LOCUS       sequence-412473-        6320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent dopamine (DA) control 
            sensor GRAB_rDA2mut 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-rDA2mut
FEATURES             Location/Qualifiers
     source          1..6320
                     /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     2145..2164
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2461..3049
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2514..2534)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2932..2943)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3081..3557
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3280..3299)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3597..3737
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3597..3726
                     /label=AAV2 ITR
     rep_origin      3812..4267
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3899..3918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4109..4130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4284..4303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4403..4425
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4463..4481)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4549..4653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4654..4722,4723..5514)
                     /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(4872..4891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5685..6273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6174..6193
                     /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 accggtatca accgactgaa ctacagcgtc
      721 ctcgtgaacc tgacggccta ctacgacggc ggcgcgggtc tgaatctcag cagcgtcaat
      781 tgcacgtcgt cgatagtcgc cgggggtgcc gttagaccgg gcgagtgcgc tggaaccgcc
      841 ggcgtcgacg agaagtctaa cgcgaacagc tggtgggccc tgatacttgt gatcgtgccg
      901 tgtttgacgc tcttcggtaa tgtgctcgtc atcctagcgg tggtgaaaga acgaacgctg
      961 cagactgtca ctaactactt catcgtcagt ttggcggtcg ccgatcttct cgtggcggtg
     1021 ctcgttatgc cgttcgcggt ctacgtgctg gtgaacggat cctggagttt acccgggttt
     1081 gtttgtgatt tttatatcgc aatggatgtg acttgcgcga ccagctctat attcaatctc
     1141 gtggctattt caatagacag atacatagcg gtgacccagc cgataaagta cgcgaagcac
     1201 aagaacaata gaagagtatg gttgacgata ctgttggtct gggcgatatc ggccgcgatc
     1261 ggcagcccga ttgtcctagg cttgaataac actccggacc ggatcccgga cgcatgtctg
     1321 ttctacaata cggatttcat catctacaat agcctgtcca gcttctacat accctgcatc
     1381 atcatggtat tcctttacta caatatattc aaggctctac gaaacagagc gaaacttttt
     1441 cgtgctagct gcgtgtgttc cgaggttatg taccccgagg acggcgccct gaagagcgag
     1501 atctggaagg ggctgaggct gaaggacggc ggccactacg ccgccgaggt caagaccacc
     1561 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca tcgtcgacat caagttggac
     1621 atcgtgtccc acaacgagga ctacaccatc gtggaacagt gcgaacgcgc cgaggcccgc
     1681 cactccaccg gcggcatgga cgagctgtac aagggaggta caggcgggag tctggtgagc
     1741 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     1801 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1861 gaggcctttc agaccgctaa gctgaaggtg accaagggtg gccccctgcc cttcacttgg
     1921 gacatcctgt cccctcagtt catgtacggc tccaaggcct acattaagca cccagccgac
     1981 atccccgact acttcaagct gtccttcccc gagggcttca ggtgggagcg cgtgatgaac
     2041 ttcgaggacg gcggcattat tcacgttaac caggactcct ccctgcagga cggctacttc
     2101 atctacaagg tgaagctgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     2161 aagaccatgg gctgggagct gacggtcctc gagcgcaagg ccacgaagcc tctggcgatc
     2221 gtgttaggtg tctttctaat atgctgggtg cccttcttca cttgtaacat catggatgca
     2281 atctgcacga agctgacgaa ggcttgtcag cctggcgtta cagccttcat cgtcacttcc
     2341 tggttgggct acatgaacag cttcgtaaat cccgtaatat acactgtgtt caaccccgaa
     2401 ttccgtaagg ccttccgcaa gctgatcagc atataagaat tcgatatcaa gcttatcgat
     2461 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2521 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2581 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2641 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2701 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2761 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2821 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     2881 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     2941 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3001 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     3061 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     3121 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     3181 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     3241 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     3301 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     3361 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     3421 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     3481 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     3541 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     3601 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3661 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3721 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3781 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4021 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4141 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     4201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4261 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4321 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4381 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4441 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4501 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4621 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4681 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4741 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4801 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4861 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4921 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4981 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5041 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5101 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5161 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5221 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5281 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5341 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5401 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5461 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5521 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5581 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5701 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5761 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5821 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5881 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5941 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6001 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6061 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6301 ttcctggcct tttgctggcc
//
