LOCUS       sequence-454355-        6712 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 6712)
  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 6712)
  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-CaMKII(0.3)-mVenus-FKBP-K7GEF
FEATURES             Location/Qualifiers
     source          1..6712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    75..551
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     333..352
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    583..1171
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             689..700
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1098..1118
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2504..2824)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(join(2837..3547,3548..3550,3551..3553))
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2879..2900)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3228..3247
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3487..3508
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3550..3559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3880..4009
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4071..4659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4151..4170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4830..5621,5622..5690))
                     /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     5453..5472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5691..5795)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5863..5881
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5919..5941)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6041..6060
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6077..6532)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6214..6235)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6426..6445
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(6607..6712,1..35)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(6618..6712,1..35)
                     /label=AAV2 ITR
ORIGIN
        1 agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtggtt
       61 acctacaaaa tcagaaggac agggaaggga gcagtggttc acgcctgtaa tcccagcaat
      121 ttgggaggcc aaggtgggta gatcacctga gattaggagt tggagaccag cctggccaat
      181 atggtgaaac cccgtctcta ccaaaaaaac aaaaattagc tgagcctggt catgcatgcc
      241 tggaatccca acaactcggg aggctgaggc aggagaatcg cttgaaccca ggaggcggag
      301 attgcagtga gccaagattg tgccactgca ctccagcttg gttcccaata gaccccgcag
      361 gccctacagg ttgtcttccc aacttgcccc ttgctccata ccacccccct ccaccccata
      421 atattataga aggacaccta gtcagacaaa atgatgcaac ttaattttat taggacaagg
      481 ctggtgggca ctggagtggc aacttccagg gccaggagag gcactgggga ggggtcacag
      541 ggatgccacc cgtagatctc tcgagcagcg ctcggtatcg atgcggggag gcggcccaaa
      601 gggagatccg actcgtctga gggcgaaggc gaagacgcgg aagaggccgc agagccggca
      661 gcaggccgcg ggaaggaagg tccgctggat tgagggccga agggacgtag cagaaggacg
      721 tcccgcgcag aatccaggtg gcaacatagg cgagcagcca aggaaaggac gatgatttcc
      781 ccgacaacac cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa
      841 ggcaggcggc gatgagttcc gccgtggcaa tagggagggg gaaagcgaaa gtcccggaaa
      901 ggagctgaca ggtggtggca atgccccaac cagtgggggt tgcgtcagca aacacagtgc
      961 acaccacgcc acgttgcctg acaacgggcc acaactcctc ataaagagac agcaaccagg
     1021 atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga tacaaaggca
     1081 ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata ccagtcaatc
     1141 tttcacaaat tttgtaatcc agaggttgat tatcgataag cttgatatcg aattcttact
     1201 aaacgtaagt ggagaaagga tctccaggtc ccaggtgcca ccgaggaaca aggttgccat
     1261 ccttgtcact ctccactgtg ttctgagagg tcctggaggc aatggatatg gtatcgggct
     1321 ggctgctccc gtccccctgg ctgtcaccat cctctacccc atccctctta ctattgttcc
     1381 tcagtttggc tggtgttttg gggagctgga ttggctcctt taaagctcct ttaagatgaa
     1441 tgatcctttc ctgaatcact tcccgaatgt tcttgatcca ttcctgcttg gtttcgatgt
     1501 tggaagcttt gagcactgtt ttattgtctg aggatggggt gcgcccagac cacaaggcga
     1561 atttgcaggg gtcaccttct acgtgctcgg tcacacccag ctctgaggtc agtagcttgt
     1621 tcttgtaaac atatttcgtg tgtcctgaag agtctttgat ctctttgcta aacaccaagg
     1681 agatctcaaa gaggaacaga tgccgttccc gccccttccg gatcagcgac tttgggtccc
     1741 acacttgaaa ggcatcctgg agaatcaact ccccctgtac atccaagttc tcatcgaacc
     1801 cttcaagcat gctgacatgc atggcatcat tggctttctt ggggacactg agcatcacct
     1861 ccaggccatc cttgagctcc cctttccctt cttcacagca agttaaaagt tccttcagga
     1921 gcagctggta cttcgtaact ctctggacag gcttaattaa gtaggaagag atggagttgg
     1981 ccagcccgtg ccgctgctgg atctcatcaa agaaggtgcc tgcgtgctcc aggatcagct
     2041 ggttggaatc aggcttgttt ttacagtagg tgacatacat ctgaaatttg tctgcccagg
     2101 taacaaagca gtgtcccaca tcctcaggca gctgctcgta cttctccagc tctttgagga
     2161 agatgttgtt gtggaagtca tagatctcct ggatgttgcc gaagatgatg tgctccttat
     2221 tgaggatccc aggcggtatc tcttccacac cactggtcat ttcccacagg taggtctcca
     2281 agcactcgtg taggtctctt acataagcct tctcggtctg gagcagttca gccataatga
     2341 attctttctt tcgagctgac ttccgtttct cttcattgac ctcgtggtta gcatcccgga
     2401 gcttgacctc ccggtctgaa aggcttgctg ggattaggat cccacctcca gcactgccac
     2461 ctgcacttcc gccagcacta ccaccagcac taccgccagc actttccagt tttagaagct
     2521 ccacatcgaa gacgagagtg gcatgtggtg ggatgatgcc tgggtgccca gtggcaccat
     2581 aggcataatc tggagatata gtcagtttgg ctctctgacc cacactcatc tgggcaaccc
     2641 cttcttccca gcctcggatc acctcctgct tgcctagcat aaacttaaag ggcttgtttc
     2701 tgtcccggga ggaatcaaat ttctttccat cttcaagcat cccggtgtag tgcaccacgc
     2761 aggtctggcc gcgcttgggg aaggtgcgcc cgtctcctgg ggagatggtt tccacctgca
     2821 ctccaccacc agcactcttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     2881 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     2941 ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     3001 gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt
     3061 tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt
     3121 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     3181 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     3241 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     3301 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcagg ccgtagccca
     3361 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atcagcttca
     3421 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     3481 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     3541 ccttgctcac catggtggcg accggtagcg ctagccccgc tgcccccaga actaggggcc
     3601 actcgcctgc ccgtgctcct gagtgcaaac ggagaaccgg cttgactgac gagcttgggg
     3661 cttctgagca gggcactgtg gctgctcaca gcctcgcgag ccttcgtcag catccaggtc
     3721 cccatggcaa ccacctccga aacgccctgt tcccgttgcc ccggtaactg cctccccaac
     3781 acctgcctgc ctttcacttt aaaacctgct cctctttgcc ctggccttat atacacaata
     3841 ctgcaggact gtgtgggccc agaagacgcg tgcggccgca ggaaccccta gtgatggagt
     3901 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3961 gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca
     4021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4501 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5821 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5881 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5941 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6001 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6061 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     6121 tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca
     6181 aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta
     6241 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     6301 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     6361 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     6421 aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg
     6481 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     6541 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6601 ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     6661 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc ag
//
