LOCUS       sequence-366070-        6462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent oxytocin(OT) sensor 
            GRAB_OT1.0 in a cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6462)
  AUTHORS   Qian T, Wang H, Wang P, Geng L, Mei L, Osakada T, Wang L, Tang Y, 
            Kania A, Grinevich V, Stoop R, Lin D, Luo M, Li Y
  TITLE     A genetically encoded sensor measures temporal oxytocin release from
            different neuronal compartments.
  JOURNAL   Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01561-2.
  PUBMED    36593404
REFERENCE   2  (bases 1 to 6462)
  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: "Nat 
            Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01561-2."
COMMENT     Sequence Label: pAAV-EFS-DIO-GRAB_OTmut
FEATURES             Location/Qualifiers
     source          1..6462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        152..363
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    complement(382..415)
                     /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(466..499)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1015..1034
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1274..1295
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1401..1422)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2348..2372)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(2376..2435)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2432..2441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2448..2481
                     /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    2532..2565
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2590..3178
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2643..2663)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3061..3072)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3210..3686
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3409..3428)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3726..3866
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3726..3855
                     /label=AAV2 ITR
     rep_origin      3941..4396
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4028..4047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4238..4259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4413..4432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4532..4554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4592..4610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4678..4782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4783..4851,4852..5643)
                     /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(5001..5020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5814..6402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6303..6322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc tgcggccgca cgcgtaagct tgggcagagc gcacatcgcc cacagtcccc
      181 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
      241 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
      301 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
      361 cagtctagag tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga
      421 atggtagctg gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc
      481 atacattata cgaagttatg gcgcatatgt cacagcgtgg atggctgcga gcatcttctc
      541 tggctggagc tgtactggct caggacaaag gtagacgagt tgctcttttt gctgacgctt
      601 gtctccccag gccggctgcc tttcaggcgg cggaatgagc agcagaggaa gcgctgcaca
      661 agttcttgga agaggtggcc cgtgaagagc atgtagatcc aggggttgca gcagctgttg
      721 aggctggcca ggagcatggc gatgatgaaa ggtgaggctt ccttgggcgc atcggcatcc
      781 cagacactcc acatctgcac gaaaaagaat ggcgtccagc acacgatgaa ggccagcacg
      841 acgatgaagg tcatcttgac cgtgcggatc ttggccttag agatgagctt tgctgctgct
      901 ccggtgttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
      961 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1021 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga tgtagccttc gggcatggcg
     1081 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg
     1141 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     1201 aacttcaggg tcagcttgcc ctcggtggca tcgccctcac cctcgccgga cacgctgaac
     1261 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1321 tcctcgccct ttctcaccat gctccctccg gtaccgccct tgtacagctc gtccatgccg
     1381 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     1441 gggtctttcg aaagtttgga ctgcacgctc aggtagtggt tgtcgggcag cagcacgggg
     1501 ccgtcgccga tgggggtgtt ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg
     1561 atgttgtggc ggatgtggaa gttcgccttg atgccgttct tctgcttgtc ggccttgata
     1621 tagacgtttc ctcccgccgt cttgagccgt aaattctgcc agatcttgaa gctgataagg
     1681 ccatagcagg tggcaagcac gatgacgggc acaatgtaga cggcgagcgt gatccacgtg
     1741 atgtaggcct tgggccccca gggttgaatg aaaacggccc agcagtcgaa gacaccgtcg
     1801 gccacctcgc gcagcgagaa gatgtgcacc tgcggcgcgc tggccaccag gcagccgagc
     1861 catgtgacga gtaccgccag gcggtcggtg cggcggctca gcgagcgcag cggctggcag
     1921 atggccaggc agcggtcgag cgacatgagc agcagcaggt aggtggacgc gaacatgccc
     1981 acaacctgca ggtacttgac gaggcggcac agcaggtcgg gcccgtagaa gcggaacgtg
     2041 atgtcccaca gaagctgcgg cagcacctgg aacaccgcca ctaccaggtc ggctatgctc
     2101 aggtgcttca tgaagaagaa gaggcgcgag tgcttgtggc gcgtggtgcg cagcgctagc
     2161 agcacgcacg cgttgccgct cagcgccagg aacaggatga ggcacagcac ggccacctcc
     2221 acccgcgcca gggcctcgtt gcgctgtggc ggcccggcag tgcgattgcc ctcggttccc
     2281 ggcggcgccg cgctcccgtt gaccgcctca gcgctccagt tagccgcaaa cgcaccctca
     2341 ccggtggtgc caactttttt gtacaaactt gtgtcaccag tggaacctgg aacccagagc
     2401 agcagtaccc atagcaggag tgtgtctgtc tccatggtgg cgctagcata acttcgtata
     2461 aagtatccta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     2521 tggtgcaaag aataacttcg tataatgtat gctatacgaa gttatgaatt cgatatcaag
     2581 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2641 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2701 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2761 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2821 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2881 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2941 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3001 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3061 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3121 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3181 cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
     3241 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     3301 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     3361 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     3421 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     3481 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     3541 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     3601 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     3661 gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg
     3721 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3781 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     3841 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     3901 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     3961 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     4021 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4081 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4141 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4201 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4261 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4321 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4381 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4441 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4501 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4561 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4621 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4681 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4741 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4801 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4861 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4921 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4981 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5041 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5101 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5161 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5221 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5281 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5341 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5401 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5461 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5521 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5581 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5641 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5701 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5761 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5821 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5881 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5941 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6001 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6061 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6121 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6181 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6241 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6301 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6361 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6421 tttttacggt tcctggcctt ttgctggcct tttgctcaca tg
//
