LOCUS       sequence-274366-        6412 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 6412)
  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 6412)
  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-HaloCaMP1a-EGFP
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        148..595
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             692..709
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1736..2176
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     misc_feature    2935..3523
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2988..3008)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3406..3417)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3526..3542)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3527..3543)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3572..3693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3583..3602)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3637..3656
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4072..4660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4152..4171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4831..5622,5623..5691))
                     /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     5454..5473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5692..5796)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5822..6277)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5959..5980)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6171..6190
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(6349..6412,1..66)
                     /label=AAV2 ITR
ORIGIN
        1 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
       61 gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga
      121 agatctctgc agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg
      181 gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat
      241 tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc
      301 gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg
      361 ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc
      421 ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc
      481 gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc
      541 gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgaa
      601 ttcaagctgc tagcaaggat ccgccaccat gctgcagaac gagcttgctc ttaagttggc
      661 tggacttgat attaacaaga ctggaggttc tcatcatcac caccaccatg gatccgcccg
      721 tcgtaaatgg cagaaaacag gccatgcggt tcgtgctatc ggtcgtttgt cgagccctga
      781 gtttgcagcg gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat
      841 cgatcagaac gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga
      901 agtcgagatg gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg
      961 gcgcttccca aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga
     1021 agaatacatg gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc
     1081 aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa
     1141 ggctgtggac atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg
     1201 cagcgagatc gcgcgctggc tgtcgacgct cgagatttcc ggcggaggaa caggtggttc
     1261 tggtggaaca gggggtagcg gaggtacagg aggaagtatg gcggagatcg gaactggatt
     1321 cccgtttgat ccgcattatg tggaagttct gggagagcgc atgcattatg tggacgttgg
     1381 tcctcgtgat gggacaccag tgctgttcct tcacggcaat ccgacatcgt cgtacgtgtg
     1441 gcgtaatatc atcccgcacg ttgcccccac gcaccgctgc attgcccctg acttaattgg
     1501 tatggggaaa agtgataagc ctgatctggg gtacttcttt gacgaccacg tacgcttcat
     1561 ggatgctttt attgaagcat tgggtttgga ggaagtagtt ttggtgatcc atgattgggg
     1621 tagtgctctg gggttccatt gggccaagcg taacccagaa cgcgtgaaag gaattgcctt
     1681 tatggagttc atccgtccga ttccaacatg ggacgaatgg cgagaatttg cacgcgatca
     1741 attaacagag gaacagattg cggagtttaa ggaagcgttc tctttatttg ataaggatgg
     1801 cgacggtaca atcactacta aagaattggg aacagtcatg cgctcattgg ggcaaaatcc
     1861 gacagaggct gaattgcagg acatgattaa cgaggtagac gccgatggga acgggactat
     1921 cgactttccg gaatttctta ctatgatggc acgcaaaatg aaagataccg attctgaaga
     1981 agagatccgt gaagctttcc gtgtttttga taaagatggg aacggctaca tcagtgctgc
     2041 tgagttacgc catgtgatga caaatctggg ggaaaaactt accgacgaag aagtagacga
     2101 aatgattcgc gaggcggata ttgacgggga tggacaagta aactacgagg aatttgtgca
     2161 gatgatgacc gccaagctcg agatttccgg cggcggaagc ggcatggtct ctaaggggga
     2221 ggaactcttt accggtgtgg tgccaatact tgtagagctc gatggggacg ttaatggaca
     2281 caaattttct gtctcaggtg agggtgaggg agatgcaact tatggaaaac tgacattgaa
     2341 attcatctgc acaacaggca aattgcctgt cccctggcct accctggtaa ccaccctcac
     2401 ttatggcgtt cagtgcttct cccggtatcc tgatcacatg aaacagcatg atttcttcaa
     2461 atcagcaatg cccgaaggtt atgttcaaga gcggaccatc tttttcaagg acgatggaaa
     2521 ctataaaacc agagctgagg ttaagttcga gggagatact ttggtgaatc ggatcgaact
     2581 gaaggggata gacttcaaag aggatgggaa cattttgggg cataaactcg aatataacta
     2641 caactcccac aacgtgtata tcatggctga caaacagaag aacgggatca aagtaaattt
     2701 taagatacga cacaatatag aggacggtag tgtccaactt gctgatcact accagcagaa
     2761 tacacccatc ggagacggac ccgttctctt gcccgataat cactatctct ctacacaatc
     2821 cgctctgtct aaagatccaa acgaaaagcg ggaccacatg gtactgcttg agttcgttac
     2881 agccgctggt atcaccttgg gcatggatga gttgtaaaat aaaagcttat cgataatcaa
     2941 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3001 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3061 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3121 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3181 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3241 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3301 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3361 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3421 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3481 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     3541 cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3601 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3661 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3721 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3781 ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     3841 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3901 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3961 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4021 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4081 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4141 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4201 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4261 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4321 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4381 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4441 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4501 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4561 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4681 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4741 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4801 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4861 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4921 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4981 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5041 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5101 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5161 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5221 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5281 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5341 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5401 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5461 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5521 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5581 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5641 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5701 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5761 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5821 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     5881 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     5941 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     6001 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     6061 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     6121 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     6181 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     6241 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     6301 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc
     6361 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tc
//
