LOCUS       sequence-412160-        6713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent somatostatin (SST) 
            sensor GRAB_SST1.0 in a cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6713)
  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 6713)
  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_SST1.0
FEATURES             Location/Qualifiers
     source          1..6713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     71..90
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   232..361
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(836..869)
                     /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(920..953)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1475..1494
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1734..1755
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1861..1882)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2829..2853)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(2857..2916)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2913..2922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2929..2962
                     /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    3013..3046
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3071..3659
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3124..3144)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3542..3553)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3691..4167
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3890..3909)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4207..4347
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4207..4336
                     /label=AAV2 ITR
     rep_origin      4422..4877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4509..4528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4719..4740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4894..4913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5013..5035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5073..5091)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5159..5263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5264..5332,5333..6124)
                     /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(5482..5501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6295..6713,1..170)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
       61 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
      241 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
      301 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      361 tgcggccgca cgcgtatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca
      421 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg
      481 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct
      541 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca
      601 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc
      661 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg
      721 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg
      781 tcgtgcctga gagcgcagtc gagaaaccgg tgccacctct agagtcgact ccggaataac
      841 ttcgtatagg atactttata cgaagttatg cagaatggta gctggattgt agctgctatt
      901 agcaatatga aacctcttaa taacttcgta tagcatacat tatacgaagt tatggcgcat
      961 atgtcattac agcttgctgg tctgcataag cccgttggct gcggcgcggt gcgcgggcgg
     1021 cgtggcctcc tgctgctgcc ggatcctgtc tggacgcggc tccgtggcgt cagcgtcctt
     1081 ggcaccagag cccttgcgga ggcacagaac cttctggaag ctctggcgga agttgtcaga
     1141 gaggaagccg tagaggacgg ggttggcaca gctgttggcg taggagagga tgaccacgaa
     1201 gaagtagagg ccggcggagg cgggctcctg gggcagcgcc acggccaggt tgacgatgtt
     1261 gacggtgaag aagggcagcc aacatcccgc aaacaccagc accaccacca acaccatcca
     1321 cgtcaccttg agctgccgcc cgcgccagcg tgctgctgct ccggtgttgt actccagctt
     1381 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1441 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1501 gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1561 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1621 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     1681 ctcggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1741 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct ttctcaccat
     1801 gctccctccg gtaccgccct tgtacagctc gtccatgcca atagtgatcc cggcggcggt
     1861 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga
     1921 ctgcacgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1981 ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc ggatgtggaa
     2041 gttcgccttg atgccgttct tctgcttgtc ggccttgata tagacgtttc ctccctcctc
     2101 caggtccagc gcgcagccca cgcgcacgcc cgccgccctc accttcacca cgatgagcag
     2161 gtagcacagg cagatgacca gcagcggcgc gaagaagccc agcacggccg tgtagatgat
     2221 gaagacggcg ccccacagcc ccacgggctc cggccagctg gcgttgcagg taccgccctc
     2281 ctgcacgtcc gcgaacacca ggagcggcag cgacatgcac agagacagga cccaggccgc
     2341 ggcgctcgcc agcttggcca cacgcgggcg gcgccagcgg gccgagctca gcgggtgcac
     2401 cactgccagg tagcggtcca cgctcatgac tgtcaggcag aagacactgg tgaactggtt
     2461 gacgccgtcc agcgtcatga ccaggcggca caggacgggg ccgaagggcc agaaggacgc
     2521 ggcgttctgc gtggccagga aaggcagccc cagcatgtac aggacgtcgg ccactgccag
     2581 gttgagaatg tagatgttgg tgacggtctt catcttggcg aagcgcagca ccacgtagat
     2641 gaccagcgtg ttcccgccca gcccggccgc acacaccagc aggtacagca cgggcaccag
     2701 caccgcccgg gcccctgccg agggcgccgg ccccaccagc gtcctgttgt caccgcctcc
     2761 agaggcagcc cccggggagg aggcgttcca gctgggcgtg gaggctggga acaggggctc
     2821 accggtggtg ccaacttttt tgtacaaact tgtgtcacca gtggaacctg gaacccagag
     2881 cagcagtacc catagcagga gtgtgtctgt ctccatggtg gcgctagcat aacttcgtat
     2941 aaagtatcct atacgaagtt atttgcctta acccagaaat tatcactgtt attctttaga
     3001 atggtgcaaa gaataacttc gtataatgta tgctatacga agttatgaat tcgatatcaa
     3061 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3121 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3181 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3241 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3301 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3361 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3421 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3481 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3541 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3601 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3661 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     3721 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3781 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3841 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3901 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3961 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     4021 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     4081 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     4141 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     4201 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4261 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4321 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     4381 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4441 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     4501 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4561 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4621 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4681 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4741 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4801 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4861 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4921 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4981 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5041 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     5101 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5281 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5341 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5401 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5461 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5521 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5581 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5641 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5701 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5761 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5821 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5881 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5941 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6001 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6061 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6121 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6181 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6241 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6301 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6361 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6421 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aag
//
