LOCUS       sequence-379319-        6539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the medium-affinity histamine sensor GRAB_HA1m in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6539)
  AUTHORS   Dong H, Li M, Yan Y, Qian T, Lin Y, Ma X, Vischer HF, Liu C, Li G, 
            Wang H, Leurs R, Li Y
  TITLE     Genetically encoded sensors for measuring histamine release both in 
            vitro and in vivo.
  JOURNAL   Neuron. 2023 Mar 7:S0896-6273(23)00128-9. doi: 
            10.1016/j.neuron.2023.02.024.
  PUBMED    36924772
REFERENCE   2  (bases 1 to 6539)
  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"; journalName: "Neuron. 
            2023 Mar 7:S0896-6273(23)00128-9. doi: 
            10.1016/j.neuron.2023.02.024."
COMMENT     Sequence Label: pAAV-hsyn-GRAB_HA1m
FEATURES             Location/Qualifiers
     source          1..6539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     587..646
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1726..1747
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1853..1874)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2114..2133)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2666..3254
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2719..2739)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3137..3148)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3286..3762
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3485..3504)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3802..3942
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3802..3931
                     /label=AAV2 ITR
     rep_origin      4017..4472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4104..4123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4314..4335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4489..4508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4608..4630
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4668..4686)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4754..4858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4859..4927,4928..5719)
                     /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(5077..5096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5890..6478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6379..6398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
      181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac
      601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
      661 caaaaaagtt ggcaccaccg gtatggagcc cgccggcgcc ggcggcgccg ccattctgct
      721 ggacgacgac gacgactact acctcacaac ccccggctac cccggctccg gctctaacgc
      781 ctccctgtct cctccagccg acctgttccc cgccgcctct aaccactcga ccgctgtcgt
      841 ggtgtctggc gacctggacc tgtacgtgat cattccactg ggcatcttcc tcgccgtgct
      901 gtctctcgtg acattcgccg gcaacgtgat ggtgctgcac gccctcagaa cagagaagaa
      961 gctccaaaca gtgtctaaca ttttcatcat gtctctcgcc attgccgact gctctgtggg
     1021 cctgttcgcc atgccactga gcgccgtgta cgccctgctg ggctactggc ctttcggcct
     1081 cgtggtgtgc aacatgtggt tgagtatgga ctacgtggcc agcacagcct ctattttcaa
     1141 cctgttcgcc ctgtccctcg accggttctg gtctattatt tctccactca gatacctcaa
     1201 gaagcggaca cgcaagcggg ccattattat gtgctctagc gtgtggctcg cctctctcct
     1261 gtggattatc ccggttgtag gatggcacca gatgttcaac gacggcaggc gggacgtgcc
     1321 agtggaggag tgcgacaccg agttcgccga cgagcaggtg ttcaaggtgc tgacagccat
     1381 cttcaacttc tacgtgccta tggccgccat gattgccctg tacggcagga ttttccacga
     1441 gattaagctc agagccgcgt ctcagccagc accggccggg ccgcgcgaca ccgacgcgct
     1501 ggacctggag gagggaggaa acgtctatat caaggccgac aagcagaaga acggcatcaa
     1561 ggcgaacttc gtgatccgcc acaacatcga ggacggcggc gtgcagctcg cctaccacta
     1621 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1681 cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1741 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg
     1801 agggagcatg gtgagaaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1861 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc
     1921 caccgagggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1981 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2041 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac
     2101 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2161 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2221 ggggcacaag ctggagtaca acaccggagc agcagcacgc tggcgcgcga agaaggccgc
     2281 cttccagctg ggcgtgatcc tgggcgcctt ctgcctgtgc ttcatgcctt acttcattag
     2341 cttcatcatt accgccttct gcaaggactg cgtgtctcct gccgtgcact ctattttcac
     2401 atggctgggc tacgtgaact ctacaacaaa ccctttcctg tacacatgga caaacgccaa
     2461 cttccggtcc gccttccgga agatcctccg gatgaaggag acaccaaaga agccagtgta
     2521 catgcagatg cctcacaaga cccctagacc aaagaagcag ccattagaag ctagggtgag
     2581 cggcacagcc tctggcggca ccacagcccc agcccgcaag tctgacgaca ccagactcta
     2641 agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     2701 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2761 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2821 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2881 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2941 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3001 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3061 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     3121 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3181 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3241 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt
     3301 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     3361 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     3421 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     3481 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     3541 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     3601 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     3661 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     3721 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     3781 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     3841 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3901 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     3961 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4021 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4081 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4141 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4201 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4261 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4321 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     4381 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4441 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     4501 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     4561 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4621 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     4681 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4741 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4801 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4861 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4921 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4981 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5041 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5101 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5161 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5221 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5281 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5341 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5401 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5461 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5521 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5581 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5641 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5701 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5761 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5821 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5881 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5941 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6001 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6061 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6121 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6181 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6241 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6301 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6361 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6421 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6481 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
