LOCUS       sequence-454358-        6745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A component of SYNCit-K (Synapse-targeted chemically induced 
            translocation of Kalirin-7).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6745)
  AUTHORS   Sawada T, Iino Y, Yoshida K, Okazaki H, Nomura S, Shimizu C, Arima 
            T, Juichi M, Zhou S, Kurabayashi N, Sakurai T, Yagishita S, 
            Yanagisawa M, Toyoizumi T, Kasai H, Shi S
  TITLE     Prefrontal synaptic regulation of homeostatic sleep pressure 
            revealed through synaptic chemogenetics.
  JOURNAL   Science. 2024 Sep 27;385(6716):1459-1465. doi: 
            10.1126/science.adl3043. Epub 2024 Sep 26.
  PUBMED    39325885
REFERENCE   2  (bases 1 to 6745)
  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"; doi: 
            "10.1126/science.adl3043"; journalName: "Science"; date: 
            "2024-09-27- 27"; volume: "385"; issue: "6716"; pages: "1459-1465"
COMMENT     Sequence Label: pAAV-hSyn-PSD-Delta-1,2-FRBx3
FEATURES             Location/Qualifiers
     source          1..6745
                     /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      451..460
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1915..2193
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             2224..2502
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             2533..2811
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             2818..2844
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2872..3460
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2925..2945)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3343..3354)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3492..3968
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3691..3710)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4008..4148
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4008..4137
                     /label=AAV2 ITR
     rep_origin      4223..4678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4310..4329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4520..4541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4695..4714)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4814..4836
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4874..4892)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4960..5064
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5065..5133,5134..5925)
                     /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(5283..5302)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6096..6684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6585..6604
                     /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 gcgtcttctg ggcccacaca gtcctgcagt attgtgtata
      181 taaggccagg gcaaagagga gcaggtttta aagtgaaagg caggcaggtg ttggggaggc
      241 agttaccggg gcaacgggaa cagggcgttt cggaggtggt tgccatgggg acctggatgc
      301 tgacgaaggc tcgcgaggct gtgagcagcc acagtgccct gctcagaagc cccaagctcg
      361 tcagtcaagc cggttctccg tttgcactca ggagcacggg caggcgagtg gcccctagtt
      421 ctgggggcag cggggctagc gctaccggtc gccaccatgg actgtctctg tatagtgaca
      481 accaagaaat accgctacca agatgaagac acgccccctc tggaacacag cccggcccac
      541 ctccccaacc aggccaattc tccccctgtg attgtcaaca cggacaccct agaagcccca
      601 ggatatgagt tgcaggtgaa tggaacagag ggggagatgg agtatgagcg gatcgtgatc
      661 catcggggct ccaccggcct gggcttcaac atcgtgggcg gcgaggatgg tgaaggcatc
      721 ttcatctcct tcatccttgc tgggggtcca gccgacctca gtggggagct acggaagggg
      781 gaccagatcc tgtcggtcaa tggtgttgac ctccgcaatg ccagtcacga acaggctgcc
      841 attgccctga agaatgcggg tcagacggtc acgatcatcg ctcagtataa accagaagag
      901 tatagtcgat tcgaggccaa gatccatgat cttcgggaac agctcatgaa tagtagccta
      961 ggctcaggga ctgcatcctt gcgaagcaac cccaagaggg gcttctacat tagggccctg
     1021 tttgattacg acaagaccaa ggactgcggt ttcttgagcc aggccctgag cttccgcttc
     1081 ggggatgtgc ttcatgtcat tgacgctggt gacgaagagt ggtggcaagc acggcgggtc
     1141 cactccgaca gtgagaccga cgacattggc ttcattccca gcaaacggcg ggtcgagcga
     1201 cgagagtggt caaggttaaa ggccaaggac tggggctcca gctctggatc acagggtcga
     1261 gaagactcgg ttctgagcta tgagacggtg acccagatgg aagtgcacta tgctcgtccc
     1321 atcatcatcc ttggacccac caaagaccgt gccaacgatg atcttctctc cgagttcccc
     1381 gacaagtttg gatcctgtgt ccctcatacg acacgtccta agcgggaata tgagatagac
     1441 ggccgggatt accactttgt ctcctcccgg gagaaaatgg agaaggacat ccaggcacac
     1501 aagttcattg aggctggcca gtacaacagc cacctctatg ggaccagcgt ccagtctgtg
     1561 cgagaggtag cagagcaggg gaagcactgc atcctcgatg tctcggccaa tgccgtgcgg
     1621 cggctgcagg cggcccacct gcaccccatc gccatcttca tccgtccccg ctccctggag
     1681 aatgtgctag agatcaataa gcggatcaca gaggagcaag cccggaaagc cttcgacaga
     1741 gccacgaagc tggagcagga gttcacagag tgcttctcag ccatcgtaga gggcgacagc
     1801 tttgaagaga tctatcacaa agtgaaacgt gtcattgaag acctctcagg cccctacatc
     1861 tgggtcccag cccgagagag actcgatagt gctggtagtg ctggtagtgc tggtatcctc
     1921 tggcatgaga tgtggcatga aggcctggaa gaggcatctc gtttgtactt tggggaaagg
     1981 aacgtgaaag gcatgtttga ggtgctggag cccttgcatg ctatgatgga acggggcccc
     2041 cagactctga aggaaacatc ctttaatcag gcctatggtc gagatttaat ggaggcccaa
     2101 gagtggtgca ggaagtacat gaaatcaggg aatgtcaagg acctcctcca agcctgggac
     2161 ctctattatc atgtgttccg acgaatctca aaggatagtg ctggtagtgc tggtagtgct
     2221 ggtatcctct ggcatgagat gtggcatgaa ggcctggaag aggcatctcg tttgtacttt
     2281 ggggaaagga acgtgaaagg catgtttgag gtgctggagc ccttgcatgc tatgatggaa
     2341 cggggccccc agactctgaa ggaaacatcc tttaatcagg cctatggtcg agatttaatg
     2401 gaggcccaag agtggtgcag gaagtacatg aaatcaggga atgtcaagga cctcctccaa
     2461 gcctgggacc tctattatca tgtgttccga cgaatctcaa aggatagtgc tggtagtgct
     2521 ggtagtgctg gtatcctctg gcatgagatg tggcatgaag gcctggaaga ggcatctcgt
     2581 ttgtactttg gggaaaggaa cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct
     2641 atgatggaac ggggccccca gactctgaag gaaacatcct ttaatcaggc ctatggtcga
     2701 gatttaatgg aggcccaaga gtggtgcagg aagtacatga aatcagggaa tgtcaaggac
     2761 ctcctccaag cctgggacct ctattatcat gtgttccgac gaatctcaaa gactagttac
     2821 ccatacgatg ttccagatta cgcttaagaa ttcgatatca agcttatcga taatcaacct
     2881 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2941 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3001 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3061 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3121 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3181 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3241 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     3301 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3361 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3421 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     3481 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3541 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3601 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3661 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3721 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3781 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3841 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     3901 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3961 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     4021 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4081 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4141 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4201 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4261 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4321 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4381 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4441 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4501 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4561 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4621 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4681 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4741 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4801 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4861 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4921 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4981 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5041 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5101 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5161 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5221 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5281 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5341 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5401 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5461 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5521 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5581 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5641 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5701 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5761 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5821 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5881 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5941 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6001 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6061 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6241 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6301 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6361 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6421 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6481 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6541 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6601 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6661 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6721 ttttgctggc cttttgctca catgt
//
