LOCUS       sequence-412470-        6809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent dopamine (DA) sensor 
            GRAB_rDA3m in neurons in a cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6809)
  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 6809)
  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-DIO-GRAB-rDA3m
FEATURES             Location/Qualifiers
     source          1..6809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(602..635)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(686..719)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2791..2824
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2875..2908
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2933..3521
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2986..3006)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3404..3415)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3553..4029
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3752..3771)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4069..4209
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4069..4198
                     /label=AAV2 ITR
     rep_origin      4284..4739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4371..4390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4581..4602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4756..4775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4875..4897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4935..4953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5021..5125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5126..5194,5195..5986)
                     /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(5344..5363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6157..6745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6646..6665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6807..6809,1..127)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
       61 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      121 ggttcctgcg gccgcacgcg tatgaggcgg ggtgggggtg cctacctgac gaccgacccc
      181 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg
      241 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca
      301 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg
      361 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc
      421 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac
      481 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga
      541 gtcgtgtcgt gcctgagagc gcagtcgaga aaccggtgcc acctctagag tcgactccgg
      601 aataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct
      661 gctattagca atatgaaacc tcttaataac ttcgtatagc atacattata cgaagttatg
      721 gcgcatatgg gtcaggttgg gtgctgaccg ttttgtgtga tgggttggat cttctccaga
      781 gagacgtcag tgtcatagtc caatatgacc gatagggctg gggacagctt ctccaagggt
      841 ctggcgatgc cagctgcctc ctcctttttc aggtcctcag aggagcccac agcatgtggg
      901 atcaggtaaa ccagattgca ctccttggag atggagcctc gtggctcatg atggctggaa
      961 aacatcgcgg ccccattgtt attgatactc accgtctcta tggcattatt cgtcgcaggg
     1021 caaagtctgt agcatcctaa gagggttgaa aatgccttcc gaaaatcagc attaaaggca
     1081 taaatgatgg ggttcaagga tgaattagcc cacccaaacc acacaaacac gtcaaaggtg
     1141 ttggaatcaa tgcagaaggg ctgcgtctcc ccagacccac agaagggcaa aatgcagttc
     1201 aagatgaaga aaggtagcca acagcacaca aacacaccca tgatcaccga cagagtcttc
     1261 aggactttag tttctctttt aaggtcgtcc ttccgatgtt cccaccccat agttttcttc
     1321 tgcattacgg ggccgtcggg ggggaagttg gtgccgcgca gcttcacctt gtagatgaat
     1381 acgccgtcct gcagggagga gtcctggttc acgtgaataa tgccgccgtc ctcgaagttc
     1441 atcacgcgct cccacctgaa gccctcgggg aaggacgact tgaagtagtc ggggatgtcg
     1501 gctgggtgct taatgtaggc cttggagccg tacatgaact gaggggacag gatgtcccag
     1561 gcgaagggca gggggccacc cttggtcacc ttcagcttag cggtctgaaa ggcctcgtag
     1621 gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg
     1681 tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctc
     1741 accagactcc cgcctgtacc tcccttgtac agctcgtcca tgccgccggt ggagtggcgg
     1801 ccctcggcgc gttcgcactg ttccacgatg gtgtagtcct cgttgtggga cacgatgtcc
     1861 aacttgatgt ccacgatgta ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg
     1921 gtcttgacct cggcggcgta gtggccgccg tccttcagcc tcagcccctt cttgatctcg
     1981 ctcttcaggg cgccgtcctc ggggtacatc cgctcggaaa ccatgaagga ggagagcttt
     2041 ttctgagcaa tcctgtagat cctggtgtag cagacaatca tgatggccac agggatgtaa
     2101 aagcttatta cagaggatga gatggcatat gtcctgctga ggctggagtc acagttgtct
     2161 atggtctcag ccagggaagt ggcatttcca tcagaggggc ttgtgggttt tgccttgtgc
     2221 cagctgagct gcactgggat gaaggagatg agtacagaca aggtccatgc cacactgatc
     2281 aggatgaagg ctgccttggg ggtcatcttt ctctcatacc gctcagggct ggagatagcc
     2341 caatacctgt ccacgctgat cacacagagg ttgaggatgg atgcagtgga gcacatgatg
     2401 tcaaaggcca cccagatgtt acagaaggac ccaaagggcc agaagccagc aatctcagcc
     2461 actgccttcc agggcatgac caggacggcc accaagagat ctgacacagc caaggagatg
     2521 acaaagaagt tggtcacctt ggaccgcagg tgtcggaacc tgataacggc agcacagacc
     2581 agcgtgttcc ccaggagcgt ggacaggatg agcagcgaca ggaaacaggc agtgaggata
     2641 cgaacagaga agtccctctc caccaccagc ccagtcccgt ccatggcaga ggtgttcaga
     2701 gtcctcatgg cgtcatcatc gtccttgtag tcggcgaaca ccaagcagaa gatgtagctc
     2761 agggcgatga tcgtcttcat tggcgctagc ataacttcgt ataaagtatc ctatacgaag
     2821 ttatttgcct taacccagaa attatcactg ttattcttta gaatggtgca aagaataact
     2881 tcgtataatg tatgctatac gaagttatga attcgatatc aagcttatcg ataatcaacc
     2941 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3001 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3061 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3121 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3181 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3241 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3301 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3361 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3421 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3481 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     3541 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     3601 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     3661 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     3721 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     3781 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     3841 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     3901 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     3961 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     4021 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     4081 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4141 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4201 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4261 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4321 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4381 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4441 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4501 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4561 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4621 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4681 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4741 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4801 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4861 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4921 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4981 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5041 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5101 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5161 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5221 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5281 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5341 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5401 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5461 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5521 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5581 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5641 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5701 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5761 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5821 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5881 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5941 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6001 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6061 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6121 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6181 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6241 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6301 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6361 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6421 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6481 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6541 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6601 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6661 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6721 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6781 cttttgctgg ccttttgctc acatgtcct
//
