LOCUS       sequence-275495-        6500 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 6500)
  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 6500)
  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..6500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        212..659
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             756..773
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1806..2246
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(2268..2270,2271..2273,2274..2984)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3000..3588
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3053..3073)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3471..3482)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3591..3607)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3592..3608)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3637..3758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3648..3667)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3702..3721
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3920..4049
                     /label=AAV2 ITR
     rep_origin      4121..4576
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4208..4227)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4418..4439
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4602..4706
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4707..4775,4776..5567)
                     /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(4925..4944)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5738..6326
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6227..6246
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg accaggatga
      241 ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc
      301 ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag
      361 gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg
      421 cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa
      481 ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc
      541 acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact
      601 cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt
      661 cgaattcaag ctgctagcaa ggatccgcca ccatgctgca gaacgagctt gctcttaagt
      721 tggctggact tgatattaac aagactggag gttctcatca tcaccaccac catggagtgc
      781 gggttattcc cagacttgat accctgatac tcgtgaaagc aatgggccac cgaaaacgat
      841 tcggtaaccc ctttaggcct aaggagacct tccaggcctt ccgcaccacc gacgtcggcc
      901 gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc
      961 gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc
     1021 gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg
     1081 tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt
     1141 tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc
     1201 tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc
     1261 cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt tccggcggag
     1321 gaacaggtgg ttctggtgga acagggggta gcggaggtac aggaggaagt atggcggaga
     1381 tcggaactgg attcccgttt gatccgcatt atgtggaagt tctgggagag cgcatgcatt
     1441 atgtggacgt tggtcctcgt gatgggacac cagtgctgtt ccttcacggc aatccgacat
     1501 cgtcgtacgt gtggcgtaat atcatcccgc acgttgcccc caagcaccgc tgcattgccc
     1561 ctgacttaat tggtatgggg aaaagtgata agcctgatct ggggtacttc tttgacgacc
     1621 acgtacgctt catggatgct tttattgaag cattgggttt ggaggaagta gttttggtga
     1681 tccatgattg gggtagtgct ctggggttcc attgggccaa gcgtaaccca gaacgcgtga
     1741 aaggaattgc ctttatggag ttcatccgtc cgattccaac atgggacgaa tggccttttg
     1801 cacgcgatca attaacagag gaacagattg cggagtttaa ggaagcgttc tctttatttg
     1861 ataaggatgg cgacggtaca atcactacta aagaattggg aacagtcatg cgctcattgg
     1921 ggcaaaatcc gacagaggct gaattgcagg acatgattaa cgaggtagac gccgatggga
     1981 acgggactat cgactttccg gaatttctta ctatgatggc acgcaaaatg aaagataccg
     2041 attctgaaga agagatccgt gaagctttcc gtgtttttga taaagatggg aacggctaca
     2101 tcagtgctgc tgagttacgc catgtgatga caaatctggg ggaaaaactt accgacgaag
     2161 aagtagacga aatgattcgc gaggcggata ttgacgggga tggacaagta aactacgagg
     2221 aatttgtgca gatgatgacc gccaagattt ccggcggcgg aagcggcatg gtctctaagg
     2281 gggaggaact ctttaccggt gtggtgccaa tacttgtaga gctcgatggg gacgttaatg
     2341 gacacaaatt ttctgtctca ggtgagggtg agggagatgc aacttatgga aaactgacat
     2401 tgaaattcat ctgcacaaca ggcaaattgc ctgtcccctg gcctaccctg gtaaccaccc
     2461 tcacttatgg cgttcagtgc ttctcccggt atcctgatca catgaaacag catgatttct
     2521 tcaaatcagc aatgcccgaa ggttatgttc aagagcggac catctttttc aaggacgatg
     2581 gaaactataa aaccagagct gaggttaagt tcgagggaga tactttggtg aatcggatcg
     2641 aactgaaggg gatagacttc aaagaggatg ggaacatttt ggggcataaa ctcgaatata
     2701 actacaactc ccacaacgtg tatatcatgg ctgacaaaca gaagaacggg atcaaagtaa
     2761 attttaagat acgacacaat atagaggacg gtagtgtcca acttgctgat cactaccagc
     2821 agaatacacc catcggagac ggacccgttc tcttgcccga taatcactat ctctctacac
     2881 aatccgctct gtctaaagat ccaaacgaaa agcgggacca catggtactg cttgagttcg
     2941 ttacagccgc tggtatcacc ttgggcatgg atgagttgta taaataaaag cttatcgata
     3001 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3061 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3121 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3181 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3241 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3301 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3361 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3421 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3481 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3541 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3601 gacctcgacc cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     3661 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     3721 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3781 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     3841 aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc
     3901 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3961 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4021 gcggcctcag tgagcgagcg agcgcgcagc tggcgtaata gcgaagaggc ccgcaccgat
     4081 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     4141 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4201 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4261 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4321 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4381 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4441 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4501 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4561 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4621 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4681 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4741 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4801 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4861 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4921 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4981 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5041 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5101 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5161 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5221 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5281 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5341 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5401 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5461 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5521 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5581 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5641 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5701 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5761 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5821 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5881 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5941 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6001 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6061 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6121 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6181 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6241 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6301 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6361 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6421 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6481 gcgagtcgct attacgccag
//
