LOCUS       sequence-410348-        6677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent vasoactive intestinal 
            peptide (VIP) sensor GRAB_VIP1.0 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6677)
  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 6677)
  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: "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_VIP1.0
FEATURES             Location/Qualifiers
     source          1..6677
                     /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     1879..1900
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2006..2027)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2267..2286)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2804..3392
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2857..2877)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3275..3286)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3424..3900
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3623..3642)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3940..4080
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3940..4069
                     /label=AAV2 ITR
     rep_origin      4155..4610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4242..4261)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4452..4473
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4627..4646)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4746..4768
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4806..4824)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4892..4996
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4997..5065,5066..5857)
                     /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(5215..5234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6028..6616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6517..6536
                     /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 gtcggacgct gctgcctccc gcgctgctga cctgctggct
      721 gctcgccccc gtgaacagca ttcacccaga atgccgattt catctggaaa tacaggagga
      781 agaaacaaaa tgtgcagagc ttctgaggtc tcaaacagaa aaacacaaag cctgcagtgg
      841 cgtctgggac aacatcacgt gctggcggcc tgccaatgtg ggagagaccg tcacggtgcc
      901 ctgcccaaaa gtcttcagca atttttacag caaagcagga aacataagca aaaactgtac
      961 gagtgacgga tggtcagaga cgttcccaga tttcgtcgat gcctgtggct acagcgaccc
     1021 ggaggatgag agcaagatca cgttttatat tctggtgaag gccatttata ccctgggcta
     1081 cagtgtctct ctgatgtctc ttgcaacagg aagcataatt ctgtgcctct tcaggaagct
     1141 gcactgcacc aggaattaca tccacctgaa cctgttcctg tccttcatcc tgagagccat
     1201 ctcagtgctg gtcaaggacg acgttctcta ctccagctct ggcacgttgc actgccctga
     1261 ccagccatcc tcctgggtgg gctgcaagct gagcctggtc ttcctgcagt actgcatcat
     1321 ggccaacttc ttctggctgc tggtggaggg gctctacctc cacaccctcc tggtggccat
     1381 gctcccccct agaaggtgct tcctggccta cctcctgatc ggatggggcc tccccaccgt
     1441 ctgcatcggt gcatggactg cggccaggct ctacttagaa gacaccggtt gctgggatac
     1501 aaacgaccac agtgtgccct ggtgggtcat acgaataccg attttaattt ccatcatcgt
     1561 caattttgtc cttttcatta gtattatacg aattttgctg cagaagttaa catccccacc
     1621 cgcgccggcc gggccgcgcg acaccgacgc gctggacctg gaggagggag gaaacgtcta
     1681 tatcaaggcc gacaagcaga agaacggcat caaggcgaac ttccacatcc gccacaacat
     1741 cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca tcggcgacgg
     1801 ccccgtgctg ctgcccgaca accactacct gagcgtgcag tccaaacttt cgaaagaccc
     1861 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1921 cggcatggac gagctgtaca agggcggtac cggagggagc atggtgagaa agggcgagga
     1981 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2041 gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga ccctgaagtt
     2101 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2161 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2221 cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2281 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2341 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacaccgg
     2401 agcagcagca cgctggcgcg ggtctcagta caagaggctg gccaagtcca cgctcctgct
     2461 tatcccgctg ttcggcgtcc actacatggt gtttgccgtg tttcccatca gcatctcctc
     2521 caaataccag atactgtttg agctgtgcct cgggtcgttc cagggcctgg tggtggccgt
     2581 cctctactgt ttcctgaaca gtgaggtgca gtgcgagctg aagcgaaaat ggcgaagccg
     2641 gtgcccgacc ccgtccgcga gccgggatta cagggtctgc ggttcctcct tctcccgcaa
     2701 cggctcggag ggcgccctgc agttccaccg cggctcccgc gcccagtcct tcctgcaaac
     2761 ggagacctcg gtcatctagg aattcgatat caagcttatc gataatcaac ctctggatta
     2821 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2881 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2941 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3001 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3061 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3121 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3181 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg
     3241 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3301 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3361 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc
     3421 tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     3481 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     3541 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     3601 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     3661 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     3721 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     3781 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     3841 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     3901 ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     3961 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4021 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4081 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4141 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4201 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4261 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4321 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4381 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4441 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4501 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4561 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4621 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4681 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4741 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4801 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4861 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4921 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4981 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5041 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5101 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5161 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5221 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5281 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5341 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5401 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5461 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5521 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5581 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5641 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5701 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5761 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5821 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5881 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5941 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6001 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6661 gccttttgct cacatgt
//
