LOCUS       sequence-466650-        6587 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hSyn1 driven expression of the consitutively active kinase activity 
            recorder positive control Kinprola_on fused to mEGFP for neuronal 
            expression through AAV transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6587)
  AUTHORS   Sun DE, Ng SW, Zheng Y, Xie S, Schwan N, Breuer P, Hoffmann DC, 
            Michel J, Azorin DD, Boonekamp KE, Winkler F, Wick W, Boutros M, Li 
            Y, Johnsson K
  TITLE     Molecular recording of cellular protein kinase activity with 
            chemical labeling.
  JOURNAL   Nat Chem Biol. 2025 Jul 10. doi: 10.1038/s41589-025-01949-6.
  PUBMED    40640555
REFERENCE   2  (bases 1 to 6587)
  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: "Nat Chem 
            Biol. 2025 Jul 10. doi: 10.1038/s41589-025-01949-6."
COMMENT     Sequence Label: pAAV_hSyn_NES-Kinprola_on-mEGFP_WRPE-SV40
FEATURES             Location/Qualifiers
     source          1..6587
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    16..604
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(69..89)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(487..498)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(607..623)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(608..624)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    653..774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(664..683)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     718..737
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   936..1065
                     /label=AAV2 ITR
     rep_origin      complement(1302..1890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1382..1401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2061..2852,2853..2921))
                     /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     2684..2703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2922..3026)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3052..3507)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3189..3210)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3401..3420
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3579..3708
                     /label=AAV2 ITR
     promoter        3790..4237
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(5871..5873,5874..5876,5877..6587,1..3)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5916..5937)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6177..6196)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6524..6545
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tgaaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
       61 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
      121 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
      181 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
      241 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
      301 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
      361 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
      421 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
      481 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
      541 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
      601 ccgcatcgat accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac
      661 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
      721 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
      781 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
      841 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
      901 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
      961 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     1021 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     1081 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1141 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1201 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1741 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1861 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1921 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1981 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2041 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2101 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2161 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2221 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2281 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2341 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2401 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2461 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2521 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2581 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2641 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2701 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2761 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2821 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2881 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2941 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3001 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     3061 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3121 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3181 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3241 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3301 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3361 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3421 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3481 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     3541 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg
     3601 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     3661 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga
     3721 ttaacccgcc atgctactta tctacgtagc catgctctag gaagatctct gcagagggcc
     3781 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
     3841 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
     3901 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
     3961 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
     4021 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
     4081 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
     4141 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
     4201 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg aattcaagct gctagcaagg
     4261 atccacccgc caccatgctg cagaacgagc ttgctcttaa gttggctgga cttgatatta
     4321 acaagactgg aggttctttt gcccgcgaga ccttccaggc cttccgcacc accgacgtcg
     4381 gccgcaagct gatcatcgat cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg
     4441 tccgcccgct gactgaagtc gagatggacc attaccgcga gccgttcctg aatcctgttg
     4501 accgcgagcc actgtggcgc ttcccaaacg agctgccaat cgccggtgag ccagcgaaca
     4561 tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca gtcccctgtc ccgaagctgc
     4621 tgttctgggg caccccaggc gttctgatcc caccggccga agccgctcgc ctggccaaaa
     4681 gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct gaatctgctg caagaagaca
     4741 acccggacct gatcggcagc gagatcgcgc gctggctgtc tactctggag attggtggca
     4801 ccggtggctc tggcggtacc ggtggctctg gcggttctat cggtactggc tttccattcg
     4861 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     4921 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     4981 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     5041 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     5101 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     5161 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     5221 tcatccgccc tatcccgacc tgggacgaat gggctccggg ttctctgcgt cgcgccaccc
     5281 tggtggaccc tcctcctccg ccaccgccac caccgccgcc gccgccacca ccacctcctc
     5341 cgcctcctcc accaccaccg ccaccgcctc caccacctgg cggcagcgcc tgtgactatc
     5401 atttaggaaa catttcaaga ctgtcaaata agcatttcca aatactacta ggagaagacg
     5461 gtaacctttt attgaatgac atttccacta atgggacctg gttaaatggg caaaaagtcg
     5521 agaagaacag ctatcagtta ctgtctcaag gtgatgaaat aaccgttggt gtaggcgtgg
     5581 aatcagatat tttatctctg gtcattttca taaacgacaa atttaagcag tgcctcgagc
     5641 agaacaaagt tgatcgcggt ggcggtatgc ataagttttc tcaagaacag atcggcgaaa
     5701 acattgtgtg cagggtcatt tgtaccacgg gtcaaattcc catccgagat ttgtcagctg
     5761 atatttcaca agtgcttaag gaaaaacgat ccataaagaa agtttggaca tttggtagaa
     5821 acccaggttc tgccaaggag accttccagg ccttccgcag cggaggttct atggtgagca
     5881 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     5941 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     6001 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     6061 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     6121 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     6181 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     6241 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     6301 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     6361 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     6421 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     6481 cccagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     6541 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaag
//
