LOCUS       sequence-412466-        6320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent dopamine (DA) sensor 
            GRAB_rDA2m in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6320)
  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 6320)
  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-rDA2m
FEATURES             Location/Qualifiers
     source          1..6320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(82..186)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     254..272
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(310..332)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     432..451
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(468..923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(605..626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     817..836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   998..1138
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1009..1138
                     /label=AAV2 ITR
     polyA_signal    1178..1654
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1436..1455
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             1792..1803
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2201..2221
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(2571..2590)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(4047..4071)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(4075..4134)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      4131..4140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4591..4720
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4782..5370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4862..4881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5541..6320,1..12,13..81))
                     /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     6164..6183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
       61 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      121 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      181 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
      241 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
      301 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
      361 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
      421 gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt
      481 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
      541 gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt
      601 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
      661 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
      721 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
      781 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
      841 cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
      901 taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg tgaaataccg
      961 cacagatgcg taaggagaaa ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc
     1021 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     1081 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     1141 ggccgctcgg tccgcacgtg gttacctaca aaatcagaag gacagggaag ggagcagtgg
     1201 ttcacgcctg taatcccagc aatttgggag gccaaggtgg gtagatcacc tgagattagg
     1261 agttggagac cagcctggcc aatatggtga aaccccgtct ctaccaaaaa aacaaaaatt
     1321 agctgagcct ggtcatgcat gcctggaatc ccaacaactc gggaggctga ggcaggagaa
     1381 tcgcttgaac ccaggaggcg gagattgcag tgagccaaga ttgtgccact gcactccagc
     1441 ttggttccca atagaccccg caggccctac aggttgtctt cccaacttgc cccttgctcc
     1501 ataccacccc cctccacccc ataatattat agaaggacac ctagtcagac aaaatgatgc
     1561 aacttaattt tattaggaca aggctggtgg gcactggagt ggcaacttcc agggccagga
     1621 gaggcactgg ggaggggtca cagggatgcc acccgtagat ctctcgagca gcgctcggta
     1681 tcgatgcggg gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg
     1741 cggaagaggc cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc
     1801 cgaagggacg tagcagaagg acgtcccgcg cagaatccag gtggcaacat aggcgagcag
     1861 ccaaggaaag gacgatgatt tccccgacaa caccacggaa ttgtcagtgc ccaacagccg
     1921 agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag
     1981 ggggaaagcg aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg
     2041 ggttgcgtca gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc
     2101 ctcataaaga gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga
     2161 agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac
     2221 atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt gatgatcgat
     2281 aagcttgata tcgaattctt atatgctgat cagcttgcgg aaggccttac ggaattcggg
     2341 gttgaacaca gtgtatatta cgggatttac gaagctgttc atgtagccca accaggaagt
     2401 gacgatgaag gctgtaacgc caggctgaca agccttcgtc agcttcgtgc agattgcatc
     2461 catgatgtta caagtgaaga agggcaccca gcatattaga aagacaccta acacgatcgc
     2521 caggggcttc gtggccttgc gctcgaggac cgtcagctcc cagcccatgg tcttcttctg
     2581 cattacgggg ccgtcggggg ggaagttggt gccgcgcagc ttcaccttgt agatgaagta
     2641 gccgtcctgc agggaggagt cctggttaac gtgaataatg ccgccgtcct cgaagttcat
     2701 cacgcgctcc cacctgaagc cctcggggaa ggacagcttg aagtagtcgg ggatgtcggc
     2761 tgggtgctta atgtaggcct tggagccgta catgaactga ggggacagga tgtcccaagt
     2821 gaagggcagg gggccaccct tggtcacctt cagcttagcg gtctgaaagg cctcgtaggg
     2881 gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg ttcacggagc cctccatgtg
     2941 caccttgaag cgcatgaact ccttgatgat ggccatgtta tcctcctcgc ccttgctcac
     3001 cagactcccg cctgtacctc ccttgtacag ctcgtccatg ccgccggtgg agtggcgggc
     3061 ctcggcgcgt tcgcactgtt ccacgatggt gtagtcctcg ttgtgggaca cgatgtccaa
     3121 cttgatgtcg acgatgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt
     3181 cttgacctcg gcggcgtagt ggccgccgtc cttcagcctc agccccttcc agatctcgct
     3241 cttcagggcg ccgtcctcgg ggtacataac ctcggaacac acgcagctag cacgaaaaag
     3301 tttcgctctg tttcgtagag ccttgaatat attgtagtaa aggaatacca tgatgatgca
     3361 gggtatgtag aagctggaca ggctcgagta gatgatgaaa tccgtattgt agaacagaca
     3421 tgcgtccggg atccggtccg gagtgttatt caagcctagg acaatcgggc tgccgatcgc
     3481 ggccgatatc gcccagacca acagtatcgt caaccatact cttctattgt tcttgtgctt
     3541 cgcgtacttt atcggctggg tcaccgctat gtatctgtct attgaaatag ccacgagatt
     3601 gaatatagag ctggtgctgc aagtcacatc cattgcgata taaaaatcac aaacaaaccc
     3661 gggtaaactc caggatccgt tcaccagcac gtagaccgcg aacggcataa cgagcaccgc
     3721 cacgagaaga tcggcgaccg ccaaactgac gatgaagtag ttagtgacag tctgcagcgt
     3781 tcgttctttc accaccgcta ggatgacgag cacattaccg aagagcgtca aacacggcac
     3841 gatcacaagt atcagggccc accagctgtt cgcgttagac ttctcgtcga cgccggcggt
     3901 tccagcgcac tcgcccggtc taacggcacc cccggcgact atcgacgacg tgcaattgac
     3961 gctgctgaga ttcagacccg cgccgccgtc gtagtaggcc gtcaggttca cgaggacgct
     4021 gtagttcagt cggttgatac cggtggtgcc aacttttttg tacaaacttg tgtcaccagt
     4081 ggaacctgga acccagagca gcagtaccca tagcaggagt gtgtctgtct ccatggtggc
     4141 accggtttct cgactgcgct ctcaggcacg acacgactcc tccgctgccc accgcagact
     4201 gaggcagcgc tgagtcgccg gcgccgcagc gcagatggtc gcgcccgtgc ccccctatct
     4261 cgcgcctcgc gtggtgcggt ccggctgggc cggcggcggc gcggacgcga ccaaggtggc
     4321 cgggaagggg agtttgcggg ggaccggcga gtgacgtcag cgcgccttca gtgctgaggc
     4381 ggcggtggcg cgcgccgcca ggcgggggcg aaggcactgt ccgcggtgct gaagctggca
     4441 gtgcgcacgc gcctcgccgc atcctgtttc ccctccccct ctctgatagg ggatgcgcaa
     4501 tttggggaat gggggttggg tgcttgtcca gtgggtcggg gtcggtcgtc aggtaggcac
     4561 ccccaccccg cctcatacgc gtgcggccgc aggaacccct agtgatggag ttggccactc
     4621 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4681 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag
     4741 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4801 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4861 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4921 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4981 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5041 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5101 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5161 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5221 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5281 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5341 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5401 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5461 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5521 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5581 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5641 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5701 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5761 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5821 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5881 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5941 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6001 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6061 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6121 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6181 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6241 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6301 gcatctttta ctttcaccag
//
