LOCUS       sequence-225067-        7764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Voltron in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7764)
  AUTHORS   Abdelfattah AS, Kawashima T, Singh A, Novak O, Liu H, Shuai Y, Huang
            YC, Campagnola L, Seeman SC, Yu J, Zheng J, Grimm JB, Patel R, 
            Friedrich J, Mensh BD, Paninski L, Macklin JJ, Murphy GJ, Podgorski 
            K, Lin BJ, Chen TW, Turner GC, Liu Z, Koyama M, Svoboda K, Ahrens 
            MB, Lavis LD, Schreiter ER
  TITLE     Bright and photostable chemigenetic indicators for extended in vivo 
            voltage imaging.
  JOURNAL   Science. 2019 Aug 1. pii: science.aav6416. doi: 
            10.1126/science.aav6416.
  PUBMED    31371562
REFERENCE   2  (bases 1 to 7764)
  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: "Science. 
            2019 Aug 1. pii: science.aav6416. doi: 10.1126/science.aav6416."
COMMENT     Sequence Label: pCAG-Voltron
FEATURES             Location/Qualifiers
     source          1..7764
                     /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(1787..1804)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1793..2017
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2063..2491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2150..2169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2360..2381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2500..2520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2505..2834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2685..2820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2747..2766
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2843..3442
                     /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(2843..2862)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3339..3359
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3602..3723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3639..3658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3693..3712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3772..3788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3772..3788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3785..3807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3796..3812
                     /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(3820..3826,3827..3844,3845..3850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3865..3886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4003..4020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4174..4759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4254..4273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4930..5721,5722..5790))
                     /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     5553..5572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5791..5895)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5941..6320
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6322..6595
                     /label=chicken beta-actin promoter
     intron          6596..7612
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7536..7558
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7620..7639
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gctagccgcc accatggctg acgtggaaac cgagaccggc atgattgcac agtggattgt
       61 ctttgctatt atggctgctg ctgctattgc ttttggagtg gctgtgcact ttcggccttc
      121 agagctgaag agcgcatact atatcaacat tgccatctgc actatcgccg ctaccgctta
      181 ctatgcaatg gccgtgaact accaggacct gacaatgaat ggtgaaaggc aggtggtcta
      241 cgcaagatat attaactggg tgctgaccac accactgctc ctgctcgatc tcatcgtcat
      301 gaccaagatg ggcggagtga tgatttcttg ggtcatcggc gcagacattt tcatgatcgt
      361 gtttggtatt ctgggcgcct tcgaggatga acacaagttc aaatgggtgt actttatcgc
      421 tggatgtgtg atgcaggcag tcctgacata cgggatgtat aacgccactt ggaaagacga
      481 tctgaagaaa agccccgagt accatagctc ctatgtcagt ctgctcgtct tcctgtcaat
      541 cctctgggtg ttttatcctg tcgtgtgggc tttcgggtct ggtagtggcg tgctgtccgt
      601 cgacaatgag gccattctca tgggaatcct ggatgtgctc gctaagccac tgtttggaat
      661 ggggtgcctc attgcccatg agactatctt caagatcggt actggctttc cattcgaccc
      721 ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg
      781 cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat
      841 cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc
      901 cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat
      961 cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg
     1021 tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat
     1081 ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt
     1141 ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac
     1201 gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc
     1261 gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc
     1321 cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc
     1381 ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc
     1441 cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa
     1501 tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtcgac
     1561 gctcgagatt tccggcgagc caaccactaa gagcaggatc accagcgagg gcgagtacat
     1621 ccccctggac cagatcgaca tcaacgtgtt ctgctacgag aacgaggtgt aatgaaagct
     1681 tggtaccgag ctcggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag
     1741 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     1801 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1861 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1921 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1981 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2041 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2101 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2161 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2221 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2281 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2341 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2401 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2461 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2521 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2581 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2641 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2701 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2761 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2821 taggcttttg caaaaagctc ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     2881 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     2941 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3001 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3061 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3121 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3181 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3241 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     3301 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     3361 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     3421 gacccgcaag cccggtgcct gattcgaaat gaccgaccaa gcgacgccca acctgccatc
     3481 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3541 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3601 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3661 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3721 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3781 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3841 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3901 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3961 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4681 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     4741 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4801 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4861 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4921 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4981 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5041 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5101 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5161 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5221 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5281 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5341 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5401 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5461 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5521 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5581 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5641 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5701 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5761 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5821 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5881 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcggggtc
     5941 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6001 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6061 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6121 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6181 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6241 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     6301 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     6361 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     6421 tgggggcggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
     6481 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     6541 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
     6601 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
     6661 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
     6721 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
     6781 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg
     6841 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
     6901 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
     6961 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     7021 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
     7081 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
     7141 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     7201 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
     7261 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     7321 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     7381 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     7441 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     7501 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     7561 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     7621 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcctcg aggaattcac
     7681 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     7741 caaataccac tgagatctgc atga
//
