LOCUS       sequence-411116-        6608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent 
            corticotropin-releasing factor (CRF) sensor GRAB_CRF0.5 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6608)
  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 6608)
  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_CRF0.5
FEATURES             Location/Qualifiers
     source          1..6608
                     /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     1834..1855
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1961..1982)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2222..2241)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2735..3323
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2788..2808)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3206..3217)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3355..3831
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3554..3573)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3871..4011
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3871..4000
                     /label=AAV2 ITR
     rep_origin      4086..4541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4173..4192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4383..4404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4558..4577)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4677..4699
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4737..4755)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4823..4927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4928..4996,4997..5788)
                     /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(5146..5165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5959..6547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6448..6467
                     /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 gtatgggagg gcacccgcag ctccgtctcg tcaaggccct
      721 tctccttctg gggctgaacc ccgtctctgc ctccctccag gaccagcact gcgagagcct
      781 gtccctggcc agcaacatct caggactgca gtgcaacgca tccgtggacc tcattggcac
      841 ctgctggccc cgcagccctg cggggcagct agtggttcgg ccctgccctg cctttttcta
      901 tggtgtccgc tacaatacca caaacaatgg ctaccgggag tgcctggcca atggcagctg
      961 ggccgcccgc gtgaattact ccgagtgcca ggagatcctc aatgaggaga aaaaaagcaa
     1021 ggtgcactac catgtcgcag tcatcatcaa ctacctgggc cactgtatct ccctggtggc
     1081 cctcctggtg gcctttgtcc tctttctgcg gctcaggagc atccggtgcc tgcgaaacat
     1141 catccactgg aacctcatct ccgccttcat cctgcgcaac gccacctggt tcgtggtcca
     1201 gctaaccatg agccccgagg tccaccagag caacgtgggc tggtgcaggt tggtgacagc
     1261 cgcctacaac tacttccatg tgaccaactt cttctggatg ttcggcgagg gctgctacct
     1321 gcacacagcc atcgtgctca cctactccac tgaccggctg cgcaaatgga tgttcatctg
     1381 cattggctgg ggtgtgccct tccccatcat tgtggcctgg gccattggga agctgtacta
     1441 cgacaatgag aagtgctggt ttggcaaaag gcctggggtg tacaccgact acatctacca
     1501 gggccccatg atcctggtcc tgctgatcaa tttcatcttc cttttcaaca tcgtccgcat
     1561 cctcatgacc aagctccggg catccacccc gcgcgacacc gacgcgctgg acctggagga
     1621 gggaggaaac gtctatatca aggccgacaa gcagaagaac ggcatcaagg cgaacttcca
     1681 catccgccac aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac
     1741 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa
     1801 actttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1861 cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag ggagcatggt
     1921 gagaaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1981 cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca ccgagggcaa
     2041 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2101 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2161 cgacttcttc aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa
     2221 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2281 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2341 ggagtacaac accggagcag cagcacgctg gcgcgggcgg cagtacagga aggctgtgaa
     2401 agccactctg gtgctgctgc ccctcctggg catcacctac atgctgttct tcgtcaatcc
     2461 cggggaggat gaggtctccc gggtcgtctt catctacttc aactccttcc tggaatcctt
     2521 ccagggcttc tttgtgtctg tgttctactg tttcctcaat agtgaggtcc gttctgccat
     2581 ccggaagagg tggcaccggt ggcaggacaa gcactcgatc cgtgcccgag tggcccgtgc
     2641 catgtccatc cccacctccc caacccgtgt cagctttcac agcatcaagc agtccacagc
     2701 agtcttgtaa gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     2761 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2821 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2881 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2941 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3001 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3061 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3121 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     3181 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3241 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3301 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     3361 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     3421 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     3481 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     3541 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     3601 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     3661 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     3721 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     3781 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     3841 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     3901 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3961 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     4021 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     4081 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4141 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     4201 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4261 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     4321 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4381 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     4441 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     4501 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     4561 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     4621 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4681 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     4741 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4801 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4861 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4921 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4981 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5041 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5101 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5161 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5221 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5281 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5341 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5401 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5461 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5521 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5581 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5641 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5701 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5761 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5821 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5881 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5941 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6001 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6061 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6601 tcacatgt
//
