LOCUS       sequence-412156-        6887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent cholecystokinin (CCK) 
            sensor GRAB_CCK1.0 in a cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6887)
  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 6887)
  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_CCK1.0
FEATURES             Location/Qualifiers
     source          1..6887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..68
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   210..339
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(814..847)
                     /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(898..931)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1441..1460
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1700..1721
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1827..1848)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2981..3005)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(3009..3068)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3065..3074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3081..3114
                     /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    3165..3198
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3223..3811
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3276..3296)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3694..3705)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3843..4319
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4042..4061)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4359..4499
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4359..4488
                     /label=AAV2 ITR
     rep_origin      4574..5029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4661..4680)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4871..4892
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5046..5065)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5165..5187
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5225..5243)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5311..5415
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5416..5484,5485..6276)
                     /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(5634..5653)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6447..6887,1..148)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
       61 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      121 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      181 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
      241 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
      301 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgtatgaggc
      361 ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca
      421 ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg
      481 cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc
      541 gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc
      601 cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg
      661 cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag
      721 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga
      781 gaaaccggtg ccacctctag agtcgactcc ggaataactt cgtataggat actttatacg
      841 aagttatgca gaatggtagc tggattgtag ctgctattag caatatgaaa cctcttaata
      901 acttcgtata gcatacatta tacgaagtta tggcgcatat gtcagccagg gcccagtgtg
      961 ctgatggtgg tgtagctaag cctggacagc gaagcaatgg agggagtggg agggtcctca
     1021 tcgggaagag ccctggggcg agctcgtgga ggccgggggc agcagcgagc gcaagtttcc
     1081 aggcaggcct ggcgaaagcg acggtgcatg aagcagtaga ccagggggtt gacacaggcc
     1141 gaggcgtagc tcagcaagtg aatgaaggag ataggagcac ccgagagtgc tcggtgtgca
     1201 cccgggccat caaaggcgcg ccacgtgttg gcactataaa ctggcaacca acacagaaaa
     1261 aaaagcacaa cgatcaccag caacattcgc accacgcgct tcttagccag cagctttgcg
     1321 cctgctccgg tgttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     1381 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     1441 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggatgta gccttcgggc
     1501 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc
     1561 acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     1621 cagatgaact tcagggtcag cttgcccaag gtggcatcgc cctcaccctc gccgtgcacg
     1681 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     1741 aacagctcct cgccctttct caccatgctc cctccggtac cgcccttgta cagctcgtcc
     1801 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     1861 tcgttggggt ctttcgaaag tttggactgc acgctcaggt agtggttgtc gggcagcagc
     1921 acggggccgt cgccgatggg ggtgttctgc tggtagtggc tggcgagctg cacgccgccg
     1981 tcctcgatgt tgtggcggat cacgaaagtc gccttgatgc cgttcttctg cttgtcggcc
     2041 ttgatataga cgtttcctcc ctcctccagg tccagcgcgt cggtgtcgcg cggcccggcc
     2101 ggcgcgggct cgccaggggc gccgttgagc tgggtgggca gcggttcggc ctctgcgccc
     2161 cccgggcccg cgctgcgctc ggggcccaga ccgttgggcc tgcgctcggt gccccccggc
     2221 ggaaggcgaa gccctaagta gagctcgcga gagataagcc cgtaggccac ggccataacc
     2281 acacccggga tgaagaacaa gagcagaagc agcagtacgg accaggtctg gcggacccgc
     2341 gcactgggcc agcgatgcac gcactgcagc acacgaggcc ccactggttg cacgacagtg
     2401 tacacggggt agggcaccat gagtagtccg gacagcagcc acgtggctac aatcacgcga
     2461 gccgcgtggg agcgcgtctg ccacactcgt gcctggcctg gtcggcagat ggcgctgtac
     2521 cgctccagtg cgatggccac gaggcttagc gtagacacac tcacagacac ccccatgagg
     2581 taggaaaccg ccttgcagat gacggtgcca aagatccatg tgcccatgac attgggcagg
     2641 agggtgaagg gcatgcaagc cacagccagc aggaggtcgc tgactgccag tgagaggagg
     2701 aaggcattgg tgacagtcct caggcggcgg ctcagtccca ggaccacgat gatgagcata
     2761 tttcctccaa cgctcatcag gaagatcact gcgtaaagag tgattctaat ggccagctcc
     2821 aattctcgtg tcccggctcc gcgaatgcga gggggctcgc agctgaggtt gcccacactg
     2881 ctgctgttga ggagaggcgc ccccgggcgg cacagggaag cccccggccc gggtccggtt
     2941 ccctgcacgc tccggttcag ctttagcagc tcaccggtgg tgccaacttt tttgtacaaa
     3001 cttgtgtcac cagtggaacc tggaacccag agcagcagta cccatagcag gagtgtgtct
     3061 gtctccatgg tggcgctagc ataacttcgt ataaagtatc ctatacgaag ttatttgcct
     3121 taacccagaa attatcactg ttattcttta gaatggtgca aagaataact tcgtataatg
     3181 tatgctatac gaagttatga attcgatatc aagcttatcg ataatcaacc tctggattac
     3241 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3301 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3361 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3421 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3481 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3541 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3601 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     3661 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3721 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3781 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct
     3841 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     3901 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     3961 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     4021 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     4081 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     4141 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     4201 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     4261 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     4321 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     4381 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4441 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     4501 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4561 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4621 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     4681 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4741 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4801 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4861 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4921 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4981 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     5041 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5101 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5161 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5221 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5281 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5341 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5401 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5461 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5521 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5581 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5641 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5701 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5761 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5821 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5881 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5941 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6001 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6061 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6121 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6181 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6241 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6301 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6361 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6421 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6481 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6541 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6601 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6661 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6721 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6781 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6841 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatcc
//
