LOCUS       sequence-412464-        6551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent dopamine (DA) sensor 
            GRAB_gDA3m in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6551)
  AUTHORS   Zhuo Y, Luo B, Yi X, Dong H, Miao X, Wan J, Williams JT, Campbell 
            MG, Cai R, Qian T, Li F, Weber SJ, Wang L, Li B, Wei Y, Li G, Wang 
            H, Zheng Y, Zhao Y, Wolf ME, Zhu Y, Watabe-Uchida M, Li Y
  TITLE     Improved green and red GRAB sensors for monitoring dopaminergic 
            activity in vivo.
  JOURNAL   Nat Methods. 2023 Nov 30. doi: 10.1038/s41592-023-02100-w.
  PUBMED    38036855
REFERENCE   2  (bases 1 to 6551)
  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: "Nat 
            Methods. 2023 Nov 30. doi: 10.1038/s41592-023-02100-w."
COMMENT     Sequence Label: pAAV-hSyn-GRAB-gDA3m
FEATURES             Location/Qualifiers
     source          1..6551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..142
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      593..602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     599..658
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    662..686
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1585..1606
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1712..1733)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2690..3278
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2743..2763)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3161..3172)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3310..3786
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3509..3528)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3826..3966
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3826..3955
                     /label=AAV2 ITR
     rep_origin      4041..4496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4128..4147)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4338..4359
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4513..4532)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4632..4654
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4692..4710)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4778..4882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4883..4951,4952..5743)
                     /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(5101..5120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5914..6502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6403..6422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctgcggccgc acgcgtatga ggcggggtgg gggtgcctac
      181 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      241 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      301 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      361 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      421 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      481 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      541 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgagaaaccg gtgccaccat
      601 ggagacagac acactcctgc tatgggtact gctgctctgg gttccaggtt ccactggtga
      661 cacaagtttg tacaaaaaag ttggcaccac cggtaggact ctgaacacct ctgccatgga
      721 cgggactggg ctggtggtgg agagggactt ctctgttcgt atcctcactg cctgtttcct
      781 gtcgctgctc atcctgtcca cgctcctggg gaacacgctg gtctgtgctg ccgttatcag
      841 gttccgacac ctgcggtcca aggtgaccaa cttctttgtc atctccttgg ctgtgtcaga
      901 tctcttggtg gccgtcctgg tcatgccctg gaaggcagtg gctgagattg ctggcttctg
      961 gccctttggg tccttctgta acatctgggt ggcctttgac atcatgtgct ccactgcatc
     1021 catcctcaac ctctgtgtga tcagcgtgga caggtattgg gctatctcca gccctgcccg
     1081 gtatgagaga aagatgaccc ccaaggcagc cttcatcctg atcagtgtgg catggacctt
     1141 gtctgtactc atctccttca tcccagtgca gctcagctgg cacaaggcaa aacccacaag
     1201 cccctctgat ggaaatgcca cttccctggc tgagaccata gacaactgtg actccagcct
     1261 cagcaggaca tatgccatct catcctctgt aataagcttt tacatccctg tggccatcat
     1321 gattgtcttg tacaccagga tctacaggat tgctcagaaa cagctgaatt cactcattta
     1381 tgtctatatc aaggccgaca agcagaagaa cggcatcaag gcgaacttcg ggatccgcca
     1441 caacatcgag gacggtggcg tgcagctcgc ctaccactac cagcagaaca cccccatcgg
     1501 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca aactttcgaa
     1561 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1621 cactctcggc atggacgagc tgtacaaggg cggtaccgga gggctgatgg tgagcaaggg
     1681 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1741 ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc acctacggca agctgaccct
     1801 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1861 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1921 caagtccgcc atgcccgaag gctacatcca ggagcgcacc atcttcttca agaatgacgg
     1981 cttttacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2041 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2101 caatcatgac caactgaaaa gagaaactaa agtcctgaag actctgtcgg tgatcatggg
     2161 tgtgtttgtg tgctgttggc tacctttctt catcttgaac tgcattttgc ccttctgtgg
     2221 gtctggggag acgcagccct tctgcattga ttccaacacc tttgacgtgt ttgtgtggtt
     2281 tgggtgggct aattcatcct tgaaccccat catttatgcc tttaatgctg attttcggaa
     2341 ggcattttca accctcttag gatgctacag actttgccct gcgacgaata atgccataga
     2401 gacggtgagt atcaataaca atggggccgc gatgttttcc agccatcatg agccacgagg
     2461 ctccatctcc aaggagtgca atctggttta cctgatccca catgctgtgg gctcctctga
     2521 ggacctgaaa aaggaggagg cagctggcat cgccagaccc ttggagaagc tgtccccagc
     2581 cctatcggtc atattggact atgacactga cgtctctctg gagaagatcc aacccatcac
     2641 acaaaacggt cagcacccaa cctgagaatt cgatatcaag cttatcgata atcaacctct
     2701 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2761 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2821 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2881 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2941 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3001 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3061 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     3121 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3181 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3241 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3301 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3361 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3421 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3481 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     3541 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     3601 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     3661 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     3721 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     3781 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3841 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3901 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3961 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4021 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4081 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4141 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4201 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4261 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4321 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4381 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4441 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4501 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4561 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4621 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4681 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4741 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4801 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4861 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4921 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4981 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5041 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5101 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5161 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5221 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5281 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5341 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5401 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5461 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5521 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5581 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5641 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5701 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5761 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5821 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5881 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5941 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6001 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6061 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6121 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6181 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6241 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6301 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6361 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6421 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6481 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6541 ttgctggcct t
//
