LOCUS       sequence-307197-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the red fluorescent medium-affinity DA sensor GRAB_rDA1m 
            in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  AUTHORS   Sun F, Zhou J, Dai B, Qian T, Zeng J, Li X, Zhuo Y, Zhang Y, Wang Y,
            Qian C, Tan K, Feng J, Dong H, Lin D, Cui G, Li Y
  TITLE     Next-generation GRAB sensors for monitoring dopaminergic activity in
            vivo.
  JOURNAL   Nat Methods. 2020 Nov;17(11):1156-1166. doi: 
            10.1038/s41592-020-00981-9. Epub 2020 Oct 21.
  PUBMED    33087905
REFERENCE   2  (bases 1 to 6224)
  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"; doi: 
            "10.1038/s41592-020-00981-9"; journalName: "Nat Methods"; date: 
            "2020-11"; volume: "17"; issue: "11"; pages: "1156-1166"
COMMENT     Sequence Label: pAAV-hsyn-GRAB_rDA1m
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     587..646
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2044..2063
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2404..2424)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2822..2833)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2971..3447
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3170..3189)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3487..3627
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3487..3616
                     /label=AAV2 ITR
     rep_origin      3702..4157
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3789..3808)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3999..4020
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4174..4193)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4293..4315
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4353..4371)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4439..4543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4544..4612,4613..5404)
                     /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(4762..4781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5575..6163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6064..6083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
      181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac
      601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
      661 caaaaaagtt ggcaccaccg gtgatccact gaatctgtcc tggtatgatg atgatctgga
      721 gaggcagaac tggagccggc ccttcaacgg gtcagacggg aaggcggaca gaccccacta
      781 caactactat gccacactgc tcaccctgct catcgctgtc atcgtcttcg gcaacgtgct
      841 ggtgtgcatg gctgtgtccc gcgagaaggc gctgcagacc accaccaact acctgatcgt
      901 cagcctcgca gtggccgacc tcctcgtcgc cacactggtc atgccctggg ttgtctacct
      961 ggaggtggta ggtgagtgga aattcagcag gattcactgt gacatcttcg tcactctgga
     1021 cgtcatgatg tgcacggcga gcatcctgaa cttgtgtgcc atcagcatcg acaggtacac
     1081 agctgtggcc atgcccatgc tgtacaatac gcgctacagc tccaagcgcc gggtcaccgt
     1141 catgatctcc atcgtctggg tcctgtcctt caccatctcc tgcccactcc tcttcggact
     1201 caataacgca gaccagaacg agtgcatcat tgccaacccg gccttcgtgg tctactcctc
     1261 catcgtctcc ttctacgtgc ccttcattgt caccctgctg gtctacatca agatctacat
     1321 tgtcctccgc agacgccgca agcgagtccc cgtgtgttcc gaggttatgt accccgagga
     1381 cggcgccctg aagagcgaga tcaagaaggg gctgaggctg aaggacggcg gccactacgc
     1441 cgccgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacat
     1501 cgtcgacatc aagttggaca tcgtgtccca caacgaggac tacaccatcg tggaacagtg
     1561 cgaacgcgcc gaggcccgcc actccaccgg cggcatggac gagctgtaca agggaggtac
     1621 aggcgggagt ctggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     1681 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     1741 gggcgagggc cgcccctacg aggcctttca gaccgctaag ctgaaggtga ccaagggtgg
     1801 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     1861 cattaagcac ccagccgaca tccccgacta cttcaagctg tccttccccg agggcttcag
     1921 gtgggagcgc gtgatgaact tcgaggacgg cggcattatt cacgttaacc aggactcctc
     1981 cctgcaggac ggctacttca tctacaaggt gaagctgcgc ggcaccaact tcccccccga
     2041 cggccccgta atgcagaaga agaccatggg ctgggagctg acggtccagc agctcgagaa
     2101 gaaagccact cagatgctcg ccattgttct cggcgtgttc atcatctgct ggctgccctt
     2161 cttcatcaca cacatcctga acatacactg tgactgcaac atcccgcctg tcctgtacag
     2221 cgccttcacg tggctgggct atgtcaacag cgccgtgaac cccatcatct acaccacctt
     2281 caacattgag ttccgcaagg ccttcctgaa gatcctccac tgctgagaat tcgatatcaa
     2341 gcttatcgat catcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2401 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2461 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2521 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2581 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2641 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2701 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2761 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2821 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2881 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2941 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     3001 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3061 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3121 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3181 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3241 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3301 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3361 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3421 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3481 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3541 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3601 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3661 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     3781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3841 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3901 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4021 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4081 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4141 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4201 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4261 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4321 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4381 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4441 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4501 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4561 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4621 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4681 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4741 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4801 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4861 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4921 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4981 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5041 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5101 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5161 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5221 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5281 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5341 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5401 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5701 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
