LOCUS       sequence-412472-        6551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent dopamine (DA) control 
            sensor GRAB_gDA3mut 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-gDA3mut
FEATURES             Location/Qualifiers
     source          1..6551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   15..144
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     601..660
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    664..688
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1587..1608
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1714..1735)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2692..3280
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2745..2765)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3163..3174)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3312..3788
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3511..3530)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3828..3968
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3828..3957
                     /label=AAV2 ITR
     rep_origin      4043..4498
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4130..4149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4340..4361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4515..4534)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4634..4656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4694..4712)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4780..4884
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4885..4953,4954..5745)
                     /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(5103..5122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5916..6504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6405..6424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
       61 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
      121 ccaactccat cactaggggt tcctgcggcc gcacgcgtat gaggcggggt gggggtgcct
      181 acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc
      241 atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag
      301 cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca
      361 gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc
      421 ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag
      481 gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg
      541 cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcgagaaac cggtgccacc
      601 atggagacag acacactcct gctatgggta ctgctgctct gggttccagg ttccactggt
      661 gacacaagtt tgtacaaaaa agttggcacc accggtagga ctctgaacac ctctgccatg
      721 gacgggactg ggctggtggt ggagagggac ttctctgttc gtatcctcac tgcctgtttc
      781 ctgtcgctgc tcatcctgtc cacgctcctg gggaacacgc tggtctgtgc tgccgttatc
      841 aggttccgac acctgcggtc caaggtgacc aacttctttg tcatctcctt ggctgtgtca
      901 gatctcttgg tggccgtcct ggtcatgccc tggaaggcag tggctgagat tgctggcttc
      961 tggccctttg ggtccttctg taacatctgg gtggcctttg acatcatgtg ctccactgca
     1021 tccatcctca acctctgtgt gatcagcgtg gacaggtatt gggctatctc cagccctgcc
     1081 cggtatgaga gaaagatgac ccccaaggca gccttcatcc tgatcagtgt ggcatggacc
     1141 ttgtctgtac tcatctcctt catcccagtg cagctcagct ggcacaaggc aaaacccaca
     1201 agcccctctg atggaaatgc cacttccctg gctgagacca tagacaactg tgactccagc
     1261 ctcagcagga catatgccat ctcagcctct gtaataccct tttacatccc tgtggccatc
     1321 atgattgtct tgtacaccag gatctacagg attgctcaga aacagctgaa ttcactcatt
     1381 tatgtctata tcaaggccga caagcagaag aacggcatca aggcgaactt cgggatccgc
     1441 cacaacatcg aggacggtgg cgtgcagctc gcctaccact accagcagaa cacccccatc
     1501 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc caaactttcg
     1561 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1621 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggctgat ggtgagcaag
     1681 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1741 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     1801 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1861 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1921 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caagaatgac
     1981 ggcttttaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2041 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2101 aacaatcatg accaactgaa aagagaaact aaagtcctga agactctgtc ggtgatcatg
     2161 ggtgtgtttg tgtgctgttg gctacctttc ttcatcttga actgcatttt gcccttctgt
     2221 gggtctgggg agacgcagcc cttctgcatt gattccaaca cctttgacgt gtttgtgtgg
     2281 tttgggtggg ctaattcatc cttgaacccc atcatttatg cctttaatgc tgattttcgg
     2341 aaggcatttt caaccctctt aggatgctac agactttgcc ctgcgacgaa taatgccata
     2401 gagacggtga gtatcaataa caatggggcc gcgatgtttt ccagccatca tgagccacga
     2461 ggctccatct ccaaggagtg caatctggtt tacctgatcc cacatgctgt gggctcctct
     2521 gaggacctga aaaaggagga ggcagctggc atcgccagac ccttggagaa gctgtcccca
     2581 gccctatcgg tcatattgga ctatgacact gacgtctctc tggagaagat ccaacccatc
     2641 acacaaaacg gtcagcaccc aacctgagaa ttcgatatca agcttatcga taatcaacct
     2701 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2761 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2821 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2881 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2941 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3001 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3061 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     3121 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3181 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3241 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     3301 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3361 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3421 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3481 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3541 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3601 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3661 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     3721 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3781 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     3841 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3901 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3961 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4021 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4081 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4141 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4201 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4261 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4321 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4381 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4441 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4501 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4561 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4621 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4681 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4741 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4801 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4861 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4921 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4981 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5041 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5101 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5161 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5221 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5281 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5341 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5401 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5461 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5521 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5581 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5641 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5701 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5761 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5821 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5881 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5941 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6001 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6061 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6121 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6181 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6241 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6301 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6361 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6421 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6481 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6541 ttttgctggc c
//
