LOCUS       sequence-412589-        6953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent vasoactive intestinal 
            peptide (VIP) sensor GRAB_VIP1.0 in a cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6953)
  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 6953)
  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-DIO-GRAB_VIP1.0
FEATURES             Location/Qualifiers
     source          1..6953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   119..259
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   119..248
                     /label=AAV2 ITR
     rep_origin      334..789
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(421..440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     631..652
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(806..825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     925..947
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(985..1003)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1071..1175
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1176..1244,1245..2036)
                     /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(1394..1413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2207..2795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2696..2715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2857..2986
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(3461..3494)
                     /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(3545..3578)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     4082..4101
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4341..4362
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4468..4489)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5694..5718)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(5722..5781)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      5778..5787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    5794..5827
                     /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    5878..5911
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5936..6524
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5989..6009)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6407..6418)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(6556..6953,1..79)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6755..6774)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
       61 tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag
      121 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
      181 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
      241 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      301 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
      361 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
      421 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      481 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      541 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      601 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      661 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
      721 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
      781 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
      841 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      901 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      961 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     1021 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1081 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1141 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1201 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1261 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1321 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1381 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1441 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1501 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1561 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1621 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1681 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1741 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1801 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1861 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1921 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1981 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2041 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2101 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2161 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2221 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2281 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2341 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2401 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2461 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2521 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2581 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2641 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2701 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2761 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2821 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     2881 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     2941 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
     3001 atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca
     3061 acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg
     3121 cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc
     3181 ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg
     3241 caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg
     3301 caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc
     3361 gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg
     3421 cagtcgagaa accggtgcca cctctagagt cgactccgga ataacttcgt ataggatact
     3481 ttatacgaag ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct
     3541 cttaataact tcgtatagca tacattatac gaagttatgg cgcatatgct agatgaccga
     3601 ggtctccgtt tgcaggaagg actgggcgcg ggagccgcgg tggaactgca gggcgccctc
     3661 cgagccgttg cgggagaagg aggaaccgca gaccctgtaa tcccggctcg cggacggggt
     3721 cgggcaccgg cttcgccatt ttcgcttcag ctcgcactgc acctcactgt tcaggaaaca
     3781 gtagaggacg gccaccacca ggccctggaa cgacccgagg cacagctcaa acagtatctg
     3841 gtatttggag gagatgctga tgggaaacac ggcaaacacc atgtagtgga cgccgaacag
     3901 cgggataagc aggagcgtgg acttggccag cctcttgtac tgagacccgc gccagcgtgc
     3961 tgctgctccg gtgttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     4021 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     4081 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggatgt agccttcggg
     4141 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
     4201 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     4261 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcaccct cgccggacac
     4321 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     4381 gaacagctcc tcgccctttc tcaccatgct ccctccggta ccgcccttgt acagctcgtc
     4441 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     4501 ctcgttgggg tctttcgaaa gtttggactg cacgctcagg tagtggttgt cgggcagcag
     4561 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg taggcgagct gcacgccgcc
     4621 gtcctcgatg ttgtggcgga tgtggaagtt cgccttgatg ccgttcttct gcttgtcggc
     4681 cttgatatag acgtttcctc cctcctccag gtccagcgcg tcggtgtcgc gcggcccggc
     4741 cggcgcgggt ggggatgtta acttctgcag caaaattcgt ataatactaa tgaaaaggac
     4801 aaaattgacg atgatggaaa ttaaaatcgg tattcgtatg acccaccagg gcacactgtg
     4861 gtcgtttgta tcccagcaac cggtgtcttc taagtagagc ctggccgcag tccatgcacc
     4921 gatgcagacg gtggggaggc cccatccgat caggaggtag gccaggaagc accttctagg
     4981 ggggagcatg gccaccagga gggtgtggag gtagagcccc tccaccagca gccagaagaa
     5041 gttggccatg atgcagtact gcaggaagac caggctcagc ttgcagccca cccaggagga
     5101 tggctggtca gggcagtgca acgtgccaga gctggagtag agaacgtcgt ccttgaccag
     5161 cactgagatg gctctcagga tgaaggacag gaacaggttc aggtggatgt aattcctggt
     5221 gcagtgcagc ttcctgaaga ggcacagaat tatgcttcct gttgcaagag acatcagaga
     5281 gacactgtag cccagggtat aaatggcctt caccagaata taaaacgtga tcttgctctc
     5341 atcctccggg tcgctgtagc cacaggcatc gacgaaatct gggaacgtct ctgaccatcc
     5401 gtcactcgta cagtttttgc ttatgtttcc tgctttgctg taaaaattgc tgaagacttt
     5461 tgggcagggc accgtgacgg tctctcccac attggcaggc cgccagcacg tgatgttgtc
     5521 ccagacgcca ctgcaggctt tgtgtttttc tgtttgagac ctcagaagct ctgcacattt
     5581 tgtttcttcc tcctgtattt ccagatgaaa tcggcattct gggtgaatgc tgttcacggg
     5641 ggcgagcagc cagcaggtca gcagcgcggg aggcagcagc gtccgaccgg tggtgccaac
     5701 ttttttgtac aaacttgtgt caccagtgga acctggaacc cagagcagca gtacccatag
     5761 caggagtgtg tctgtctcca tggtggcgct agcataactt cgtataaagt atcctatacg
     5821 aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata
     5881 acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca
     5941 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6001 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6061 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6121 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6181 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6241 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6301 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta
     6361 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6421 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6481 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct
     6541 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     6601 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     6661 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     6721 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     6781 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     6841 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     6901 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc cta
//
