta
     3541 ccggtgccgt ggccgaccct ggtgaccacc ttaacctatg gcgtgcagtg ctttagccgc
     3601 tatccggatc atatgaaacg ccatgatttt tttaaaagcg cgatgccgga aggctatgtg
     3661 caggaacgca ccattagctt taaagatgat ggcaaatata aaacccgcgc ggtggtgaaa
     3721 tttgaaggcg ataccctggt gaaccgcatt gaactgaaag gcaccgattt taaagaagat
     3781 ggcaacattc tggggcataa actggaatat aacacagatc agatgccgca gatggttact
     3841 cttcttcgag ttctgagaat tgttatctta ataagaatat ttcgcctggc ttcacagaag
     3901 aaacaacttg aagtggtaac ctaagaccca gctttcttgt acaaagtggg ggatccagac
     3961 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     4021 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     4081 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     4141 gttttttcca actttattat acatagttga taattcactg gccgtcgttt tacggtacca
     4201 tcgatgatga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     4261 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4321 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4381 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     4441 attatgatcc tctagatcag atctgcgaag atacggccac gggtgctctt gatcctgtgg
     4501 ctgattttgg actgtgctgc tcgcagctgc tgatgaatca catacttcct ccattttctt
     4561 ccactgattg actgttataa tttccctaat ttccaggtca aggtgctgtg cattgtggta
     4621 atagatgtga catgacgtca cttccaaagg accaatgaac atgtctgacc aatttcatat
     4681 aatgtgaaaa cgattttcat aggcagaata aataacattt aaattaaact gggcatcagc
     4741 gcaattcaat tggtttggta atagcaaggg aaaatagaat gaagtgatct ccaaaaaata
     4801 agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa
     4861 atgtaattaa gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc
     4921 cccaaaaata atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct
     4981 ggttcttgac cccctacctt cagcaagccc agcagatcca ctagttctag agcggccgcc
     5041 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     5101 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5161 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5221 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5281 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5341 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5401 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5461 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5521 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5581 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5641 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5701 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5761 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5821 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5881 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5941 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6001 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6061 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6121 ggggtctgac gctcagtgga acgaaaactc acgttaaggc attttggtca tgagattatc
     6181 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6241 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6301 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6361 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6421 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6481 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6541 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6601 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6661 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6721 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6781 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6841 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6901 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6961 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7021 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7081 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7141 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7201 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     7261 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     7321 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     7381 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     7441 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac gctggctaag aactcatcag
     7501 cctccccggt ccatctaccc acgtaccaat gcaccaattg gccacaatga cggctactac
     7561 atggtgccat tccttcctct ttataggaat ggagactacc tcctgtccaa caatgctctt
     7621 ggatacgagt acgcctacct gttggaccca ggtcattgca caacaccaga aatgccctct
     7681 gatctgcaaa agacgtgaat atctgttcag acacccatat ccactctgtt ccacacaggt
     7741 cagaggtttg tccaggagtt cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa
     7801 gtgaaagtac aagtacttag ggaaaatttt actcaattaa aagtaaaagt atctggctag
     7861 aatcttactt gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga gtattaagga
     7921 agtaaaagta aaagcaagaa agaaaactag agattcttgt ttaagctttt aatctcaaaa
     7981 aacattaaat gaaatgcata caaggtttta tcctgcttta gaactgtttg tatttaatta
     8041 tcaaactata agacagacaa tctaatgcca gtacacgcta ctcaaagttg taaaacctca
     8101 gatttaactt cagtagaagc tgattctcaa aattgttagt gtcaagccta gctcttttgg
     8161 ggctgaaaag caatcctgca gtgctgaaaa gcctctcaca ggcagccgat gcgggaagag
     8221 gtgtattagt cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga gactccctgg
     8281 tgtctgaaac acaggccaga tgggccctcg agcaggaaac agctatgacc atgattacgc
     8341 caagctatca actttgtata gaaaagttgg ctccgaattc gcccttaaaa ctcgagacca
     8401 gcaaagttct agaatttgtc gaaacattta tgttatatat ttcctgaaaa aaattctgag
     8461 taagttctta agtgttattg ccagcaacat aaacaacaga cggcaaaatg aataaatgat
     8521 aacaaagcag taggcttaaa taaacctaat ttttataggc tgttctctac aaccctcaaa
     8581 cagtgattag ttttgtactt ataaacttgc cctttcattc atatttcaag aaaattggtt
     8641 cagaagatct ggatattcta gcagttgttc aagctcatgg agggatcagt gacctgattc
     8701 cacaatgact aggcctaatc cagaaattag atgactgtca acataaaaag gcacagcact
     8761 cactagctgc cctatatatt ttattatatt ttacatatat tattttattt atttagctct
     8821 gagtgctgta ctttctggtt aaagaaaact gcttacaaca gctaacctgt actacctcag
     8881 gctcagggaa tttggaacag gtttgtctgg tttgtttctt taaccatgca tgcttgtttt
     8941 caactatggc aacacagtca catgggacat tacagaaatg atttgtcgat gacatgcgac
     9001 ttttctttaa taaagcgcaa agatcccaaa aagcaaactt ttaacaaaaa tcatataatt
     9061 atattttcaa tccagctttg tagcaacttt gtgctgctgt tcactcagca acagatagtc
     9121 agtataaggt cagtgtgtct caaagcagtg ccatctgttt cacacattgc gttctatata
     9181 taagtgtgct ggttgacacg acactgtata aggcctaggc taaaacacaa acaatgtaga
     9241 atgacactgt gttttttttg taaacaaatg ttgtttttgg ttaaacatct ttg
//
