LOCUS       sequence-217601-        6338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express the genetically-encoded fluorescent dopamine(DA) sensor 
            GRAB_DA1h in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6338)
  AUTHORS   Sun F, Zeng J, Jing M, Zhou J, Feng J, Owen SF, Luo Y, Li F, Wang H,
            Yamaguchi T, Yong Z, Gao Y, Peng W, Wang L, Zhang S, Du J, Lin D, Xu
            M, Kreitzer AC, Cui G, Li Y
  TITLE     A Genetically Encoded Fluorescent Sensor Enables Rapid and Specific 
            Detection of Dopamine in Flies, Fish, and Mice.
  JOURNAL   Cell. 2018 Jul 12;174(2):481-496.e19. doi: 
            10.1016/j.cell.2018.06.042.
  PUBMED    30007419
REFERENCE   2  (bases 1 to 6338)
  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: 
            "10.1016/j.cell.2018.06"; journalName: "Cell"; date: "2018-07-12- 
            12"; volume: "174"; issue: "2"; pages: "481-496.e19"
COMMENT     Sequence Label: pAAV-hSyn-GRAB_DA1h
FEATURES             Location/Qualifiers
     source          1..6338
                     /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     1648..1669
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1775..1796)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2036..2055)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2518..2538)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2936..2947)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3085..3561
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3284..3303)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3601..3741
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3601..3730
                     /label=AAV2 ITR
     rep_origin      3816..4271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3903..3922)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4113..4134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4288..4307)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4407..4429
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4467..4485)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4553..4657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4658..4726,4727..5518)
                     /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(4876..4895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5689..6277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6178..6197
                     /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 gtgatccact gaatctgtcc tggtatgatg atgatctgga
      721 gaggcagaac tggagccggc ccttcaacgg gtcagacggg aaggcggaca gaccccacta
      781 caactactat gccacactgc tcaccctgct catcgctgtc atcgtcttcg gcaacgtgct
      841 ggtgtgcatg gctgtgtccc gcgagaaggc gctgcagacc accaccaact acctgatcgt
      901 cagcctcgca gtggccgacc tcctcgtcgc cacactggtc atgccctggg ttgtctacct
      961 ggaggtggta ggtgagtgga aattcagcag gattcactgt gacatcttcg tcactctgga
     1021 cgtcatgatg tgcacggcga gcatcctgaa cttgtgtgcc atcagcatcg acaggtacac
     1081 agctgtggcc atgcccatgc tgtacaatac gcgctacagc tccaagcgcc gggtcaccgt
     1141 catgatctcc atcgtctggg tcctgtcctt caccatctcc tgcccactcc tcttcggact
     1201 caataacgca gaccagaacg agtgcatcat tgccaacccg gccttcgtgg tctactcctc
     1261 catcgtctcc ttctacgtgc ccttcattgt catgctgctg gtctacatca agatctacat
     1321 tgtcctccgc agacgccgca agcgagtcaa caccaaacgc agcagccgag ctttcagggc
     1381 ccacctgagg gctccactaa agggcaactg tactcacccc gaggacatga aactcggagg
     1441 aaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact tccacatccg
     1501 ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat
     1561 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt ccaaactttc
     1621 gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1681 gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca tggtgagcaa
     1741 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1801 cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac
     1861 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1921 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1981 cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga
     2041 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2101 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2161 caacactgga gcagcagcaa tgagccgtag gaagctctcc cagcagaagg agaagaaagc
     2221 cactcagatg ctcgccattg ttctcggcgt gttcatcatc tgctggctgc ccttcttcat
     2281 cacacacatc ctgaacatac actgtgactg caacatcccg cctgtcctgt acagcgcctt
     2341 cacgtggctg ggctatgtca acagcgccgt gaaccccatc atctacacca ccttcaacat
     2401 tgagttccgc aaggccttcc tgaagatcct ccactgctga gaattcgata tcaagcttat
     2461 cgatcatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2521 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2581 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2641 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2701 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2761 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2821 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     2881 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2941 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3001 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3061 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     3121 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3181 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3241 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3301 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3361 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3421 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3481 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3541 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     3601 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3661 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3721 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3781 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3841 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3901 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3961 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4021 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4081 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4141 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4201 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4261 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4321 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4381 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4441 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4501 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4621 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4681 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4741 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4801 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4861 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4981 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5041 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5221 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5281 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5341 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5521 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5581 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5641 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5701 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5761 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5821 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5881 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5941 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6001 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6061 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6121 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6181 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6241 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6301 acggttcctg gccttttgct ggccttttgc tcacatgt
//
