LOCUS       sequence-379318-        6278 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express the high-affinity histamine sensor GRAB_HA1h in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6278)
  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 6278)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_HA1h
FEATURES             Location/Qualifiers
     source          1..6278
                     /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..649
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     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     1567..1588
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1694..1715)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1955..1974)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2405..2993
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2458..2478)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2876..2887)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3025..3501
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3224..3243)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3541..3681
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3541..3670
                     /label=AAV2 ITR
     rep_origin      3756..4211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3843..3862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4053..4074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4228..4247)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4347..4369
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4407..4425)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4493..4597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4598..4666,4667..5458)
                     /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(4816..4835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5629..6217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6118..6137
                     /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 ggcaccccag atactaatag cacaatcaat ttatcactaa gcactcgtgt
      721 tactttagca ttttttatgt ccttagtagc ttttgctata atgctaggaa atgctttggt
      781 cattttagct tttgtggtgg acaaaaacct tagacatcga agtagttatt tttttcttaa
      841 cttggccatc tctgacttct ttgtgggtgt gatctccatt cctttgtaca tccctcacac
      901 gctgttcgaa tgggattttg gaaaggaaat ctgtgtattt tggctcacta ctgactatct
      961 gttatgtaca gcatctgtat ataacattgt cctcatcagc tatgatcgat acctgtcagt
     1021 ctcaaatgct gtgtcttata gaactcaaca tactggggtc ttgaagattg ttactctgat
     1081 ggtggccgtt tgggtgctgg ccttcttagt gaatgggcca atgattctag tttcagagtc
     1141 ttggaaggat gaaggtagtg aatgtgaacc tggatttttt tcggaatggt acatccttgc
     1201 catcacatca ttcttggaat tcgtgatccc agtcatctta gtcgcttatt tcaacatgaa
     1261 tatttattgg agcctgtgga agcgtgatca tctcagtagg tgcctgccag caccggccgg
     1321 gccgcgcgac accgacgcgc tggacctgga ggagggagga aatgtctata tcaaggccga
     1381 caagcagaag aacggcatca aggcgaactt cgtgatccgc cacaacatcg aggacggcgg
     1441 cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1501 gcccgacaac cactacctga gcgtgcagtc caaactttcg aaagacccca acgagaagcg
     1561 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1621 gctgtacaag ggcggtaccg gagggagcat ggtgagaaag ggcgaggagc tgttcaccgg
     1681 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1741 cggcgagggt gagggcgatg ccaccgaggg caagctgacc ctgaagttca tctgcaccac
     1801 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     1861 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1921 aggctacatc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1981 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2041 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacaccggag cagcagcacg
     2101 ctggcgcgcg gcgaagtcac tggccattct cttaggggtt tttgctgttt gctgggctcc
     2161 atattctctg ttcacaattg tcctttcatt ttattcctca gcaacaggtc ctaaatcagt
     2221 ttggtataga attgcatttt ggcttcagtg gttcaattcc tttgtcaatc ctcttttgta
     2281 tccattgtgt cacaagcgct ttcaaaaggc tttcttgaaa atattttgta taaaaaagca
     2341 acctctacca tcacaacaca gtcggtcagt atcttcttaa gaattcgata tcaagcttat
     2401 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2461 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2521 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2581 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2641 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2701 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2761 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     2821 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2881 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2941 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3001 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     3061 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3121 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3181 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3241 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3301 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3361 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3421 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3481 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     3541 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3601 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3661 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3721 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3781 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3841 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3901 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3961 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4021 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4081 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4141 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4201 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4261 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4321 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4381 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4441 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4501 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4561 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4621 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4681 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4741 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4801 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4861 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4921 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4981 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5041 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5101 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5161 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5221 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5281 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5341 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5401 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5461 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5521 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5581 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5641 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5701 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5761 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5821 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5881 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5941 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6001 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6061 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6241 acggttcctg gccttttgct ggccttttgc tcacatgt
//
