LOCUS       sequence-412469-        6827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent dopamine (DA) sensor 
            GRAB_gDA3m in neurons in a cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6827)
  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 6827)
  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-gDA3m
FEATURES             Location/Qualifiers
     source          1..6827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..134
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(609..642)
                     /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(693..726)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1669..1690
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1796..1817)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2716..2740)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(2744..2803)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2800..2809
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2816..2849
                     /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    2900..2933
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2958..3546
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3011..3031)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3429..3440)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3578..4054
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3777..3796)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4094..4234
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4094..4223
                     /label=AAV2 ITR
     rep_origin      4309..4764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4396..4415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4606..4627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4781..4800)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4900..4922
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4960..4978)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5046..5150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5151..5219,5220..6011)
                     /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(5369..5388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6182..6770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6671..6690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
       61 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      121 cactaggggt tcctgcggcc gcacgcgtat gaggcggggt gggggtgcct acctgacgac
      181 cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc
      241 agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc
      301 gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg
      361 cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt
      421 ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg
      481 gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag
      541 cggaggagtc gtgtcgtgcc tgagagcgca gtcgagaaac cggtgccacc tctagagtcg
      601 actccggaat aacttcgtat aggatacttt atacgaagtt atgcagaatg gtagctggat
      661 tgtagctgct attagcaata tgaaacctct taataacttc gtatagcata cattatacga
      721 agttatggcg catatgtcag gttgggtgct gaccgttttg tgtgatgggt tggatcttct
      781 ccagagagac gtcagtgtca tagtccaata tgaccgatag ggctggggac agcttctcca
      841 agggtctggc gatgccagct gcctcctcct ttttcaggtc ctcagaggag cccacagcat
      901 gtgggatcag gtaaaccaga ttgcactcct tggagatgga gcctcgtggc tcatgatggc
      961 tggaaaacat cgcggcccca ttgttattga tactcaccgt ctctatggca ttattcgtcg
     1021 cagggcaaag tctgtagcat cctaagaggg ttgaaaatgc cttccgaaaa tcagcattaa
     1081 aggcataaat gatggggttc aaggatgaat tagcccaccc aaaccacaca aacacgtcaa
     1141 aggtgttgga atcaatgcag aagggctgcg tctccccaga cccacagaag ggcaaaatgc
     1201 agttcaagat gaagaaaggt agccaacagc acacaaacac acccatgatc accgacagag
     1261 tcttcaggac tttagtttct cttttcagtt ggtcatgatt gttgtactcc agcttgtgcc
     1321 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     1381 tgtcgccctc gaacttcacc tcggcgcggg tcttgtaaaa gccgtcattc ttgaagaaga
     1441 tggtgcgctc ctggatgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     1501 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     1561 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     1621 tggcatcgcc ctcaccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     1681 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatcagcc
     1741 ctccggtacc gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     1801 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttcgaaagt ttggactgca
     1861 cgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     1921 ggtagtggta ggcgagctgc acgccaccgt cctcgatgtt gtggcggatc ccgaagttcg
     1981 ccttgatgcc gttcttctgc ttgtcggcct tgatatagac ataaatgagt gaattcagct
     2041 gtttctgagc aatcctgtag atcctggtgt acaagacaat catgatggcc acagggatgt
     2101 aaaagcttat tacagaggat gagatggcat atgtcctgct gaggctggag tcacagttgt
     2161 ctatggtctc agccagggaa gtggcatttc catcagaggg gcttgtgggt tttgccttgt
     2221 gccagctgag ctgcactggg atgaaggaga tgagtacaga caaggtccat gccacactga
     2281 tcaggatgaa ggctgccttg ggggtcatct ttctctcata ccgggcaggg ctggagatag
     2341 cccaatacct gtccacgctg atcacacaga ggttgaggat ggatgcagtg gagcacatga
     2401 tgtcaaaggc cacccagatg ttacagaagg acccaaaggg ccagaagcca gcaatctcag
     2461 ccactgcctt ccagggcatg accaggacgg ccaccaagag atctgacaca gccaaggaga
     2521 tgacaaagaa gttggtcacc ttggaccgca ggtgtcggaa cctgataacg gcagcacaga
     2581 ccagcgtgtt ccccaggagc gtggacagga tgagcagcga caggaaacag gcagtgagga
     2641 tacgaacaga gaagtccctc tccaccacca gcccagtccc gtccatggca gaggtgttca
     2701 gagtcctacc ggtggtgcca acttttttgt acaaacttgt gtcaccagtg gaacctggaa
     2761 cccagagcag cagtacccat agcaggagtg tgtctgtctc catggtggcg ctagcataac
     2821 ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt
     2881 ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg
     2941 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3001 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3061 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3121 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3181 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3241 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3301 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3361 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     3421 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3481 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3541 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     3601 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3661 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     3721 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     3781 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     3841 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     3901 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     3961 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     4021 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     4081 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4141 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4201 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4261 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4321 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4381 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4441 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4501 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4561 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4621 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4681 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4741 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4801 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4861 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4921 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4981 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5041 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5101 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5161 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5221 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5281 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5341 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5401 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5461 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5521 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5581 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5641 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5701 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5761 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5821 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5881 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5941 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6001 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6061 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6121 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6301 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6781 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcac
//
