LOCUS       sequence-486784-        6827 bp    DNA     circular SYN 19-SEP-2025
DEFINITION  Expresses the chemigenetic dopamine (DA) control sensor 
            GRAB_HaloDAmut in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6827)
  AUTHORS   Zheng Y, Cai R, Wang K, Zhang J, Zhuo Y, Dong H, Zhang Y, Wang Y, 
            Deng F, Ji E, Cui Y, Fang S, Zhang X, Zhang K, Wang J, Li G, Miao X,
            Wang Z, Yang Y, Li S, Grimm J, Johnsson K, Schreiter E, Lavis L, 
            Chen Z, Mu Y, Li Y
  TITLE     In vivo multiplex imaging of dynamic neurochemical networks with 
            designed far-red dopamine sensors.
  JOURNAL   Science. 2025 Jun 5;388: 6751. doi: 
            https://doi.org/10.1126/science.adt7705.
  PUBMED    39763912
REFERENCE   2  (bases 1 to 6827)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2025-06-5"; volume: "388: 6751. doi: https"; pages: 
            "//doi.org/10.1126/science.adt7705"
COMMENT     Sequence Label: pAAV-hSyn-GRAB-HaloDAmut
FEATURES             Location/Qualifiers
     source          1..6827
                     /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..649
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     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"
     misc_feature    2954..3542
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3007..3027)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3425..3436)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3574..4050
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3773..3792)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4090..4230
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4090..4219
                     /label=AAV2 ITR
     rep_origin      4305..4760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4392..4411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4602..4623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4777..4796)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4896..4918
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4956..4974)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5042..5146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5147..5215,5216..6007)
                     /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(5365..5384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6178..6766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6667..6686
                     /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 gtaggactct gaacacctct gccatggacg ggactgggct
      721 ggtggtggag agggacttct ctgttcgtat cctcactgcc tgtttcctgt cgctgctcat
      781 cctgtccacg ctcctgggga acacgctggt ctgtgctgcc gttatcaggt tccgacacct
      841 gcggtccaag gtgaccaact tctttgtcat ctccttggct gtgtcagatc tcttggtggc
      901 cgtcctggtc atgccctgga aggcagtggc tgagattgct ggcttctggc cctttgggtc
      961 cttctgtaac atctgggtgg cctttgacat catgtgctcc actgcatcca tcctcaacct
     1021 ctgtgtgatc agcgtggaca ggtattgggc tatctccagc cctttcaggt atgagagaaa
     1081 gatgaccccc aaggcagcct tcatcctgat cagtgtggca tggaccttgt ctgtactcat
     1141 ctccttcatc ccagtgcagc tcagctggca caaggcaaaa cccacaagcc cctctgatgg
     1201 aaatgccact tccctggctg agaccataga caactgtgac tccagcctca gcaggacata
     1261 tgccatctca gcctctgtaa taccctttta catccctgtg gccatcatga ttgtctgcta
     1321 caccaggatc tacaggattg ccaagcgtcg cacccgcgtg ccacccagcc gccggggtcc
     1381 ggacgccgtc gccgcgccgc cggggggcac cgagcgcagg cccaacggta tgtacttgcc
     1441 agagatgtgc cgtgagacct tccaggccta ccgcaccacc gacgtcggcc gcaagctgat
     1501 catcaccaag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac
     1561 tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc gcgagccact
     1621 gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt
     1681 cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt tctggggcac
     1741 cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg
     1801 caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc cggacctgat
     1861 cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt tccggcggag gaacaggtgg
     1921 ttctggtgga acagggggta gcggaggtac aggaggaagt atggcggaga tcggaactgg
     1981 attcccgttt gatccgcatt atgtggaagt tctgggagag cgcatgcatt atgtggacgt
     2041 tggtcctcgt gatgggacac cagtgctgtt ccttcacggc aatccgacat cgtcgtacgt
     2101 gtggcgtaat atcatcccgc acgttgcccc cacgcaccgc tgcattgccc ctgacttaat
     2161 tggtatgggg aaaagtgata agcctgatct ggggtacttc tttgacgacc acgtacgctt
     2221 catggatgct tttattgaag cattgggttt ggaggaagta gttttggtga tccatgattg
     2281 gggtagtgct ctggggttcc attgggccaa gcgtaaccca gaacgcgtga aaggaattgc
     2341 ctttatggag ttcatccgtc cgattccaac atgggacgac tggcctgagt ttgcaaactg
     2401 gaagactctg tcggtgatca tgggtgtgtt tgtgtgctgt tggctacctt tcttcatctt
     2461 gaactgcatt cggcccttct gtgggtctgg gtggacgcag cccttctgca ttgattccaa
     2521 cacctttgac gtgtttgtgt ggtttgggtg ggctaattca tccttgaacc ccatcattta
     2581 tgcctttaat gctgattttc ggaaggcatt ttcaaccctc ttaggatgct acagactttg
     2641 ccctgcgacg aataatgcca tagagacggt gagtatcaat aacaatgggg ccgcgatgtt
     2701 ttccagccat catgagccac gaggctccat ctccaaggag tgcaatctgg tttacctgat
     2761 cccacatgct gtgggctcct ctgaggacct gaaaaaggag gaggcagctg gcatcgccag
     2821 acccttggag aagctgtccc cagccctatc ggtcatattg gactatgaca ctgacgtctc
     2881 tctggagaag atccaaccca tcacacaaaa cggtcagcac ccaacctgag aattcgatat
     2941 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3001 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3061 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3121 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3181 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3241 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3301 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3361 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3421 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3481 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3541 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     3601 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     3661 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     3721 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     3781 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     3841 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     3901 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     3961 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     4021 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     4081 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4141 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4201 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4261 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4321 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4381 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4441 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4501 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4561 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4621 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4681 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4741 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     4801 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4861 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4921 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4981 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5041 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5101 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5161 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5221 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5281 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5341 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5401 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5461 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5521 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5581 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5641 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5701 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5761 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5821 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5881 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5941 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6001 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6061 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6121 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6181 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6241 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6301 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6361 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6421 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6481 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6541 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6601 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6661 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6721 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6781 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
