LOCUS       sequence-267142-        6562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  chemigenetic calcium indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6562)
  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 6562)
  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: pAAV-synapsin-HaloCaMP1b-EGFP
FEATURES             Location/Qualifiers
     source          1..6562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     promoter        267..714
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             811..828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1861..2301
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(2323..2325,2326..2328,2329..3039)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3055..3643
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3108..3128)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3526..3537)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3646..3662)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3647..3663)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3692..3813
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3703..3722)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3757..3776
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3975..4104
                     /label=AAV2 ITR
     rep_origin      4176..4631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4263..4282)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4473..4494
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4657..4761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4762..4830,4831..5622)
                     /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     complement(4980..4999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5793..6381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6282..6301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatctctgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
      301 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
      361 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
      421 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
      481 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
      541 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
      601 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
      661 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat
      721 tcaagctgct agcaaggatc cgccaccatg ctgcagaacg agcttgctct taagttggct
      781 ggacttgata ttaacaagac tggaggttct catcatcacc accaccatgg agtgcgggtt
      841 attcccagac ttgataccct gatactcgtg aaagcaatgg gccaccgaaa acgattcggt
      901 aaccccttta ggcctaagga gaccttccag gccttccgca ccaccgacgt cggccgcaag
      961 ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg
     1021 ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag
     1081 ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg
     1141 ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg
     1201 ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct
     1261 aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac
     1321 ctgatcggca gcgagatcgc gcgctggctg tcgacgctcg agatttccgg cggaggaaca
     1381 ggtggttctg gtggaacagg gggtagcgga ggtacaggag gaagtatggc ggagatcgga
     1441 actggattcc cgtttgatcc gcattatgtg gaagttctgg gagagcgcat gcattatgtg
     1501 gacgttggtc ctcgtgatgg gacaccagtg ctgttccttc acggcaatcc gacatcgtcg
     1561 tacgtgtggc gtaatatcat cccgcacgtt gcccccaagc accgctgcat tgcccctgac
     1621 ttaattggta tggggaaaag tgataagcct gatctggggt acttctttga cgaccacgta
     1681 cgcttcatgg atgcttttat tgaagcattg ggtttggagg aagtagtttt ggtgatccat
     1741 gattggggta gtgctctggg gttccattgg gccaagcgta acccagaacg cgtgaaagga
     1801 attgccttta tggagttcat ccgtccgatt ccaacatggg acgaatggcc ttttgcacgc
     1861 gatcaattaa cagaggaaca gattgcggag tttaaggaag cgttctcttt atttgataag
     1921 gatggcgacg gtacaatcac tactaaagaa ttgggaacag tcatgcgctc attggggcaa
     1981 aatccgacag aggctgaatt gcaggacatg attaacgagg tagacgccga tgggaacggg
     2041 actatcgact ttccggaatt tcttactatg atggcacgca aaatgaaaga taccgattct
     2101 gaagaagaga tccgtgaagc tttccgtgtt tttgataaag atgggaacgg ctacatcagt
     2161 gctgctgagt tacgccatgt gatgacaaat ctgggggaaa aacttaccga cgaagaagta
     2221 gacgaaatga ttcgcgaggc ggatattgac ggggatggac aagtaaacta cgaggaattt
     2281 gtgcagatga tgaccgccaa gatttccggc ggcggaagcg gcatggtctc taagggggag
     2341 gaactcttta ccggtgtggt gccaatactt gtagagctcg atggggacgt taatggacac
     2401 aaattttctg tctcaggtga gggtgaggga gatgcaactt atggaaaact gacattgaaa
     2461 ttcatctgca caacaggcaa attgcctgtc ccctggccta ccctggtaac caccctcact
     2521 tatggcgttc agtgcttctc ccggtatcct gatcacatga aacagcatga tttcttcaaa
     2581 tcagcaatgc ccgaaggtta tgttcaagag cggaccatct ttttcaagga cgatggaaac
     2641 tataaaacca gagctgaggt taagttcgag ggagatactt tggtgaatcg gatcgaactg
     2701 aaggggatag acttcaaaga ggatgggaac attttggggc ataaactcga atataactac
     2761 aactcccaca acgtgtatat catggctgac aaacagaaga acgggatcaa agtaaatttt
     2821 aagatacgac acaatataga ggacggtagt gtccaacttg ctgatcacta ccagcagaat
     2881 acacccatcg gagacggacc cgttctcttg cccgataatc actatctctc tacacaatcc
     2941 gctctgtcta aagatccaaa cgaaaagcgg gaccacatgg tactgcttga gttcgttaca
     3001 gccgctggta tcaccttggg catggatgag ttgtataaat aaaagcttat cgataatcaa
     3061 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3121 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3181 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3241 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3301 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3361 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3421 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3481 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3541 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3601 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     3661 cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3721 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3781 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3841 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3901 ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     3961 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4021 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     4081 ctcagtgagc gagcgagcgc gcagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4141 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     4201 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4261 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4321 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4381 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4441 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4501 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4561 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4621 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4741 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4801 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4861 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4921 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4981 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5041 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5101 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5161 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5221 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5281 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5341 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5401 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5461 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5521 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5581 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5641 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5701 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5761 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5821 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5881 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5941 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6001 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6061 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6121 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6181 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6241 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6301 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6361 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6421 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6481 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6541 tcagtgagcg aggaagcgga ag
//
