LOCUS       sequence-240797-        7377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for voltage imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7377)
  AUTHORS   Yang HH, St-Pierre F, Sun X, Ding X, Lin MZ, Clandinin TR
  TITLE     Subcellular Imaging of Voltage and Calcium Signals Reveals Neural 
            Processing In Vivo.
  JOURNAL   Cell. 2016 Jun 30;166(1):245-57. doi: 10.1016/j.cell.2016.05.031. 
            Epub 2016 Jun 2.
  PUBMED    27264607
REFERENCE   2  (bases 1 to 7377)
  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"; doi: 
            "10.1016/j.cell.2016.05.031"; journalName: "Cell"; date: 
            "2016-06-30- 30"; volume: "166"; issue: "1"; pages: "245-57"
COMMENT     Sequence Label: ASAP2f
FEATURES             Location/Qualifiers
     source          1..7377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          168..1184
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1108..1130
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1192..1211
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1344..1353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2745..2762)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2751..2975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3021..3449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3108..3127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3318..3339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3458..3478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3463..3792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3643..3778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3705..3724
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3803..4402
                     /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(3803..3822)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4299..4319
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4562..4683
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4599..4618)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4653..4672
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4732..4748)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4732..4748)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4745..4767)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4756..4772
                     /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(4780..4786,4787..4804,4805..4810))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4825..4846
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4963..4980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5134..5722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5214..5233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5893..6684,6685..6753))
                     /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     6516..6535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6754..6858)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6890..7269
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggggggggg gggsgcgccr ggsggggggg gggggggggg gggggggggg ggggsgrggs
       61 ggrgrggkgs ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
      121 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
      181 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
      241 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
      301 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
      361 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
      421 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
      481 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
      541 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
      601 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
      661 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
      721 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
      781 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
      841 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
      901 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
      961 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     1021 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     1081 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1141 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1201 tggttattgt gctgtctcat cattttggca aagaattcct cgaggaattc actcctcagg
     1261 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     1321 actgagatct gcatgagcta gccgccacca tggagacgac tgtgaggtat gaacaggggt
     1381 cagagctcac taaaacttcg agctctccaa cagcagatga gcccacgata aagattgatg
     1441 atggtcgtga tgagggtaat gaacaagaca gctgttccaa taccattagg agaaaaattt
     1501 ccccgtttgt gatgtcattt ggattcagag tatttggagt tgtgcttatc attgtagaca
     1561 tcatagtggt gattgtggat ctggccatca gtgagaagaa aagaggcatt agagagattc
     1621 ttgaaggtgt ttccctggct atagcactct tcttccttgt tgatgttctc atgagagtgt
     1681 ttgttgaagg cttcaagaac tatttccggt ccaaactgaa tactttggat gcagtcatag
     1741 tagtgggcac tctgctaatt aatatgacct actccttctc tgaccttagc tttaacagcc
     1801 ataacgtgta tattaccgcg gataaacaga aaaacggcat taaagcgaac tttaccgtgc
     1861 gccataacgt ggaagatggc agcgtgcagc tggcggatca ttatcagcag aacaccccga
     1921 ttggcgatgg cccggtgctg ctgccggata accattatct gagcacccag accgtgctga
     1981 gcaaagatcc gaacgaaaaa cgcgatcaca tggtgctgct ggaatttgtg accgcagcgg
     2041 gcattacaca cggcatggat gaactgtatg gcggcaccgg cggcagcgcg agccagggcg
     2101 aagaactgtt taccggcgtg gtgccgattc tggtggaact ggatggcgat gtgaacggcc
     2161 ataaatttag cgtgcgcggc gaaggcgaag gcgatgcgac cattggcaaa ctgaccctga
     2221 aatttatttg caccaccggc aaactaccgg tgccgtggcc gaccctggtg accaccttaa
     2281 cctatggcgt gcagtgcttt agccgctatc cggatcatat gaaacgccat gattttttta
     2341 aaagcgcgat gccggaaggc tatgtgcagg aacgcaccat tagctttaaa gatgatggca
     2401 aatataaaac ccgcgcggtg gtgaaatttg aaggcgatac cctggtgaac cgcattgaac
     2461 tgaaaggcac cgattttaaa gaagatggca acattctggg gcataaactg gaatataaca
     2521 cagatcagat gccgcagatg gttactcttc ttcgagttct gagaattgtt atcttaataa
     2581 gaatatttcg cctggcttca cagaagaaac aacttgaagt ggtaacctaa tgaaagcttg
     2641 gtaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc cagcacagtg
     2701 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2761 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2821 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2881 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2941 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3001 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3061 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3121 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3181 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3241 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3301 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3361 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3421 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3541 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3601 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3661 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3721 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3781 ggcttttgca aaaagctccc gcatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3841 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3901 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3961 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4021 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4081 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4141 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4201 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4261 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4321 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4381 gacccgcaag cccggtgcct gattcgaaat gaccgaccaa gcgacgccca acctgccatc
     4441 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4501 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4561 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4621 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4681 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4741 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4801 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4861 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4921 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4981 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5041 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5101 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5161 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5221 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5281 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5341 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5401 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5461 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5521 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5581 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5641 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5701 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5761 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5821 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5881 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5941 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6001 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6061 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6121 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6181 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6241 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6301 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6361 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6421 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6481 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6541 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6601 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6661 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6721 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6781 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6841 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acattgatta
     6901 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     6961 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     7021 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     7081 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     7141 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     7201 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     7261 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     7321 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcg
//
