LOCUS       sequence-417679-        6662 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses the genetically-encoded fluorescent neuropeptide Y (NPY) 
            sensor GRAB_NPY1.0 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6662)
  AUTHORS   Wang H, Qian T, Zhao Y, Zhuo Y, Wu C, Osakada T, Chen P, Chen Z, Ren
            H, Yan Y, Geng L, Fu S, Mei L, Li G, Wu L, Jiang Y, Qian W, Zhang L,
            Peng W, Xu M, Hu J, Jiang M, Chen L, Tang C, Zhu Y, Lin D, Zhou JN, 
            Li Y
  TITLE     A tool kit of highly selective and sensitive genetically encoded 
            neuropeptide sensors.
  JOURNAL   Science. 2023 Nov 17;382(6672):eabq8173. doi: 
            10.1126/science.abq8173. Epub 2023 Nov 17.
  PUBMED    37972184
REFERENCE   2  (bases 1 to 6662)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2023-11-17"; volume: "382(6672):eabq8173. doi"; pages: " 
            10.1126/science.abq8173. Epub 2023 Nov 17"
COMMENT     Sequence Label: pAAV-hSyn-GRAB_NPY1.0
FEATURES             Location/Qualifiers
     source          1..6662
                     /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"
     primer_bind     1825..1846
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1952..1973)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2213..2232)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2789..3377
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2842..2862)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3260..3271)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3409..3885
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3608..3627)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3925..4065
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3925..4054
                     /label=AAV2 ITR
     rep_origin      4140..4595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4227..4246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4437..4458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4612..4631)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4731..4753
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4791..4809)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4877..4981
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4982..5050,5051..5842)
                     /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(5200..5219)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6013..6601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6502..6521
                     /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 gtaattcaac attattttcc caggttgaaa atcattcagt
      721 ccactctaat ttctcagaga agaatgccca gcttctggct tttgaaaatg atgattgtca
      781 tctgcccttg gccatgatat ttaccttagc tcttgcttat ggagctgtga tcattcttgg
      841 tgtctctgga aacctggcct tgatcataat catcttgaaa caaaaggaga tgagaaatgt
      901 taccaacatc ctgattgtga acctttcctt ctcagacttg cttgttgcca tcacgtgtct
      961 cccctttaca tttgtctaca cattaatgga ccactgggtc ttcggtgagg cgatgtgtaa
     1021 gttgaatcct tttgtgcaat gtgtttcaat cactgtgtcc attttctctc tggttctcat
     1081 tgctgtggaa cgacatcagc tgataatcaa ccctcgaggg tggagaccaa ataatagaca
     1141 tgcttatgta ggtattgctg tgatttgggt ccttgctgtg gcttcttctt tgcctttcct
     1201 gatctaccaa gtaatgactg atgagccgtt ccaaaatgta acacttgatg cgtacaaaga
     1261 caaatacgtg tgctttgatc aatttccatc ggactctcat aggttgtctt ataccactct
     1321 cctcttggtg ctgcagtatt ttggtccact ttgttttata tttatttgct acttcaagat
     1381 atatatacgc ctaaaaagga gaaacaacat gatgccaccc agccgccggg gtccggacgc
     1441 cgtcgccgcg ccgccggggg gcaccgagcg caggcccaac ggtctgggcc ccgagcgcag
     1501 cgcgggcccg gggggcgcag aggccgaacc gctgcccacc cagctcaacg gcgcccctgg
     1561 cgagcccgcg ccggccgggc cgcgcgacac cgacgcgctg gacctggagg agggaggaaa
     1621 cgtctatatc aaggccgaca agcagaagaa cggcatcaag gcgaacttcc acatccgcca
     1681 caacatcgag gacggcggcg tgcagctcgc ctaccactac cagcagaaca cccccatcgg
     1741 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca aactttcgaa
     1801 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1861 cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatgg tgagaaaggg
     1921 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1981 ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc accgagggca agctgaccct
     2041 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2101 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2161 caagtccgcc atgcccgaag gctacatcca ggagcgcacc atcttcttca aggacgacgg
     2221 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2281 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2341 caccggagca gcagcacgct ggcgcgggcg gcagaacgaa accaaaagaa tcaatatcat
     2401 gctgctctcc attgtggtag catttgcagt ctgctggctc cctcttacca tctttaacac
     2461 tgtgtttgat tggaatcatc agatcattgc tacctgcaac cacaatctgt tattcctgct
     2521 ctgccacctc acagcaatga tgtccacttg tgtcaacccc atattttatg ggttcctgaa
     2581 caaaaacttc cagagagact tgcagttctt cttcaacttt tgtgatttcc ggtctcggga
     2641 tgatgattat gaaacaatag ccatgtccac gatgcacaca gatgtttcca aaacttcttt
     2701 gaagcaagca agcccagtcg catttaaaaa aatcaacaac aatgatgata atgaaaaaat
     2761 ctaagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     2821 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2881 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2941 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3001 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3061 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3121 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3181 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     3241 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3301 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3361 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc
     3421 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     3481 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     3541 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     3601 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     3661 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     3721 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     3781 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     3841 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     3901 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3961 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4021 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4081 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4201 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4321 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4441 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     4501 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4561 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4621 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4681 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4741 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4801 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4861 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4921 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4981 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5041 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5101 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5161 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5221 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5281 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5341 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5401 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5461 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5521 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5581 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5641 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5701 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5761 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5821 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5881 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5941 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6001 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6061 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6661 gt
//
