LOCUS       sequence-267119-        7617 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 7617)
  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 7617)
  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..7617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1627..1644)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1633..1857
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1903..2331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1990..2009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2200..2221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2340..2360)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2345..2674
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2525..2660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2587..2606
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2683..3282
                     /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(2683..2702)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3179..3199
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3442..3563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3479..3498)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3533..3552
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3612..3628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3612..3628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3625..3647)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3636..3652
                     /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(3660..3666,3667..3684,3685..3690))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3705..3726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3843..3860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4014..4599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4094..4113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4770..5561,5562..5630))
                     /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     5393..5412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5631..5735)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5781..6160
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6162..6435
                     /label=chicken beta-actin promoter
     intron          6436..7452
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7376..7398
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7460..7479
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      join(7612..7617,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggagacga ctgtgaggta tgaacagggg tcagagctca ctaaaacttc gagctctcca
       61 acagcagatg agcccacgat aaagattgat gatggtcgtg atgagggtaa tgaacaagac
      121 agctgttcca ataccattag gagaaaaatt tccccgtttg tgatgtcatt tggattcaga
      181 gtatttggag ttgtgcttat cattgtagac atcatagtgg tgattgtgga tctggccatc
      241 agtgagaaga aaagaggcat tagagagatt cttgaaggtg tttccctggc tatagcactc
      301 ttcttccttg ttgatgttct catgagagtg tttgttgaag gcttcaagaa ctatttccgg
      361 tccaaactga atactttgga tgcagtcata gtagtgggca ctctgctaat taatatgacc
      421 tactccttct ctgaccttgc tgcctttgcc cgcgagacct tccaggcctt ccgcaccacc
      481 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
      541 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
      601 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
      661 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
      721 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
      781 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
      841 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt
      901 tccggcgagc caaccactgg aggcagcgga ggcacaggag gcagcggagg cacaggaggc
      961 agcatggcag aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc
     1021 gagcgcatgc actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac
     1081 ggtaacccga cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat
     1141 cgctgcattg ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat
     1201 ttcttcgacg accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag
     1261 gtcgtcctgg tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat
     1321 ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac
     1381 gaatggccag aatttgccgg gacagatcag atgcctcaaa tggtgacact tttgcgagtt
     1441 ctgcgaatag tgatcctgat tcgaatcttt cgccttgcca gccagaagaa acaactggag
     1501 gtagtaacat aatgaaagct tgctaccgag ctcggatcca ctagtccagt gtggtggaat
     1561 tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct
     1621 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1681 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1741 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1801 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     1861 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     1921 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1981 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2041 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2101 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2161 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2221 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2281 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2341 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2401 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2461 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2521 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2581 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2641 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccatgaccga gtacaagccc
     2701 acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg
     2761 ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc
     2821 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     2881 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     2941 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3001 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3061 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3121 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3181 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     3241 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gattcgaaat gaccgaccaa
     3301 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3361 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3421 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3481 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3541 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     3601 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3661 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3721 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt
     4561 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4621 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4681 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4741 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4801 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4861 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4921 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4981 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5041 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5101 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5161 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5221 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5281 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5341 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5401 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5461 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5521 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5581 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5641 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5701 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5761 gacgtcgacg gatcggggtc gacattgatt attgactagt tattaatagt aatcaattac
     5821 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     5881 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     5941 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6001 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6061 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     6121 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
     6181 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
     6241 ttaattattt tgtgcagcga tgggggcggg gggggggggc gcgcgccagg cggggcgggg
     6301 cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
     6361 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
     6421 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
     6481 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
     6541 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
     6601 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
     6661 tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc
     6721 ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag
     6781 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc
     6841 tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg
     6901 caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
     6961 ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg
     7021 gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc
     7081 ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa
     7141 tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg
     7201 aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag
     7261 gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag
     7321 cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc
     7381 ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc
     7441 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
     7501 gaattcctcg aggaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     7561 gtggccaatg ccctggctca caaataccac tgagatctgc atgagctagc cgccacc
//
