LOCUS       sequence-254366-        8002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express soma-localized QuaAr3_Q95D_H106N_E214V-HaloTag in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8002)
  AUTHORS   Abdelfattah AS, Valenti R, Zheng J, Wong A, Podgorski K, Koyama M, 
            Kim DS, Schreiter ER
  TITLE     A general approach to engineer positive-going eFRET voltage 
            indicators.
  JOURNAL   Nat Commun. 2020 Jul 10;11(1):3444. doi: 10.1038/s41467-020-17322-1.
  PUBMED    32651384
REFERENCE   2  (bases 1 to 8002)
  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 
            Commun."; date: "2020-07-10"; volume: "11(1):3444. doi"; pages: " 
            10.1038/s41467-020-17322-1"
COMMENT     Sequence Label: pCAG-QuaAr3_Q95D_H106N_E214V-HaloTag-ST
FEATURES             Location/Qualifiers
     source          1..8002
                     /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(2033..2050)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2039..2263
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2309..2737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2396..2415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2606..2627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2746..2766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2751..3080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2931..3066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2993..3012
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3091..3690
                     /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(3091..3110)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3587..3607
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3850..3971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3887..3906)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3941..3960
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4020..4036)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4020..4036)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4033..4055)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4044..4060
                     /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(4068..4074,4075..4092,4093..4098))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4113..4134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4251..4268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4422..5010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4502..4521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5181..5972,5973..6041))
                     /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     5804..5823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6042..6146)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6178..6557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6834..7850
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7774..7796
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7858..7877
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gctagccgcc accatggtat ctatcgcact tcaagccggt tatgatctcc tgggagatgg
       61 gcggcctgaa acactttggc tcgggatcgg cactttgctt atgctcattg gcacgtttta
      121 ttttctggta cgaggatggg gggtcactga caaagatgca agggagtact acgcagtgac
      181 gattcttgta agtggcatcg ccagtgctgc ttacctgtca atgtttttcg gaatcggcct
      241 cacagaggta agcgttggtg gggaaatgct cgacatttat tatgctaggt acgccgactg
      301 gttgtttacg acaccactcc tgctgctcaa cttggccttg ttggctaagg tagatagggt
      361 cactatcggc accctggtag gagtggatgc actgatgatt gtaacggggc tgattggagc
      421 cttgagtcat actgcaatcg caaggtactc atggtggctg ttctctacaa tatgtatgat
      481 tgtggtgctc tatgtgttgg ctacgtcact tcggtccgct gcacgcgaaa gaggacctga
      541 agtcgccagt acttttaata ccctgacagc tttggtgctc gtactgtgga ccgcatatcc
      601 cattctttgg attattggta cggagggagc aggtgttgtt ggactcggta tcgtgaccct
      661 cctcttcatg gttctggacg taactgcaaa ggttgggttt ggcttcatcc ttctcaggag
      721 tcgcgcaata ttgggagaca ctgagatcgg taccggcttt ccattcgacc cccattatgt
      781 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt
      841 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt
      901 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc
      961 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct
     1021 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg
     1081 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccaccctat
     1141 cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac
     1201 cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat
     1261 gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa
     1321 tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc
     1381 agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc
     1441 gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct
     1501 ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca
     1561 agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcga cgctcgagat
     1621 ttccggcgag ccaaccacta agagcaggat caccagcgag ggcgagtaca tccccctgga
     1681 ccagatcgac atcaacgtgt tctgctacga gaacgaggtg caaagtcagc ctatcctgaa
     1741 cacaaaggaa atggctccac agtctaagcc tcccgaagag cttgagatgt ccagtatgcc
     1801 aagtcccgtg gctcccctcc ctgccaggac tgaaggagtg attgacatga ggagtatgtc
     1861 atctattgat agcttcatct cttgcgcaac agatttcccc gaggctactc gattctaatg
     1921 aaagcttgct accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca
     1981 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2041 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2101 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2161 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2221 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2281 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2341 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2401 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2461 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2521 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2581 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2641 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2701 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2761 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2821 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2881 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3001 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3061 gaggcctagg cttttgcaaa aagctcccgc atgaccgagt acaagcccac ggtgcgcctc
     3121 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     3181 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     3241 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     3301 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     3361 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     3421 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     3481 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     3541 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     3601 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     3661 tggtgcatga cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc gacgcccaac
     3721 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3781 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3841 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3901 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3961 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4021 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4081 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4141 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4201 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4261 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4321 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4861 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4981 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5041 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5101 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5161 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5221 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5281 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5341 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5401 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5461 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5521 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5581 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5641 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5701 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5761 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5821 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5881 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5941 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6001 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6061 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6121 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     6241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     6301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     6361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     6421 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     6481 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     6541 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     6601 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     6661 gggcgggggg ggggggnggs gcgccrggsg gggsggggsg gggsgrgggg sggggsgggg
     6721 sgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
     6781 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
     6841 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
     6901 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     6961 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
     7021 aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg
     7081 tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc
     7141 gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg
     7201 ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     7261 gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc
     7321 cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg
     7381 ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc
     7441 ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc
     7501 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     7561 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     7621 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     7681 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     7741 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     7801 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     7861 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgag gaattcactc
     7921 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     7981 aataccactg agatctgcat ga
//
