LOCUS       sequence-274365-        7607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  chemigenetic voltage indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7607)
  AUTHORS   Deo C, Abdelfattah AS, Bhargava HK, Berro AJ, Falco N, Farrants H, 
            Moeyaert B, Chupanova M, Lavis LD, Schreiter ER
  TITLE     The HaloTag as a general scaffold for far-red tunable chemigenetic 
            indicators.
  JOURNAL   Nat Chem Biol. 2021 Jun;17(6):718-723. doi: 
            10.1038/s41589-021-00775-w. Epub 2021 Apr 1.
  PUBMED    33795886
REFERENCE   2  (bases 1 to 7607)
  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: 
            "10.1038/s41589-021-00775-w"; journalName: "Nat Chem Biol"; date: 
            "2021-06"; volume: "17"; issue: "6"; pages: "718-723"
COMMENT     Sequence Label: pCAG-HASAP
FEATURES             Location/Qualifiers
     source          1..7607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..654
                     /label=chicken beta-actin promoter
     intron          655..1671
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1595..1617
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1679..1698
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1831..1840
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3463..3480)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3469..3693
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3739..4167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3826..3845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4036..4057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4176..4196)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4181..4510
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4361..4496
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4423..4442
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4521..5120
                     /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(4521..4540)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5017..5037
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5280..5401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5317..5336)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5371..5390
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5450..5466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5450..5466)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5463..5485)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5474..5490
                     /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(5498..5504,5505..5522,5523..5528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5543..5564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5681..5698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5852..6440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5932..5951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6611..7402,7403..7471))
                     /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     7234..7253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7472..7576)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
      541 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      601 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
      661 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
      721 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      781 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
      841 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
      901 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
      961 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     1081 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     1321 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     1381 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     1441 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     1561 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1621 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1681 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga ggaattcact
     1741 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     1801 aaataccact gagatctgca tgagctagcc gccaccatgg agacgactgt gaggtatgaa
     1861 caggggtcag agctcactaa aacttcgagc tctccaacag cagatgagcc cacgataaag
     1921 attgatgatg gtcgtgatga gggtaatgaa caagacagct gttccaatac cattaggaga
     1981 aaaatttccc cgtttgtgat gtcatttgga ttcagagtat ttggagttgt gcttatcatt
     2041 gtagacatca tagtggtgat tgtggatctg gccatcagtg agaagaaaag aggcattaga
     2101 gagattcttg aaggtgtttc cctggctata gcactcttct tccttgttga tgttctcatg
     2161 agagtgtttg ttgaaggctt caagaactat ttccggtcca aactgaatac tttggatgca
     2221 gtcatagtag tgggcactct gctaattaat atgacctact ccttctctga ccttgctgcc
     2281 tttgcccgcg agaccttcca ggccttccgc accaccgacg tcggccgcaa gctgatcatc
     2341 gatcagaacg tttttatcga gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa
     2401 gtcgagatgg accattaccg cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg
     2461 cgcttcccaa acgagctgcc aatcgccggt gagccagcga acatcgtcgc gctggtcgaa
     2521 gaatacatgg actggctgca ccagtcccct gtcccgaagc tgctgttctg gggcacccca
     2581 ggcgttctga tcccaccggc cgaagccgct cgcctggcca aaagcctgcc taactgcaag
     2641 gctgtggaca tcggcccggg tctgaatctg ctgcaagaag acaacccgga cctgatcggc
     2701 agcgagatcg cgcgctggct gtcgacgctc gagatttccg gcgagccaac cactggaggc
     2761 agcggaggca caggaggcag cggaggcaca ggaggcagca tggcagaaat cggtactggc
     2821 tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt
     2881 ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg
     2941 tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc
     3001 ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc
     3061 atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg
     3121 ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca
     3181 tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgccgggaca
     3241 gatcagatgc ctcaaatggt gacacttttg cgagttctgc gaatagtgat cctgattcga
     3301 atctttcgcc ttgccagcca gaagaaacaa ctggaggtag taacataatg aaagcttgct
     3361 accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca gcacagtggc
     3421 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     3481 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3541 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3601 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3661 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3721 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3781 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3841 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3901 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3961 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4021 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4081 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4141 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4201 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4261 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4321 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4381 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4441 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4501 cttttgcaaa aagctcccgc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     4561 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     4621 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     4681 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     4741 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     4801 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     4861 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     4921 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     4981 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5041 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5101 cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5161 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5221 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5401 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5461 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5521 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5581 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5641 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5701 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5761 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5821 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5881 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5941 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6001 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6061 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6121 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6181 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6241 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6301 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6361 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6421 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6481 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6541 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6601 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6661 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6721 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6781 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6841 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6901 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6961 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7021 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7081 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7141 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7201 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7261 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7321 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7381 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7441 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7501 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7561 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
